Structured Review

Leica Microsystems ultrathin sections
Ultrastructure of Mf. australis strain IT-1. Ultrastructure of Mf. australis strain IT-1. a Whole mount TEM image showing a single magnetosome chain, P-rich (P) and sulfur (S) granules; ( b ) <t>Ultrathin</t> section TEM image of high pressure frozen and freeze-substituted cells showing P-rich (P) and sulfur (S) granules, two magnetosomes ( black arrows ), a flagella bundle (F) associated with chemoreceptor array ( white arrows ), and a fibrillar layer at the cell surface ( arrowheads ). Uncharacterized globular structures (G) embedded in an electron-lucent material (asterisks) can be observed
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Images

1) Product Images from "Combined genomic and structural analyses of a cultured magnetotactic bacterium reveals its niche adaptation to a dynamic environment"

Article Title: Combined genomic and structural analyses of a cultured magnetotactic bacterium reveals its niche adaptation to a dynamic environment

Journal: BMC Genomics

doi: 10.1186/s12864-016-3064-9

Ultrastructure of Mf. australis strain IT-1. Ultrastructure of Mf. australis strain IT-1. a Whole mount TEM image showing a single magnetosome chain, P-rich (P) and sulfur (S) granules; ( b ) Ultrathin section TEM image of high pressure frozen and freeze-substituted cells showing P-rich (P) and sulfur (S) granules, two magnetosomes ( black arrows ), a flagella bundle (F) associated with chemoreceptor array ( white arrows ), and a fibrillar layer at the cell surface ( arrowheads ). Uncharacterized globular structures (G) embedded in an electron-lucent material (asterisks) can be observed
Figure Legend Snippet: Ultrastructure of Mf. australis strain IT-1. Ultrastructure of Mf. australis strain IT-1. a Whole mount TEM image showing a single magnetosome chain, P-rich (P) and sulfur (S) granules; ( b ) Ultrathin section TEM image of high pressure frozen and freeze-substituted cells showing P-rich (P) and sulfur (S) granules, two magnetosomes ( black arrows ), a flagella bundle (F) associated with chemoreceptor array ( white arrows ), and a fibrillar layer at the cell surface ( arrowheads ). Uncharacterized globular structures (G) embedded in an electron-lucent material (asterisks) can be observed

Techniques Used: Transmission Electron Microscopy

2) Product Images from "Coliform Bacteria for Bioremediation of Waste Hydrocarbons"

Article Title: Coliform Bacteria for Bioremediation of Waste Hydrocarbons

Journal: BioMed Research International

doi: 10.1155/2017/1838072

Transmission electron micrographs of ultrathin sections of E . coli cells from domestic sewage grown in n -hexadecane-free medium (a) and in n -hexadecane-containing medium (b) and of E . coli cells from wheat straw grown in n -hexadecane-free medium (c) and
Figure Legend Snippet: Transmission electron micrographs of ultrathin sections of E . coli cells from domestic sewage grown in n -hexadecane-free medium (a) and in n -hexadecane-containing medium (b) and of E . coli cells from wheat straw grown in n -hexadecane-free medium (c) and

Techniques Used: Transmission Assay

3) Product Images from "Fluorescent Magnetopolymersomes: A Theranostic Platform to Track Intracellular Delivery"

Article Title: Fluorescent Magnetopolymersomes: A Theranostic Platform to Track Intracellular Delivery

Journal: Materials

doi: 10.3390/ma10111303

TEM ultrathin sections of HeLa cells after 12 h incubation with fluorescent magnetopolymersomes (PBD(1200)- b -PEO(600), 10% SPION, 5% DEAC). ( A ) Overview of a typical preparation showing few polymersomes as dark spherical objects and ( B ) peripheral cell region with a rare observation of an internalized magnetopolymersome. The inset depicts a close-up of a multilamellar magnetopolymersome judged from contrast and membrane thickness.
Figure Legend Snippet: TEM ultrathin sections of HeLa cells after 12 h incubation with fluorescent magnetopolymersomes (PBD(1200)- b -PEO(600), 10% SPION, 5% DEAC). ( A ) Overview of a typical preparation showing few polymersomes as dark spherical objects and ( B ) peripheral cell region with a rare observation of an internalized magnetopolymersome. The inset depicts a close-up of a multilamellar magnetopolymersome judged from contrast and membrane thickness.

Techniques Used: Transmission Electron Microscopy, Incubation

TEM ultrathin sections of HeLa cells after 12 h incubation with cationic fluorescent magnetopolymersomes. Vesicles (50% PBD- b -PEO-COOH; 2% DEAC) containing 10% w/w SPION were formed at 2 mg/mL via solvent inversion and subsequently coated in 10× b -PEI(800) before being purified by gel permeation chromatography. Overview in ( A ) shows elevated levels of cationically modified magnetopolymersomes taken up (black spots indicated by arrows). The sequence ( B – F ) shows higher magnification images of the apparent progression of hydrolytic degradation of SPION-loaded polymersomes after internalization.
Figure Legend Snippet: TEM ultrathin sections of HeLa cells after 12 h incubation with cationic fluorescent magnetopolymersomes. Vesicles (50% PBD- b -PEO-COOH; 2% DEAC) containing 10% w/w SPION were formed at 2 mg/mL via solvent inversion and subsequently coated in 10× b -PEI(800) before being purified by gel permeation chromatography. Overview in ( A ) shows elevated levels of cationically modified magnetopolymersomes taken up (black spots indicated by arrows). The sequence ( B – F ) shows higher magnification images of the apparent progression of hydrolytic degradation of SPION-loaded polymersomes after internalization.

Techniques Used: Transmission Electron Microscopy, Incubation, Purification, GPC Assay, Modification, Sequencing

4) Product Images from "Agitation Modules: Flexible Means to Accelerate Automated Freeze Substitution"

Article Title: Agitation Modules: Flexible Means to Accelerate Automated Freeze Substitution

Journal: Journal of Histochemistry and Cytochemistry

doi: 10.1369/0022155418786698

Immunogold labeling of Lowicryl HM20 sections of HP-frozen, low-temperature embedded immature anthers of Arabidopsis thaliana generated by accelerated FS under agitation. (A) Overview image showing a middle layer cell from an anther surrounded by anti-xyloglucan labeled cell wall (cw). (B) Aggregation of intracellular vesicle-like xyloglucan-positive structures. (C) Overview image showing anti-α-tubulin labeling close to the plasma membrane. (D) Details of immunolabeled cortical microtubules (arrows) aligned almost in parallel to the section plane. Typical for postembedding labeling, the gold-conjugated antibodies are concentrated at regions where the MTs meet the surface of the ultrathin section. Scale bars = 1 µm in A and C; 300 nm in B and D. Abbreviations: n, nucleus; v, vacuole; chl, chloroplast; HP, high-pressure; FS, freeze substitution; MT, microtubule.
Figure Legend Snippet: Immunogold labeling of Lowicryl HM20 sections of HP-frozen, low-temperature embedded immature anthers of Arabidopsis thaliana generated by accelerated FS under agitation. (A) Overview image showing a middle layer cell from an anther surrounded by anti-xyloglucan labeled cell wall (cw). (B) Aggregation of intracellular vesicle-like xyloglucan-positive structures. (C) Overview image showing anti-α-tubulin labeling close to the plasma membrane. (D) Details of immunolabeled cortical microtubules (arrows) aligned almost in parallel to the section plane. Typical for postembedding labeling, the gold-conjugated antibodies are concentrated at regions where the MTs meet the surface of the ultrathin section. Scale bars = 1 µm in A and C; 300 nm in B and D. Abbreviations: n, nucleus; v, vacuole; chl, chloroplast; HP, high-pressure; FS, freeze substitution; MT, microtubule.

Techniques Used: Labeling, Generated, Immunolabeling

5) Product Images from "Extracellular Vesicles Mediate Receptor-Independent Transmission of Novel Tick-Borne Bunyavirus"

Article Title: Extracellular Vesicles Mediate Receptor-Independent Transmission of Novel Tick-Borne Bunyavirus

Journal: Journal of Virology

doi: 10.1128/JVI.02490-15

SFTS virus NSs induces the formation of endosome-like structures. Ultrastructure analyses of SFTS virus NSs-expressing cells (A) and SFTS virus-infected cells (B) show cytoplasmic structures reminiscent of early endosomes (arrows) in ultrathin sections. Immunogold staining of distinct ultrathin sections shows cytoplasmic structures positive for SFTS virus NSs and Rab5 (C) or SFTS virus NSs and LC3B (D). A solid red arrows indicate SFTS virus NSs in panels C and D, while a red triangle (in panel C) and a green triangle (in panel D) indicate the detection of Rab5 and LC3B, respectively. Ultrathin sections of mock-infected cells were also labeled, as indicated above (not shown), to ensure the specificity of antibody. Representatives images are shown.
Figure Legend Snippet: SFTS virus NSs induces the formation of endosome-like structures. Ultrastructure analyses of SFTS virus NSs-expressing cells (A) and SFTS virus-infected cells (B) show cytoplasmic structures reminiscent of early endosomes (arrows) in ultrathin sections. Immunogold staining of distinct ultrathin sections shows cytoplasmic structures positive for SFTS virus NSs and Rab5 (C) or SFTS virus NSs and LC3B (D). A solid red arrows indicate SFTS virus NSs in panels C and D, while a red triangle (in panel C) and a green triangle (in panel D) indicate the detection of Rab5 and LC3B, respectively. Ultrathin sections of mock-infected cells were also labeled, as indicated above (not shown), to ensure the specificity of antibody. Representatives images are shown.

Techniques Used: Expressing, Infection, Staining, Labeling

6) Product Images from "High Resolution Imaging of Temporal and Spatial Changes of Subcellular Ascorbate, Glutathione and H2O2 Distribution during Botrytis cinerea Infection in Arabidopsis"

Article Title: High Resolution Imaging of Temporal and Spatial Changes of Subcellular Ascorbate, Glutathione and H2O2 Distribution during Botrytis cinerea Infection in Arabidopsis

Journal: PLoS ONE

doi: 10.1371/journal.pone.0065811

Changes in the size and fine structure of chloroplasts. Size and fine structures of chloroplasts was evaluated by TEM on longitudinal ultrathin sections within the mesophyll of leaves from Arabidopsis thaliana Col-0 leaves inoculated with Botrytis cinerea and compared to CL. Measurements were performed 0 h ( = CL, represented by the dotted line), 12, 24, 48 and 96 hpi at the IS and the AIS. Shown are means with standard errors. n > 100 chloroplasts per treatment. Significant differences were calculated using the Mann-Whitney U-test; *, **, and ***, respectively, indicate significance at the 0.05, 0.01 and 0.001 levels of confidence. Square = AIS; triangle = IS.
Figure Legend Snippet: Changes in the size and fine structure of chloroplasts. Size and fine structures of chloroplasts was evaluated by TEM on longitudinal ultrathin sections within the mesophyll of leaves from Arabidopsis thaliana Col-0 leaves inoculated with Botrytis cinerea and compared to CL. Measurements were performed 0 h ( = CL, represented by the dotted line), 12, 24, 48 and 96 hpi at the IS and the AIS. Shown are means with standard errors. n > 100 chloroplasts per treatment. Significant differences were calculated using the Mann-Whitney U-test; *, **, and ***, respectively, indicate significance at the 0.05, 0.01 and 0.001 levels of confidence. Square = AIS; triangle = IS.

Techniques Used: Transmission Electron Microscopy, MANN-WHITNEY

7) Product Images from "Genus-optimized strategy for the identification of chlamydial type III secretion substrates"

Article Title: Genus-optimized strategy for the identification of chlamydial type III secretion substrates

Journal: Pathogens and disease

doi: 10.1111/2049-632X.12070

CT082 localizes to the surface of chlamydiae during infection C. trachomatis -infected Hela cells were fixed and embedded in unicryl resin at 24 hr (A to F) or 42 hr (G to L) post infection. Ultrathin sections were cut, collected on Ni grids and labeled with guinea pig anti-CT082 antibody and 10 nm gold-conjugated goat anti guinea pig IgG. No gold particles were observed when sections were labeled with gold-conjugated antibody only (not shown). Representative RBs (A to F) and EBs (G to L) from different inclusions are shown here. In all panels, the scale bar is 100nm.
Figure Legend Snippet: CT082 localizes to the surface of chlamydiae during infection C. trachomatis -infected Hela cells were fixed and embedded in unicryl resin at 24 hr (A to F) or 42 hr (G to L) post infection. Ultrathin sections were cut, collected on Ni grids and labeled with guinea pig anti-CT082 antibody and 10 nm gold-conjugated goat anti guinea pig IgG. No gold particles were observed when sections were labeled with gold-conjugated antibody only (not shown). Representative RBs (A to F) and EBs (G to L) from different inclusions are shown here. In all panels, the scale bar is 100nm.

Techniques Used: Infection, Labeling

8) Product Images from "Aim24 and MICOS modulate respiratory function, tafazzin-related cardiolipin modification and mitochondrial architecture"

Article Title: Aim24 and MICOS modulate respiratory function, tafazzin-related cardiolipin modification and mitochondrial architecture

Journal: eLife

doi: 10.7554/eLife.01684

Deletion of the AIM24 gene leads to altered mitochondrial morphology and architecture. ( A and B ) Electron microscopy of wild type and AIM24 deletion mutant. Cells were grown in either YPD or YPG and fixed with glutaraldehyde, contrasted with osmium tetroxide and uranylacetate. Ultrathin sections were obtained and contrasted with uranylacetate and lead citrate. ( A ) Wild type. a–c, cells grown on YPD; d–f cells grown on YPG; a and d, cells (scale bar, 1 µm); b, c, e, f, mitochondrial profiles (scale bar, 0.2 µm). ( B ) Δ aim24 . a–i, cells grown on YPG; j–o cells grown on YPD; a, b, j and k, cells (scale bar, 1 µm). c–i and l–o, mitochondrial profiles (scale bars, 0.2 µm). ( C ) Fluorescence microscopy of wild type and Δ aim24 cells expressing mitoGFP. Cells were grown at 30°C on YPD or YPG medium to stationary phase (scale bar, 5 µm). DOI: http://dx.doi.org/10.7554/eLife.01684.010
Figure Legend Snippet: Deletion of the AIM24 gene leads to altered mitochondrial morphology and architecture. ( A and B ) Electron microscopy of wild type and AIM24 deletion mutant. Cells were grown in either YPD or YPG and fixed with glutaraldehyde, contrasted with osmium tetroxide and uranylacetate. Ultrathin sections were obtained and contrasted with uranylacetate and lead citrate. ( A ) Wild type. a–c, cells grown on YPD; d–f cells grown on YPG; a and d, cells (scale bar, 1 µm); b, c, e, f, mitochondrial profiles (scale bar, 0.2 µm). ( B ) Δ aim24 . a–i, cells grown on YPG; j–o cells grown on YPD; a, b, j and k, cells (scale bar, 1 µm). c–i and l–o, mitochondrial profiles (scale bars, 0.2 µm). ( C ) Fluorescence microscopy of wild type and Δ aim24 cells expressing mitoGFP. Cells were grown at 30°C on YPD or YPG medium to stationary phase (scale bar, 5 µm). DOI: http://dx.doi.org/10.7554/eLife.01684.010

Techniques Used: Electron Microscopy, Mutagenesis, Fluorescence, Microscopy, Expressing

9) Product Images from "Agitation Modules: Flexible Means to Accelerate Automated Freeze Substitution"

Article Title: Agitation Modules: Flexible Means to Accelerate Automated Freeze Substitution

Journal: Journal of Histochemistry and Cytochemistry

doi: 10.1369/0022155418786698

Immunogold labeling of Lowicryl HM20 sections of HP-frozen, low-temperature embedded immature anthers of Arabidopsis thaliana generated by accelerated FS under agitation. (A) Overview image showing a middle layer cell from an anther surrounded by anti-xyloglucan labeled cell wall (cw). (B) Aggregation of intracellular vesicle-like xyloglucan-positive structures. (C) Overview image showing anti-α-tubulin labeling close to the plasma membrane. (D) Details of immunolabeled cortical microtubules (arrows) aligned almost in parallel to the section plane. Typical for postembedding labeling, the gold-conjugated antibodies are concentrated at regions where the MTs meet the surface of the ultrathin section. Scale bars = 1 µm in A and C; 300 nm in B and D. Abbreviations: n, nucleus; v, vacuole; chl, chloroplast; HP, high-pressure; FS, freeze substitution; MT, microtubule.
Figure Legend Snippet: Immunogold labeling of Lowicryl HM20 sections of HP-frozen, low-temperature embedded immature anthers of Arabidopsis thaliana generated by accelerated FS under agitation. (A) Overview image showing a middle layer cell from an anther surrounded by anti-xyloglucan labeled cell wall (cw). (B) Aggregation of intracellular vesicle-like xyloglucan-positive structures. (C) Overview image showing anti-α-tubulin labeling close to the plasma membrane. (D) Details of immunolabeled cortical microtubules (arrows) aligned almost in parallel to the section plane. Typical for postembedding labeling, the gold-conjugated antibodies are concentrated at regions where the MTs meet the surface of the ultrathin section. Scale bars = 1 µm in A and C; 300 nm in B and D. Abbreviations: n, nucleus; v, vacuole; chl, chloroplast; HP, high-pressure; FS, freeze substitution; MT, microtubule.

Techniques Used: Labeling, Generated, Immunolabeling

10) Product Images from "Ciliary neurotrophic factor promotes motor reinnervation of the musculocutaneous nerve in an experimental model of end-to-side neurorrhaphy"

Article Title: Ciliary neurotrophic factor promotes motor reinnervation of the musculocutaneous nerve in an experimental model of end-to-side neurorrhaphy

Journal: BMC Neuroscience

doi: 10.1186/1471-2202-12-58

Representative ultrathin sections through MCN . Electron micrographs showing representative myelinated axons in cross sections through intact MCN (A), and MCN stumps 2 months after their reconnection with the UN and intrathecal application of vehiculum (B), Cerebrolysin (C) and CNTF (D). Scale bars = 2 μm.
Figure Legend Snippet: Representative ultrathin sections through MCN . Electron micrographs showing representative myelinated axons in cross sections through intact MCN (A), and MCN stumps 2 months after their reconnection with the UN and intrathecal application of vehiculum (B), Cerebrolysin (C) and CNTF (D). Scale bars = 2 μm.

Techniques Used:

11) Product Images from "Selective Degradation Permits a Feedback Loop Controlling Annexin A6 and Cholesterol Levels in Endolysosomes of NPC1 Mutant Cells"

Article Title: Selective Degradation Permits a Feedback Loop Controlling Annexin A6 and Cholesterol Levels in Endolysosomes of NPC1 Mutant Cells

Journal: Cells

doi: 10.3390/cells9051152

AnxA6 is enriched in inner membranes of the late endosome/lysosome (LE/Lys) compartment in NPC1 mutant cells. Lowicryl HM20 ultrathin sections of CHO-WT and CHO M12 cells were labeled with anti-AnxA6. ( A ) A region of the cytoplasm of CHO-WT and CHO M12 cells with several LE/Lys structures is shown. Note that in CHO M12 cells AnxA6 gold labeling is much more prominent compared to CHO-WT cells. Encircled gold particles (dashed yellow) indicate cytoplasmatic AnxA6 labeling. ( B ) Quantification of gold particle density (gold particle/μm 2 ) and LE/Lys vesicle area (μm 2 ). The data is shown as dot plot per cell. The mean ± SEM is also given (*** p
Figure Legend Snippet: AnxA6 is enriched in inner membranes of the late endosome/lysosome (LE/Lys) compartment in NPC1 mutant cells. Lowicryl HM20 ultrathin sections of CHO-WT and CHO M12 cells were labeled with anti-AnxA6. ( A ) A region of the cytoplasm of CHO-WT and CHO M12 cells with several LE/Lys structures is shown. Note that in CHO M12 cells AnxA6 gold labeling is much more prominent compared to CHO-WT cells. Encircled gold particles (dashed yellow) indicate cytoplasmatic AnxA6 labeling. ( B ) Quantification of gold particle density (gold particle/μm 2 ) and LE/Lys vesicle area (μm 2 ). The data is shown as dot plot per cell. The mean ± SEM is also given (*** p

Techniques Used: Mutagenesis, Labeling

12) Product Images from "Agitation Modules: Flexible Means to Accelerate Automated Freeze Substitution"

Article Title: Agitation Modules: Flexible Means to Accelerate Automated Freeze Substitution

Journal: Journal of Histochemistry and Cytochemistry

doi: 10.1369/0022155418786698

Immunogold labeling of Lowicryl HM20 sections of HP-frozen, low-temperature embedded immature anthers of Arabidopsis thaliana generated by accelerated FS under agitation. (A) Overview image showing a middle layer cell from an anther surrounded by anti-xyloglucan labeled cell wall (cw). (B) Aggregation of intracellular vesicle-like xyloglucan-positive structures. (C) Overview image showing anti-α-tubulin labeling close to the plasma membrane. (D) Details of immunolabeled cortical microtubules (arrows) aligned almost in parallel to the section plane. Typical for postembedding labeling, the gold-conjugated antibodies are concentrated at regions where the MTs meet the surface of the ultrathin section. Scale bars = 1 µm in A and C; 300 nm in B and D. Abbreviations: n, nucleus; v, vacuole; chl, chloroplast; HP, high-pressure; FS, freeze substitution; MT, microtubule.
Figure Legend Snippet: Immunogold labeling of Lowicryl HM20 sections of HP-frozen, low-temperature embedded immature anthers of Arabidopsis thaliana generated by accelerated FS under agitation. (A) Overview image showing a middle layer cell from an anther surrounded by anti-xyloglucan labeled cell wall (cw). (B) Aggregation of intracellular vesicle-like xyloglucan-positive structures. (C) Overview image showing anti-α-tubulin labeling close to the plasma membrane. (D) Details of immunolabeled cortical microtubules (arrows) aligned almost in parallel to the section plane. Typical for postembedding labeling, the gold-conjugated antibodies are concentrated at regions where the MTs meet the surface of the ultrathin section. Scale bars = 1 µm in A and C; 300 nm in B and D. Abbreviations: n, nucleus; v, vacuole; chl, chloroplast; HP, high-pressure; FS, freeze substitution; MT, microtubule.

Techniques Used: Labeling, Generated, Immunolabeling

13) Product Images from "Mapping synaptic input fields of neurons with super-resolution imaging"

Article Title: Mapping synaptic input fields of neurons with super-resolution imaging

Journal: Cell

doi: 10.1016/j.cell.2015.08.033

A super-resolution imaging and analysis platform (A) Tissues were dissected, fixed for immunohistochemical labeling, postfixed, dehydrated, and embedded in epoxy resin. Ultrathin sections were cut, arrayed on glass coverslips, and etched to expose fluorophores for STORM imaging. Individual serial sections were imaged and aligned to generate 3D reconstructions. (B) STORM maximum intensity projection of a volume (2.3 × 10 5 μm 3 ) of the mouse IPL containing an On-Off DSGC (blue) amidst presynaptic (magenta) and gephyrin (green) clusters imaged using the platform. (C) .
Figure Legend Snippet: A super-resolution imaging and analysis platform (A) Tissues were dissected, fixed for immunohistochemical labeling, postfixed, dehydrated, and embedded in epoxy resin. Ultrathin sections were cut, arrayed on glass coverslips, and etched to expose fluorophores for STORM imaging. Individual serial sections were imaged and aligned to generate 3D reconstructions. (B) STORM maximum intensity projection of a volume (2.3 × 10 5 μm 3 ) of the mouse IPL containing an On-Off DSGC (blue) amidst presynaptic (magenta) and gephyrin (green) clusters imaged using the platform. (C) .

Techniques Used: Imaging, Immunohistochemistry, Labeling

14) Product Images from "Hydrogels of Polycationic Acetohydrazone-Modified Phosphorus Dendrimers for Biomedical Applications: Gelation Studies and Nucleic Acid Loading"

Article Title: Hydrogels of Polycationic Acetohydrazone-Modified Phosphorus Dendrimers for Biomedical Applications: Gelation Studies and Nucleic Acid Loading

Journal: Pharmaceutics

doi: 10.3390/pharmaceutics10030120

TEM images of hydrogel networks formed from dendrimer solutions in water: TG2 @10% Glc ( a , b ); TG3 @10% Glc ( c ); PG2 @10% Glc ( d ); PG3 @10% Glc ( e ); PG1 @10% Gly ( f ); PG2 @10% Gly ( g ); PG3 @10% Gly ( h ); PG3 @10% PEG ( i ). Ultrathin sections (~70 nm thick). Scale bars represent 100 nm.
Figure Legend Snippet: TEM images of hydrogel networks formed from dendrimer solutions in water: TG2 @10% Glc ( a , b ); TG3 @10% Glc ( c ); PG2 @10% Glc ( d ); PG3 @10% Glc ( e ); PG1 @10% Gly ( f ); PG2 @10% Gly ( g ); PG3 @10% Gly ( h ); PG3 @10% PEG ( i ). Ultrathin sections (~70 nm thick). Scale bars represent 100 nm.

Techniques Used: Transmission Electron Microscopy, Gas Chromatography

15) Product Images from "A complex virome that includes two distinct emaraviruses is associated to virus-like symptoms in Camellia japonica"

Article Title: A complex virome that includes two distinct emaraviruses is associated to virus-like symptoms in Camellia japonica

Journal: bioRxiv

doi: 10.1101/822254

Coiled virus-like particles observed in negative staining of crude extract (a, b, c, d) and spherical double-enveloped bodies in ultrathin sections (e, f, g) of symptomatic camellia leaves. Scale bar: 100 nm
Figure Legend Snippet: Coiled virus-like particles observed in negative staining of crude extract (a, b, c, d) and spherical double-enveloped bodies in ultrathin sections (e, f, g) of symptomatic camellia leaves. Scale bar: 100 nm

Techniques Used: Negative Staining

16) Product Images from "Agitation Modules: Flexible Means to Accelerate Automated Freeze Substitution"

Article Title: Agitation Modules: Flexible Means to Accelerate Automated Freeze Substitution

Journal: Journal of Histochemistry and Cytochemistry

doi: 10.1369/0022155418786698

Immunogold labeling of Lowicryl HM20 sections of HP-frozen, low-temperature embedded immature anthers of Arabidopsis thaliana generated by accelerated FS under agitation. (A) Overview image showing a middle layer cell from an anther surrounded by anti-xyloglucan labeled cell wall (cw). (B) Aggregation of intracellular vesicle-like xyloglucan-positive structures. (C) Overview image showing anti-α-tubulin labeling close to the plasma membrane. (D) Details of immunolabeled cortical microtubules (arrows) aligned almost in parallel to the section plane. Typical for postembedding labeling, the gold-conjugated antibodies are concentrated at regions where the MTs meet the surface of the ultrathin section. Scale bars = 1 µm in A and C; 300 nm in B and D. Abbreviations: n, nucleus; v, vacuole; chl, chloroplast; HP, high-pressure; FS, freeze substitution; MT, microtubule.
Figure Legend Snippet: Immunogold labeling of Lowicryl HM20 sections of HP-frozen, low-temperature embedded immature anthers of Arabidopsis thaliana generated by accelerated FS under agitation. (A) Overview image showing a middle layer cell from an anther surrounded by anti-xyloglucan labeled cell wall (cw). (B) Aggregation of intracellular vesicle-like xyloglucan-positive structures. (C) Overview image showing anti-α-tubulin labeling close to the plasma membrane. (D) Details of immunolabeled cortical microtubules (arrows) aligned almost in parallel to the section plane. Typical for postembedding labeling, the gold-conjugated antibodies are concentrated at regions where the MTs meet the surface of the ultrathin section. Scale bars = 1 µm in A and C; 300 nm in B and D. Abbreviations: n, nucleus; v, vacuole; chl, chloroplast; HP, high-pressure; FS, freeze substitution; MT, microtubule.

Techniques Used: Labeling, Generated, Immunolabeling

17) Product Images from "High Resolution Imaging of Temporal and Spatial Changes of Subcellular Ascorbate, Glutathione and H2O2 Distribution during Botrytis cinerea Infection in Arabidopsis"

Article Title: High Resolution Imaging of Temporal and Spatial Changes of Subcellular Ascorbate, Glutathione and H2O2 Distribution during Botrytis cinerea Infection in Arabidopsis

Journal: PLoS ONE

doi: 10.1371/journal.pone.0065811

Changes in the size and fine structure of chloroplasts. Size and fine structures of chloroplasts was evaluated by TEM on longitudinal ultrathin sections within the mesophyll of leaves from Arabidopsis thaliana Col-0 leaves inoculated with Botrytis cinerea and compared to CL. Measurements were performed 0 h ( = CL, represented by the dotted line), 12, 24, 48 and 96 hpi at the IS and the AIS. Shown are means with standard errors. n > 100 chloroplasts per treatment. Significant differences were calculated using the Mann-Whitney U-test; *, **, and ***, respectively, indicate significance at the 0.05, 0.01 and 0.001 levels of confidence. Square = AIS; triangle = IS.
Figure Legend Snippet: Changes in the size and fine structure of chloroplasts. Size and fine structures of chloroplasts was evaluated by TEM on longitudinal ultrathin sections within the mesophyll of leaves from Arabidopsis thaliana Col-0 leaves inoculated with Botrytis cinerea and compared to CL. Measurements were performed 0 h ( = CL, represented by the dotted line), 12, 24, 48 and 96 hpi at the IS and the AIS. Shown are means with standard errors. n > 100 chloroplasts per treatment. Significant differences were calculated using the Mann-Whitney U-test; *, **, and ***, respectively, indicate significance at the 0.05, 0.01 and 0.001 levels of confidence. Square = AIS; triangle = IS.

Techniques Used: Transmission Electron Microscopy, MANN-WHITNEY

18) Product Images from "Extracellular Vesicles Mediate Receptor-Independent Transmission of Novel Tick-Borne Bunyavirus"

Article Title: Extracellular Vesicles Mediate Receptor-Independent Transmission of Novel Tick-Borne Bunyavirus

Journal: Journal of Virology

doi: 10.1128/JVI.02490-15

SFTS virus NSs induces the formation of endosome-like structures. Ultrastructure analyses of SFTS virus NSs-expressing cells (A) and SFTS virus-infected cells (B) show cytoplasmic structures reminiscent of early endosomes (arrows) in ultrathin sections. Immunogold staining of distinct ultrathin sections shows cytoplasmic structures positive for SFTS virus NSs and Rab5 (C) or SFTS virus NSs and LC3B (D). A solid red arrows indicate SFTS virus NSs in panels C and D, while a red triangle (in panel C) and a green triangle (in panel D) indicate the detection of Rab5 and LC3B, respectively. Ultrathin sections of mock-infected cells were also labeled, as indicated above (not shown), to ensure the specificity of antibody. Representatives images are shown.
Figure Legend Snippet: SFTS virus NSs induces the formation of endosome-like structures. Ultrastructure analyses of SFTS virus NSs-expressing cells (A) and SFTS virus-infected cells (B) show cytoplasmic structures reminiscent of early endosomes (arrows) in ultrathin sections. Immunogold staining of distinct ultrathin sections shows cytoplasmic structures positive for SFTS virus NSs and Rab5 (C) or SFTS virus NSs and LC3B (D). A solid red arrows indicate SFTS virus NSs in panels C and D, while a red triangle (in panel C) and a green triangle (in panel D) indicate the detection of Rab5 and LC3B, respectively. Ultrathin sections of mock-infected cells were also labeled, as indicated above (not shown), to ensure the specificity of antibody. Representatives images are shown.

Techniques Used: Expressing, Infection, Staining, Labeling

19) Product Images from "Clustered granules present in the hippocampus of aged mice result from a degenerative process affecting astrocytes and their surrounding neuropil"

Article Title: Clustered granules present in the hippocampus of aged mice result from a degenerative process affecting astrocytes and their surrounding neuropil

Journal: Age

doi: 10.1007/s11357-014-9690-8

Immunoelectron microscopy images of hippocampal granules of 14-month-old SAMP8 mice stained with IgM antibodies directed against the neo-epitope. a A granule of an ultrathin section embedded in Lowicryl is shown. The staining of the antibody is abundantly
Figure Legend Snippet: Immunoelectron microscopy images of hippocampal granules of 14-month-old SAMP8 mice stained with IgM antibodies directed against the neo-epitope. a A granule of an ultrathin section embedded in Lowicryl is shown. The staining of the antibody is abundantly

Techniques Used: Immuno-Electron Microscopy, Mouse Assay, Staining

20) Product Images from "Coliform Bacteria for Bioremediation of Waste Hydrocarbons"

Article Title: Coliform Bacteria for Bioremediation of Waste Hydrocarbons

Journal: BioMed Research International

doi: 10.1155/2017/1838072

Transmission electron micrographs of ultrathin sections of E . coli cells from domestic sewage grown in n -hexadecane-free medium (a) and in n -hexadecane-containing medium (b) and of E . coli cells from wheat straw grown in n -hexadecane-free medium (c) and in n -hexadecane-containing medium (d). Magnification: (a) and (b) ×60000; (c) and (d) ×40000.
Figure Legend Snippet: Transmission electron micrographs of ultrathin sections of E . coli cells from domestic sewage grown in n -hexadecane-free medium (a) and in n -hexadecane-containing medium (b) and of E . coli cells from wheat straw grown in n -hexadecane-free medium (c) and in n -hexadecane-containing medium (d). Magnification: (a) and (b) ×60000; (c) and (d) ×40000.

Techniques Used: Transmission Assay

21) Product Images from "Morphology of Mitochondria in Syncytial Annelid Female Germ-Line Cyst Visualized by Serial Block-Face SEM"

Article Title: Morphology of Mitochondria in Syncytial Annelid Female Germ-Line Cyst Visualized by Serial Block-Face SEM

Journal: International Journal of Cell Biology

doi: 10.1155/2020/7483467

Organization of the mitochondria in syncytial germ-line cysts. (a) and (b) The accumulations of mitochondria ( stars ) are located mainly in the one pole of the nurse cells (NC) opposite the ring canal ( bilateral dotted arrow in (b)), while in the cytophore ( Cy ) the mitochondria were evenly scattered in the cytoplasm. Nuclei of the nurse cells and oocyte ( O ) are marked as Nu ; tubules of the endoplasmic reticulum as ER . (c) and (d) the mitochondria (stained black) are visible passing through the ring canals ( arrows ) connecting the nurse cells ( NC ) to the cytophore ( Cy ). Nu – nurse cell nucleus. (a, c, d) Light microscopy, Epon semi-thin sections stained with methylene blue. (a) Bar = 73 µ m; (c, d) bar = 30 µ m. (b) Transmission electron microscopy, Epon ultrathin section; bar = 2 µ m.
Figure Legend Snippet: Organization of the mitochondria in syncytial germ-line cysts. (a) and (b) The accumulations of mitochondria ( stars ) are located mainly in the one pole of the nurse cells (NC) opposite the ring canal ( bilateral dotted arrow in (b)), while in the cytophore ( Cy ) the mitochondria were evenly scattered in the cytoplasm. Nuclei of the nurse cells and oocyte ( O ) are marked as Nu ; tubules of the endoplasmic reticulum as ER . (c) and (d) the mitochondria (stained black) are visible passing through the ring canals ( arrows ) connecting the nurse cells ( NC ) to the cytophore ( Cy ). Nu – nurse cell nucleus. (a, c, d) Light microscopy, Epon semi-thin sections stained with methylene blue. (a) Bar = 73 µ m; (c, d) bar = 30 µ m. (b) Transmission electron microscopy, Epon ultrathin section; bar = 2 µ m.

Techniques Used: Staining, Light Microscopy, Transmission Assay, Electron Microscopy

(a–c) Accumulations of mitochondria ( Mi ) together with the tubules of the endoplasmic reticulum ( ER ) that enclosed the Golgi complexes ( GC ) are visible in the nurse cell pole opposite the ring canal, near the cell nucleus ( Nu ). Note the two distinguishable kinds of mitochondria with electron-dense and electron-lucent matrix, both of them possessing numerous cristae. Granulo-fibrillar nuage material ( arrows ) was observed close to mitochondrial aggregation within both nurse cells and cytophore (inset). (d) Thioflavin T staining showed that the nuage material was rich in amyloid-like proteins ( arrows ). NC – nurse cells; Nu – nurse cell nucleus. (e–i) Autophagosomes in nurse cells containing the remnants of cell organelles including the mitochondria ( arrows ). Tubules of endoplasmic reticulum – ER ; Golgi complexes – GC . (a–c), (e–i) Transmission electron microscopy, Epon ultrathin sections; (a) bar = 1.4 µ m; (b), (c), (e) bar = 0.9 µ m; (f) bar = 0.5 µ m; (g, h) bar = 0.4 µ m; (i) bar = 0.3 µ m. (d) Steedman wax sections stained with thioflavin-T; bar = 45 µ m.
Figure Legend Snippet: (a–c) Accumulations of mitochondria ( Mi ) together with the tubules of the endoplasmic reticulum ( ER ) that enclosed the Golgi complexes ( GC ) are visible in the nurse cell pole opposite the ring canal, near the cell nucleus ( Nu ). Note the two distinguishable kinds of mitochondria with electron-dense and electron-lucent matrix, both of them possessing numerous cristae. Granulo-fibrillar nuage material ( arrows ) was observed close to mitochondrial aggregation within both nurse cells and cytophore (inset). (d) Thioflavin T staining showed that the nuage material was rich in amyloid-like proteins ( arrows ). NC – nurse cells; Nu – nurse cell nucleus. (e–i) Autophagosomes in nurse cells containing the remnants of cell organelles including the mitochondria ( arrows ). Tubules of endoplasmic reticulum – ER ; Golgi complexes – GC . (a–c), (e–i) Transmission electron microscopy, Epon ultrathin sections; (a) bar = 1.4 µ m; (b), (c), (e) bar = 0.9 µ m; (f) bar = 0.5 µ m; (g, h) bar = 0.4 µ m; (i) bar = 0.3 µ m. (d) Steedman wax sections stained with thioflavin-T; bar = 45 µ m.

Techniques Used: Staining, Transmission Assay, Electron Microscopy

22) Product Images from "The integrity of cochlear hair cells is established and maintained through the localization of Dia1 at apical junctional complexes and stereocilia"

Article Title: The integrity of cochlear hair cells is established and maintained through the localization of Dia1 at apical junctional complexes and stereocilia

Journal: Cell Death & Disease

doi: 10.1038/s41419-020-02743-z

Ultrastructural analysis of the organ of Corti in 5-month-old DIA1 -TG mice by transmission electron microscopy. Five-month-old WT ( a–g ) and DIA1 TG/TG (TG) ( h–n ) mice were fixed for scanning electron microscopy (SEM) and transmission electron microscopy (TEM). a, h SEM images of OHCs and supporting cells (SCs). OHC loss (dots) and abnormal, such as short and sparse (arrows), stereocilia in TG mice. b , i Peripheral nerves at the osseous spiral lamina at the basal turn of the cochlea in WT ( b ) and TG mice ( i ). c–g, j–n High magnification views of ultrathin section of OHCs at the distal portion of the middle turn (close to the basal turn) of the cochlea by TEM. Magnification views of apical junctional complexes (AJCs) and cuticular plates of OHCs in WT ( c ) and TG ( j ) mice were shown in d – g and k – n , respectively. Note the hazy (arrows in k – m ), ruffled peri-junctional actin belts (arrows in n ) and cuticular plates (double arrow in n ) of OHCs in TG mice, compared to smooth actin belts (dotted lines) in WT mice ( d – g ). Asterisk and double asterisks indicate the same OHCs, respectively. Scale bar: 5 μm ( a , h ), 2 μm ( b , c , i , j ), and 500 nm ( d – g , k – n ).
Figure Legend Snippet: Ultrastructural analysis of the organ of Corti in 5-month-old DIA1 -TG mice by transmission electron microscopy. Five-month-old WT ( a–g ) and DIA1 TG/TG (TG) ( h–n ) mice were fixed for scanning electron microscopy (SEM) and transmission electron microscopy (TEM). a, h SEM images of OHCs and supporting cells (SCs). OHC loss (dots) and abnormal, such as short and sparse (arrows), stereocilia in TG mice. b , i Peripheral nerves at the osseous spiral lamina at the basal turn of the cochlea in WT ( b ) and TG mice ( i ). c–g, j–n High magnification views of ultrathin section of OHCs at the distal portion of the middle turn (close to the basal turn) of the cochlea by TEM. Magnification views of apical junctional complexes (AJCs) and cuticular plates of OHCs in WT ( c ) and TG ( j ) mice were shown in d – g and k – n , respectively. Note the hazy (arrows in k – m ), ruffled peri-junctional actin belts (arrows in n ) and cuticular plates (double arrow in n ) of OHCs in TG mice, compared to smooth actin belts (dotted lines) in WT mice ( d – g ). Asterisk and double asterisks indicate the same OHCs, respectively. Scale bar: 5 μm ( a , h ), 2 μm ( b , c , i , j ), and 500 nm ( d – g , k – n ).

Techniques Used: Mouse Assay, Transmission Assay, Electron Microscopy, Transmission Electron Microscopy

23) Product Images from "Midbody and primary cilium of neural progenitors release extracellular membrane particles enriched in the stem cell marker prominin-1"

Article Title: Midbody and primary cilium of neural progenitors release extracellular membrane particles enriched in the stem cell marker prominin-1

Journal: The Journal of Cell Biology

doi: 10.1083/jcb.200608137

Prom1 particles contain α-tubulin. (A) Neural tube fluid of E11.5 mouse embryos was subjected to differential centrifugation ( Marzesco et al., 2005 ) followed by immunoblot analysis of the fractions for α-tubulin. P1, 300- g pellet; P2, 1,200- g pellet; P3, 10,000- g pellet; P4, 100,000- g pellet; S4, 100,000- g supernatant. (B) Transverse cryosection of E10.5 mouse telencephalon double immunostained for prom1 (red) and α-tubulin (green) and analyzed by confocal microscopy; four examples of prom1 particles in the lumen of the telencephalic ventricle are shown (z-stack projection). (C and D) Transverse ultrathin cryosections of the apical region of mouse E10.5 telencephalic NE cells immunogold labeled for prom1 (5 nm) and α-tubulin (12 nm) showing two doubly immunoreactive particles at the lumenal surface. (E) Negative staining of a particle from the P2 pellet after immunogold labeling for prom1 (5 nm) and acetylated tubulin (12 nm). Bars: (B) 1 μm; (C–E) 100 nm.
Figure Legend Snippet: Prom1 particles contain α-tubulin. (A) Neural tube fluid of E11.5 mouse embryos was subjected to differential centrifugation ( Marzesco et al., 2005 ) followed by immunoblot analysis of the fractions for α-tubulin. P1, 300- g pellet; P2, 1,200- g pellet; P3, 10,000- g pellet; P4, 100,000- g pellet; S4, 100,000- g supernatant. (B) Transverse cryosection of E10.5 mouse telencephalon double immunostained for prom1 (red) and α-tubulin (green) and analyzed by confocal microscopy; four examples of prom1 particles in the lumen of the telencephalic ventricle are shown (z-stack projection). (C and D) Transverse ultrathin cryosections of the apical region of mouse E10.5 telencephalic NE cells immunogold labeled for prom1 (5 nm) and α-tubulin (12 nm) showing two doubly immunoreactive particles at the lumenal surface. (E) Negative staining of a particle from the P2 pellet after immunogold labeling for prom1 (5 nm) and acetylated tubulin (12 nm). Bars: (B) 1 μm; (C–E) 100 nm.

Techniques Used: Centrifugation, Confocal Microscopy, Labeling, Negative Staining

Anillin is present not only in prom1-bearing midbodies but also in prom1-bearing lumenal particles. (A and B) Transverse ultrathin cryosections of the apical surface of E10.5 mouse telencephalic neuroepithelium double immunogold labeled for prom1 (5 nm) and anillin (10 nm; white arrowheads). Doubly immunoreactive midbody (A) and particle at the apical surface of the neuroepithelium (B). Black arrowheads indicate prom1-labeled regions at the lumenal plasma membrane of the midbody. (C) Double immunogold labeling for prom1 (5 nm) and anillin (10 nm; white arrowheads) of a particle from the P2 pellet of E11.5 neural tube fluid. (D–F) Transverse cryosections of E11.5 (D and E) and E10.5 (F) mouse telencephalon double immunostained for prom1 (red) and anillin (green) and analyzed by confocal microscopy. (D and E) Z-stack projection providing an en face view onto the apical surface of the neuroepithelium; for orientation, the DAPI staining of nuclei (blue) is shown for one of the optical sections in D. (E) Selected regions of D (indicated by arrowheads) at higher magnification. (F) Four examples of prom1-bearing particles in the lumen of the telencephalic ventricle (z-stack projection). Bars: (A–C) 100 nm; (D) 10 μm; (F) 1 μm.
Figure Legend Snippet: Anillin is present not only in prom1-bearing midbodies but also in prom1-bearing lumenal particles. (A and B) Transverse ultrathin cryosections of the apical surface of E10.5 mouse telencephalic neuroepithelium double immunogold labeled for prom1 (5 nm) and anillin (10 nm; white arrowheads). Doubly immunoreactive midbody (A) and particle at the apical surface of the neuroepithelium (B). Black arrowheads indicate prom1-labeled regions at the lumenal plasma membrane of the midbody. (C) Double immunogold labeling for prom1 (5 nm) and anillin (10 nm; white arrowheads) of a particle from the P2 pellet of E11.5 neural tube fluid. (D–F) Transverse cryosections of E11.5 (D and E) and E10.5 (F) mouse telencephalon double immunostained for prom1 (red) and anillin (green) and analyzed by confocal microscopy. (D and E) Z-stack projection providing an en face view onto the apical surface of the neuroepithelium; for orientation, the DAPI staining of nuclei (blue) is shown for one of the optical sections in D. (E) Selected regions of D (indicated by arrowheads) at higher magnification. (F) Four examples of prom1-bearing particles in the lumen of the telencephalic ventricle (z-stack projection). Bars: (A–C) 100 nm; (D) 10 μm; (F) 1 μm.

Techniques Used: Labeling, Confocal Microscopy, Staining

Ultrastructural resemblance of lumenal particles to aged midbodies of NE cells. EM analysis of serial ultrathin (70 nm) plastic sections of the apical surface of E10.5 mouse telencephalic neuroepithelium. (A) Midbody connecting NE daughter cells in telophase. (B–D) Series showing every other section of an aged midbody. (E) Midbody of an NE cell that has relocated the centriole (arrow) apically. (F–H) Series showing every third section of an electron-dense particle detached from the apical surface of the neuroepithelium. Arrowheads indicate plasma membrane buds and protrusions, and asterisks indicate adherens junctions. The complete sequence of serial sections from which A, E, and F–H were selected is shown in Fig. S4 (A–C), respectively (available at http://www.jcb.org/cgi/content/full/jcb.200608137/DC1 ). Bars, 500 nm.
Figure Legend Snippet: Ultrastructural resemblance of lumenal particles to aged midbodies of NE cells. EM analysis of serial ultrathin (70 nm) plastic sections of the apical surface of E10.5 mouse telencephalic neuroepithelium. (A) Midbody connecting NE daughter cells in telophase. (B–D) Series showing every other section of an aged midbody. (E) Midbody of an NE cell that has relocated the centriole (arrow) apically. (F–H) Series showing every third section of an electron-dense particle detached from the apical surface of the neuroepithelium. Arrowheads indicate plasma membrane buds and protrusions, and asterisks indicate adherens junctions. The complete sequence of serial sections from which A, E, and F–H were selected is shown in Fig. S4 (A–C), respectively (available at http://www.jcb.org/cgi/content/full/jcb.200608137/DC1 ). Bars, 500 nm.

Techniques Used: Sequencing

Prom1 is concentrated at the midbody of NE cells. Mouse embryonic forebrain was subjected to preembedding immunogold labeling for prom1 (10 nm gold) followed by EM analysis of ultrathin serial plastic sections. Apical midbodies of NE cells at E8.5 (A–E), E10.5 (H–J), and E12.5 (F and G) are shown. (B and C) The central portion of the midbody with long, thin stalks depicted in B is shown at higher magnification of a consecutive section in C. (D and E) Consecutive sections showing the central portion of another midbody with long, thin stalks. (G–I) Midbodies with short stalks. (J) Particle at the apical surface of a neuroephithelial cell. Black arrowheads indicate prom1-labeled plasma membrane buds and protrusions, gray arrowheads indicate detached particles, solid arrows indicate midbody plasma membrane facing the lumenal side, and open arrows indicate midbody plasma membrane derived from cleavage furrow. Bars: (A and C–J) 100 nm; (B) 1 μm.
Figure Legend Snippet: Prom1 is concentrated at the midbody of NE cells. Mouse embryonic forebrain was subjected to preembedding immunogold labeling for prom1 (10 nm gold) followed by EM analysis of ultrathin serial plastic sections. Apical midbodies of NE cells at E8.5 (A–E), E10.5 (H–J), and E12.5 (F and G) are shown. (B and C) The central portion of the midbody with long, thin stalks depicted in B is shown at higher magnification of a consecutive section in C. (D and E) Consecutive sections showing the central portion of another midbody with long, thin stalks. (G–I) Midbodies with short stalks. (J) Particle at the apical surface of a neuroephithelial cell. Black arrowheads indicate prom1-labeled plasma membrane buds and protrusions, gray arrowheads indicate detached particles, solid arrows indicate midbody plasma membrane facing the lumenal side, and open arrows indicate midbody plasma membrane derived from cleavage furrow. Bars: (A and C–J) 100 nm; (B) 1 μm.

Techniques Used: Labeling, Derivative Assay

Prom1 is concentrated on cilia of NE cells. Mouse forebrain was subjected to preembedding immunogold labeling for prom1 (10 nm gold) followed by EM analysis of ultrathin plastic sections. (A) A cilium at E8.5 showing weak labeling (arrows). (B–F) Cilia at E10.5 (B and C), E12.5 (D and F), or E13.5 (E) showing different degrees of surface labeling; note the cluster of prom1 at the tip of the cilium in D and E (arrowheads). (G and H) Short cilia at E12.5 with strongly prom1-labeled electron-dense particles in their immediate vicinity. (I) Scanning EM of prom1-labeled E10.5 telencephalic ventricular surface; gold particles (18 nm) appear as white dots. Gold-labeled cilia (arrowheads), cup-shaped structures (open arrows), and a strongly immunoreactive protrusion (solid arrow) are indicated. Bars: (A–H) 100 nm; (I) 1 μm.
Figure Legend Snippet: Prom1 is concentrated on cilia of NE cells. Mouse forebrain was subjected to preembedding immunogold labeling for prom1 (10 nm gold) followed by EM analysis of ultrathin plastic sections. (A) A cilium at E8.5 showing weak labeling (arrows). (B–F) Cilia at E10.5 (B and C), E12.5 (D and F), or E13.5 (E) showing different degrees of surface labeling; note the cluster of prom1 at the tip of the cilium in D and E (arrowheads). (G and H) Short cilia at E12.5 with strongly prom1-labeled electron-dense particles in their immediate vicinity. (I) Scanning EM of prom1-labeled E10.5 telencephalic ventricular surface; gold particles (18 nm) appear as white dots. Gold-labeled cilia (arrowheads), cup-shaped structures (open arrows), and a strongly immunoreactive protrusion (solid arrow) are indicated. Bars: (A–H) 100 nm; (I) 1 μm.

Techniques Used: Labeling

24) Product Images from "Extracellular Vesicles Mediate Receptor-Independent Transmission of Novel Tick-Borne Bunyavirus"

Article Title: Extracellular Vesicles Mediate Receptor-Independent Transmission of Novel Tick-Borne Bunyavirus

Journal: Journal of Virology

doi: 10.1128/JVI.02490-15

SFTS virus NSs induces the formation of endosome-like structures. Ultrastructure analyses of SFTS virus NSs-expressing cells (A) and SFTS virus-infected cells (B) show cytoplasmic structures reminiscent of early endosomes (arrows) in ultrathin sections. Immunogold staining of distinct ultrathin sections shows cytoplasmic structures positive for SFTS virus NSs and Rab5 (C) or SFTS virus NSs and LC3B (D). A solid red arrows indicate SFTS virus NSs in panels C and D, while a red triangle (in panel C) and a green triangle (in panel D) indicate the detection of Rab5 and LC3B, respectively. Ultrathin sections of mock-infected cells were also labeled, as indicated above (not shown), to ensure the specificity of antibody. Representatives images are shown.
Figure Legend Snippet: SFTS virus NSs induces the formation of endosome-like structures. Ultrastructure analyses of SFTS virus NSs-expressing cells (A) and SFTS virus-infected cells (B) show cytoplasmic structures reminiscent of early endosomes (arrows) in ultrathin sections. Immunogold staining of distinct ultrathin sections shows cytoplasmic structures positive for SFTS virus NSs and Rab5 (C) or SFTS virus NSs and LC3B (D). A solid red arrows indicate SFTS virus NSs in panels C and D, while a red triangle (in panel C) and a green triangle (in panel D) indicate the detection of Rab5 and LC3B, respectively. Ultrathin sections of mock-infected cells were also labeled, as indicated above (not shown), to ensure the specificity of antibody. Representatives images are shown.

Techniques Used: Expressing, Infection, Staining, Labeling

25) Product Images from "A barley powdery mildew fungus non-autonomous retrotransposon encodes a peptide that supports penetration success on barley"

Article Title: A barley powdery mildew fungus non-autonomous retrotransposon encodes a peptide that supports penetration success on barley

Journal: Journal of Experimental Botany

doi: 10.1093/jxb/ery174

Immunogold labeling of α-ROPIP1 in Bgh -challenged barley leaves. Transmission electron micrographs of ultrathin sections of Bgh -infected barley epidermal cells 3 dai showing gold particles bound to α-ROPIP1. (A, B) Negative control of infected cells treated with a non-specific antibody. Gold particles were absent in the susceptible barley epidermal cell containing intracellular fungal haustorial protrusions (H) and the extracellular Bgh hypha (Hy). (C, D) Gold particles bound to α-ROPIP1 were observed in hyphae, inside a Bgh appressorium (App), the barley epidermal cell wall (CW), and papilla, but were absent from the extracellular space (ES) and the host cell vacuole (V). (E, F) Gold particles were found in the lumen of finger-like Bgh haustorial protrusions inside barley epidermal cells as well as the host cell cytoplasm, but were almost absent from the host cell vacuole (V), the CW, and the ES. Arrowheads in (D) and (F) point to selected gold particles. Scale bars are 1 µm.
Figure Legend Snippet: Immunogold labeling of α-ROPIP1 in Bgh -challenged barley leaves. Transmission electron micrographs of ultrathin sections of Bgh -infected barley epidermal cells 3 dai showing gold particles bound to α-ROPIP1. (A, B) Negative control of infected cells treated with a non-specific antibody. Gold particles were absent in the susceptible barley epidermal cell containing intracellular fungal haustorial protrusions (H) and the extracellular Bgh hypha (Hy). (C, D) Gold particles bound to α-ROPIP1 were observed in hyphae, inside a Bgh appressorium (App), the barley epidermal cell wall (CW), and papilla, but were absent from the extracellular space (ES) and the host cell vacuole (V). (E, F) Gold particles were found in the lumen of finger-like Bgh haustorial protrusions inside barley epidermal cells as well as the host cell cytoplasm, but were almost absent from the host cell vacuole (V), the CW, and the ES. Arrowheads in (D) and (F) point to selected gold particles. Scale bars are 1 µm.

Techniques Used: Labeling, Transmission Assay, Infection, Negative Control

26) Product Images from "Discrete and overlapping functions of peptidoglycan synthases in growth, cell division and virulence of Listeria monocytogenes"

Article Title: Discrete and overlapping functions of peptidoglycan synthases in growth, cell division and virulence of Listeria monocytogenes

Journal: Molecular Microbiology

doi: 10.1111/mmi.12873

Effect of PBP B1, PBP B2 and PBP B3 inactivation on cell morphology. Transmission electron microscopy of ultrathin sections (A, C, E) and scanning electron microscopy (B, D, F) of fixed whole cells of L . monocytogenes strains devoid of class B high molecular weight penicillin-binding proteins. L . monocytogenes strains LMJR27 (I lmo1438 ), LMJR18 (I lmo2039 ) and LMJR41 (Δ lmo0441 ) were grown to mid-logarithmic growth phase in BHI at 37°C and subjected to chemical fixation and subsequent electron microscopy as described in the experimental procedures section. Scale bars: left column 200 nm, right column 2 μm.
Figure Legend Snippet: Effect of PBP B1, PBP B2 and PBP B3 inactivation on cell morphology. Transmission electron microscopy of ultrathin sections (A, C, E) and scanning electron microscopy (B, D, F) of fixed whole cells of L . monocytogenes strains devoid of class B high molecular weight penicillin-binding proteins. L . monocytogenes strains LMJR27 (I lmo1438 ), LMJR18 (I lmo2039 ) and LMJR41 (Δ lmo0441 ) were grown to mid-logarithmic growth phase in BHI at 37°C and subjected to chemical fixation and subsequent electron microscopy as described in the experimental procedures section. Scale bars: left column 200 nm, right column 2 μm.

Techniques Used: Transmission Assay, Electron Microscopy, Molecular Weight, Binding Assay

Effect of PBP A1 and PBP A2 inactivation on cell morphology. Transmission electron microscopy of ultrathin sections (A, C, E, G) and scanning electron microscopy (B, D, F, H) of fixed whole cells of L . monocytogenes strains devoid of class A high molecular weight penicillin-binding proteins. L . monocytogenes strains EGD-e (wt), LMS57 (Δ lmo1892 ), LMS64 (Δ lmo2229 ) and the inducible double mutant strain LMJR30 (I lmo1892 Δ lmo2229 ) were grown to mid-logarithmic growth phase in BHI at 37°C and subjected to chemical fixation and subsequent electron microscopy as described in the experimental procedures section. Scale bars: left column 200 nm, right column 2 μm.
Figure Legend Snippet: Effect of PBP A1 and PBP A2 inactivation on cell morphology. Transmission electron microscopy of ultrathin sections (A, C, E, G) and scanning electron microscopy (B, D, F, H) of fixed whole cells of L . monocytogenes strains devoid of class A high molecular weight penicillin-binding proteins. L . monocytogenes strains EGD-e (wt), LMS57 (Δ lmo1892 ), LMS64 (Δ lmo2229 ) and the inducible double mutant strain LMJR30 (I lmo1892 Δ lmo2229 ) were grown to mid-logarithmic growth phase in BHI at 37°C and subjected to chemical fixation and subsequent electron microscopy as described in the experimental procedures section. Scale bars: left column 200 nm, right column 2 μm.

Techniques Used: Transmission Assay, Electron Microscopy, Molecular Weight, Binding Assay, Mutagenesis

27) Product Images from "Aim24 and MICOS modulate respiratory function, tafazzin-related cardiolipin modification and mitochondrial architecture"

Article Title: Aim24 and MICOS modulate respiratory function, tafazzin-related cardiolipin modification and mitochondrial architecture

Journal: eLife

doi: 10.7554/eLife.01684

Deletion of the AIM24 gene leads to altered mitochondrial morphology and architecture. ( A and B ) Electron microscopy of wild type and AIM24 deletion mutant. Cells were grown in either YPD or YPG and fixed with glutaraldehyde, contrasted with osmium tetroxide and uranylacetate. Ultrathin sections were obtained and contrasted with uranylacetate and lead citrate. ( A ) Wild type. a–c, cells grown on YPD; d–f cells grown on YPG; a and d, cells (scale bar, 1 µm); b, c, e, f, mitochondrial profiles (scale bar, 0.2 µm). ( B ) Δ aim24 . a–i, cells grown on YPG; j–o cells grown on YPD; a, b, j and k, cells (scale bar, 1 µm). c–i and l–o, mitochondrial profiles (scale bars, 0.2 µm). ( C ) Fluorescence microscopy of wild type and Δ aim24 cells expressing mitoGFP. Cells were grown at 30°C on YPD or YPG medium to stationary phase (scale bar, 5 µm). DOI: http://dx.doi.org/10.7554/eLife.01684.010
Figure Legend Snippet: Deletion of the AIM24 gene leads to altered mitochondrial morphology and architecture. ( A and B ) Electron microscopy of wild type and AIM24 deletion mutant. Cells were grown in either YPD or YPG and fixed with glutaraldehyde, contrasted with osmium tetroxide and uranylacetate. Ultrathin sections were obtained and contrasted with uranylacetate and lead citrate. ( A ) Wild type. a–c, cells grown on YPD; d–f cells grown on YPG; a and d, cells (scale bar, 1 µm); b, c, e, f, mitochondrial profiles (scale bar, 0.2 µm). ( B ) Δ aim24 . a–i, cells grown on YPG; j–o cells grown on YPD; a, b, j and k, cells (scale bar, 1 µm). c–i and l–o, mitochondrial profiles (scale bars, 0.2 µm). ( C ) Fluorescence microscopy of wild type and Δ aim24 cells expressing mitoGFP. Cells were grown at 30°C on YPD or YPG medium to stationary phase (scale bar, 5 µm). DOI: http://dx.doi.org/10.7554/eLife.01684.010

Techniques Used: Electron Microscopy, Mutagenesis, Fluorescence, Microscopy, Expressing

28) Product Images from "Interactions between colon cancer cells and hepatocytes in rats in relation to metastasis"

Article Title: Interactions between colon cancer cells and hepatocytes in rats in relation to metastasis

Journal: Journal of Cellular and Molecular Medicine

doi: 10.1111/j.1582-4934.2008.00242.x

Electron micrograph (A) and electron tomographic reconstruction (B) of the intercellular space between a cultured colon cancer cell and a suspended hepatocyte after 1 hr coculture showing electron-dense molecular contacts. The reconstructed area (0.45 μm x 0.28 μm) is indicated. Thickness of the ultrathin section was 120 nm. Bar = 1 μm.
Figure Legend Snippet: Electron micrograph (A) and electron tomographic reconstruction (B) of the intercellular space between a cultured colon cancer cell and a suspended hepatocyte after 1 hr coculture showing electron-dense molecular contacts. The reconstructed area (0.45 μm x 0.28 μm) is indicated. Thickness of the ultrathin section was 120 nm. Bar = 1 μm.

Techniques Used: Cell Culture

29) Product Images from "Agitation Modules: Flexible Means to Accelerate Automated Freeze Substitution"

Article Title: Agitation Modules: Flexible Means to Accelerate Automated Freeze Substitution

Journal: Journal of Histochemistry and Cytochemistry

doi: 10.1369/0022155418786698

Immunogold labeling of Lowicryl HM20 sections of HP-frozen, low-temperature embedded immature anthers of Arabidopsis thaliana generated by accelerated FS under agitation. (A) Overview image showing a middle layer cell from an anther surrounded by anti-xyloglucan labeled cell wall (cw). (B) Aggregation of intracellular vesicle-like xyloglucan-positive structures. (C) Overview image showing anti-α-tubulin labeling close to the plasma membrane. (D) Details of immunolabeled cortical microtubules (arrows) aligned almost in parallel to the section plane. Typical for postembedding labeling, the gold-conjugated antibodies are concentrated at regions where the MTs meet the surface of the ultrathin section. Scale bars = 1 µm in A and C; 300 nm in B and D. Abbreviations: n, nucleus; v, vacuole; chl, chloroplast; HP, high-pressure; FS, freeze substitution; MT, microtubule.
Figure Legend Snippet: Immunogold labeling of Lowicryl HM20 sections of HP-frozen, low-temperature embedded immature anthers of Arabidopsis thaliana generated by accelerated FS under agitation. (A) Overview image showing a middle layer cell from an anther surrounded by anti-xyloglucan labeled cell wall (cw). (B) Aggregation of intracellular vesicle-like xyloglucan-positive structures. (C) Overview image showing anti-α-tubulin labeling close to the plasma membrane. (D) Details of immunolabeled cortical microtubules (arrows) aligned almost in parallel to the section plane. Typical for postembedding labeling, the gold-conjugated antibodies are concentrated at regions where the MTs meet the surface of the ultrathin section. Scale bars = 1 µm in A and C; 300 nm in B and D. Abbreviations: n, nucleus; v, vacuole; chl, chloroplast; HP, high-pressure; FS, freeze substitution; MT, microtubule.

Techniques Used: Labeling, Generated, Immunolabeling

30) Product Images from "The PIKfyve complex regulates the early melanosome homeostasis required for physiological amyloid formation"

Article Title: The PIKfyve complex regulates the early melanosome homeostasis required for physiological amyloid formation

Journal: Journal of Cell Science

doi: 10.1242/jcs.229500

PMEL fragments accumulate in enlarged EEA1 compartments. (A,B) EM analysis of Epon-embedded MNT-1 cells treated for 2 h with the PIKfyve inhibitor YM201636 (A) or knocked down for VAC14, FIG4 and PIKfyve (B). Upper panels show low-magnification overviews of unpigmented and pigmented melanosomes, and lower panels show unpigmented melanosomes at higher magnification. Arrowheads point towards enlarged stage I melanosomes and arrows highlight aberrant unpigmented melanosomes containing unstructured aggregates present in VAC14, FIG4 and PIKfyve knockdown cells. I–IV indicates the melanosome stage. Scale bars: 500 nm (upper panels), 200 nm (lower panels). (C) Quantification of stage I melanosome size. (D) Quantification of unpigmented melanosomes grouped into stage I, stage II and aberrant melanosomes. (E) Quantification of pigmented and unpigmented melanosomes expressed as a percentage of the total number of melanosomes. (F) EM analysis of Epon-embedded human primary melanocytes treated with control siRNAs or siRNAs against VAC14, FIG4 and PIKfyve. Right panels show magnifications of stage I melanosomes. I–IV indicates the melanosome stage. Scale bars: 1 µm (left panels), and 500 nm (right panels). (G) Ultrathin cryosections of MNT-1 cells treated for 2 h with 1.6 µM YM201636 or DMSO were immunogold-labeled using anti-PMEL-N antibody followed by protein-A conjugated to 15 nm diameter gold. Arrows indicate clathrin coats. Scale bars: 200 nm. (H) Quantification of the percentage of gold particles on ILVs expressed as a percentage of the total number of gold particles. (I) Quantification of the number of gold particles on ILVs per compartment. Means±s.e.m. shown for n ≥3 in all panels. * P
Figure Legend Snippet: PMEL fragments accumulate in enlarged EEA1 compartments. (A,B) EM analysis of Epon-embedded MNT-1 cells treated for 2 h with the PIKfyve inhibitor YM201636 (A) or knocked down for VAC14, FIG4 and PIKfyve (B). Upper panels show low-magnification overviews of unpigmented and pigmented melanosomes, and lower panels show unpigmented melanosomes at higher magnification. Arrowheads point towards enlarged stage I melanosomes and arrows highlight aberrant unpigmented melanosomes containing unstructured aggregates present in VAC14, FIG4 and PIKfyve knockdown cells. I–IV indicates the melanosome stage. Scale bars: 500 nm (upper panels), 200 nm (lower panels). (C) Quantification of stage I melanosome size. (D) Quantification of unpigmented melanosomes grouped into stage I, stage II and aberrant melanosomes. (E) Quantification of pigmented and unpigmented melanosomes expressed as a percentage of the total number of melanosomes. (F) EM analysis of Epon-embedded human primary melanocytes treated with control siRNAs or siRNAs against VAC14, FIG4 and PIKfyve. Right panels show magnifications of stage I melanosomes. I–IV indicates the melanosome stage. Scale bars: 1 µm (left panels), and 500 nm (right panels). (G) Ultrathin cryosections of MNT-1 cells treated for 2 h with 1.6 µM YM201636 or DMSO were immunogold-labeled using anti-PMEL-N antibody followed by protein-A conjugated to 15 nm diameter gold. Arrows indicate clathrin coats. Scale bars: 200 nm. (H) Quantification of the percentage of gold particles on ILVs expressed as a percentage of the total number of gold particles. (I) Quantification of the number of gold particles on ILVs per compartment. Means±s.e.m. shown for n ≥3 in all panels. * P

Techniques Used: Labeling

31) Product Images from "Dynamic compartment specific changes in glutathione and ascorbate levels in Arabidopsis plants exposed to different light intensities"

Article Title: Dynamic compartment specific changes in glutathione and ascorbate levels in Arabidopsis plants exposed to different light intensities

Journal: BMC Plant Biology

doi: 10.1186/1471-2229-13-104

Areas of internal chloroplast structures after the exposure to different light intensities for 14 d. Area (in μm 2 ) and percentage of areas of internal chloroplast structures detected by TEM on a longitudinal ultrathin section within the mesophyll of leaves from the wildtype (Col-0), the pad2-1 and the vtc2-1 mutants after the exposure to different light intensities for 14 d. Data are means with standard errors. Significant differences were calculated within one line of plants between control conditions (exposure to 150 μmol m -2 s -1 ) and the same line exposed to the other light intensities by using the Mann Whitney U-test; *, ** and ***, respectively, indicate significance at the 0.05, 0.01 and 0.001 levels of confidence. n > 100 chloroplasts per treatment.
Figure Legend Snippet: Areas of internal chloroplast structures after the exposure to different light intensities for 14 d. Area (in μm 2 ) and percentage of areas of internal chloroplast structures detected by TEM on a longitudinal ultrathin section within the mesophyll of leaves from the wildtype (Col-0), the pad2-1 and the vtc2-1 mutants after the exposure to different light intensities for 14 d. Data are means with standard errors. Significant differences were calculated within one line of plants between control conditions (exposure to 150 μmol m -2 s -1 ) and the same line exposed to the other light intensities by using the Mann Whitney U-test; *, ** and ***, respectively, indicate significance at the 0.05, 0.01 and 0.001 levels of confidence. n > 100 chloroplasts per treatment.

Techniques Used: Transmission Electron Microscopy, MANN-WHITNEY

32) Product Images from "Receptor-Mediated Endocytosis of Lysozyme in Renal Proximal Tubules of the Frog Rana Temporaria"

Article Title: Receptor-Mediated Endocytosis of Lysozyme in Renal Proximal Tubules of the Frog Rana Temporaria

Journal: European Journal of Histochemistry : EJH

doi: 10.4081/ejh.2015.2482

Electron micrographs of the proximal tubule cells from lysozyme-injected frogs. A) Well-preserved ultrastructure of endocytic apparatus epithelial cells after lysozyme injection. B-G) Post-embedding immunogold labeling of ultrathin LR-White sections from the frog kidneys. B-D) Immunogold staining of lysozyme 10 min (B, C) and 30 min (D) after injection; label is detected over the brush border, in apical vesicles, endosomes and vesicles with a semi-dense content (D). E) Localization of clathrin in microvilli and apical vesicles in the tubular cells 10 min after injection. F) Immunolabeling of megalin in the tubular cells of lysozyme-treated frogs 10 min after injection; localization of this receptor in apical endocytic compartment is shown. G) The same for cubilin. Arrow shows the label in apical coated pits; asterisk shows immunogold particles in vesicles. E, endosome; M, mitochondria; MV, microvilli; V, apical vesicle. Scale bars, 1 µm.
Figure Legend Snippet: Electron micrographs of the proximal tubule cells from lysozyme-injected frogs. A) Well-preserved ultrastructure of endocytic apparatus epithelial cells after lysozyme injection. B-G) Post-embedding immunogold labeling of ultrathin LR-White sections from the frog kidneys. B-D) Immunogold staining of lysozyme 10 min (B, C) and 30 min (D) after injection; label is detected over the brush border, in apical vesicles, endosomes and vesicles with a semi-dense content (D). E) Localization of clathrin in microvilli and apical vesicles in the tubular cells 10 min after injection. F) Immunolabeling of megalin in the tubular cells of lysozyme-treated frogs 10 min after injection; localization of this receptor in apical endocytic compartment is shown. G) The same for cubilin. Arrow shows the label in apical coated pits; asterisk shows immunogold particles in vesicles. E, endosome; M, mitochondria; MV, microvilli; V, apical vesicle. Scale bars, 1 µm.

Techniques Used: Injection, Labeling, Staining, Immunolabeling

33) Product Images from "Early and Late Pathogenic Events of Newborn Mice Encephalitis Experimentally Induced by Itacaiunas and Curionópolis Bracorhabdoviruses Infection"

Article Title: Early and Late Pathogenic Events of Newborn Mice Encephalitis Experimentally Induced by Itacaiunas and Curionópolis Bracorhabdoviruses Infection

Journal: PLoS ONE

doi: 10.1371/journal.pone.0001733

Transmission electron photomicrographs of ultrathin sections obtained from primary neuronal cultures. Control cultures of normal cells (A) and after Itacaiunas (B) and Curionopolis (C, D) infection at 4 and 5 days post-inoculation, respectively. Apoptotic cells (C) and virus budding (rectangle and arrows) (C, D) after Curionopolis infection. Itacaiunas virus particles (arrows) in culture at 5 days post-inoculation (B). N: nucleus; AN: apoptotic nucleus.
Figure Legend Snippet: Transmission electron photomicrographs of ultrathin sections obtained from primary neuronal cultures. Control cultures of normal cells (A) and after Itacaiunas (B) and Curionopolis (C, D) infection at 4 and 5 days post-inoculation, respectively. Apoptotic cells (C) and virus budding (rectangle and arrows) (C, D) after Curionopolis infection. Itacaiunas virus particles (arrows) in culture at 5 days post-inoculation (B). N: nucleus; AN: apoptotic nucleus.

Techniques Used: Transmission Assay, Infection

Transmission electron photomicrographs of ultrathin sections obtained from control (A) and mouse brain infected intracerebrally with Curionopolis for 36 (B,C), 60 (D) and 96 h (E), and with Itacaiunas for 24 (F), 60 (G), 72 (H), 96 (I) and 108 h (J). Normal tissue with intact neuronal soma and appendages (A); viral particles (arrow), interstitial edema (stars) and cellular rarefaction (lozenge) are seen 36 h post-inoculation (p.i.) (B, C); necrotic cells were observed at 60 h p.i. (D); intense perivascular edema (stars), hyperplastic endotheliocytes and reduced vessel luminal area (E); well-preserved brain parenchyma and vessels at 24 h p.i. (F); viral particles, endotheliocyte hyperplasia, and mild interstitial edema (stars) at 60 h p.i. (G); membrane viral budding in rich polyribosomes oligodendrocyte-like cell at 72 h p.i. (H); brain parenchyma at 96 h p.i. presenting a large number of viral particles (I); apoptotic features were more marked at 108 h p.i. (J). AC = apoptotic cell, M = mitochondria, OL = oligodendrocyte, EC = endothelial cells, VL = vascular lumen, N = cell nucleus, NC = necrotic cells.
Figure Legend Snippet: Transmission electron photomicrographs of ultrathin sections obtained from control (A) and mouse brain infected intracerebrally with Curionopolis for 36 (B,C), 60 (D) and 96 h (E), and with Itacaiunas for 24 (F), 60 (G), 72 (H), 96 (I) and 108 h (J). Normal tissue with intact neuronal soma and appendages (A); viral particles (arrow), interstitial edema (stars) and cellular rarefaction (lozenge) are seen 36 h post-inoculation (p.i.) (B, C); necrotic cells were observed at 60 h p.i. (D); intense perivascular edema (stars), hyperplastic endotheliocytes and reduced vessel luminal area (E); well-preserved brain parenchyma and vessels at 24 h p.i. (F); viral particles, endotheliocyte hyperplasia, and mild interstitial edema (stars) at 60 h p.i. (G); membrane viral budding in rich polyribosomes oligodendrocyte-like cell at 72 h p.i. (H); brain parenchyma at 96 h p.i. presenting a large number of viral particles (I); apoptotic features were more marked at 108 h p.i. (J). AC = apoptotic cell, M = mitochondria, OL = oligodendrocyte, EC = endothelial cells, VL = vascular lumen, N = cell nucleus, NC = necrotic cells.

Techniques Used: Transmission Assay, Infection

34) Product Images from "Autophagy regulates UBC9 levels during viral-mediated tumorigenesis"

Article Title: Autophagy regulates UBC9 levels during viral-mediated tumorigenesis

Journal: PLoS Pathogens

doi: 10.1371/journal.ppat.1006262

HPV16 E6/E7 inhibit late autophagy. (A) Immunolocalization of LC3 in ultrathin sections of HKs transduced with empty or HPV16 E6/E7 vectors. Original view (left) and higher magnification (right) of boxed regions. Nanogold particles are selectively enriched in autophagic structures highlighted by arrows. Scale bar = 1 μm. (B) Top: Confocal microscopy images of empty or HPV16 E6/E7 transduced HKs immunostained with antibody against LC3 (red). Nuclei were stained with DAPI (blue). Scale bar = 5 μm. Bottom: Quantification of endogenous LC3 dot total area. Data are expressed as number of LC3 dots. Bars represent means ± SEM of n = 50 fields of three different biological replicates. ***P
Figure Legend Snippet: HPV16 E6/E7 inhibit late autophagy. (A) Immunolocalization of LC3 in ultrathin sections of HKs transduced with empty or HPV16 E6/E7 vectors. Original view (left) and higher magnification (right) of boxed regions. Nanogold particles are selectively enriched in autophagic structures highlighted by arrows. Scale bar = 1 μm. (B) Top: Confocal microscopy images of empty or HPV16 E6/E7 transduced HKs immunostained with antibody against LC3 (red). Nuclei were stained with DAPI (blue). Scale bar = 5 μm. Bottom: Quantification of endogenous LC3 dot total area. Data are expressed as number of LC3 dots. Bars represent means ± SEM of n = 50 fields of three different biological replicates. ***P

Techniques Used: Transduction, Confocal Microscopy, Staining

UBC9 is degraded by autophagy in epithelial cells. (A) Immunolocalization of UBC9 in ultrathin sections of HKs transduced with empty or HPV16 E6/E7 vectors. Original view (left) and higher magnification (right) of boxed regions. Gold particles are selectively enriched in autophagic structures highlighted by arrows. Scale bar = 1 μm. (B) Top: Representative WB of HaCaT cells treated with the indicated autophagic activators (left) and inhibitors (right). Activation of autophagy was monitored by the conversion of LC3 (LC3-I) to the lipidated LC3 (LC3-II) form, a marker of autophagosome production induced by autophagic stimuli [ 41 ]. LC3-II accumulation was used to verify autophagic impairment [ 41 ]. Bottom: normalized UBC9 expression. Data are expressed as fold over untreated cells. Bars represent means ± SEM of n = 4 different biological replicates. ns: not significant (Kruskal–Wallis one-way ANOVA with Dunn’s post hoc test) compared to vehicle control groups. ( C) Representative WB analysis of U-2 OS or MCF7 cells treated with chloroquine. n = 3 different biological replicates. (D) Representative WB analysis of MCF7 cells transduced with scramble or ATG5 shRNA. The observed ATG5 band represents the ATG5-ATG12 conjugated form. LC3-I accumulation is reported to evidence autophagic deficiencies promoted by shATG5. n = 3 replicates of a single transduction.
Figure Legend Snippet: UBC9 is degraded by autophagy in epithelial cells. (A) Immunolocalization of UBC9 in ultrathin sections of HKs transduced with empty or HPV16 E6/E7 vectors. Original view (left) and higher magnification (right) of boxed regions. Gold particles are selectively enriched in autophagic structures highlighted by arrows. Scale bar = 1 μm. (B) Top: Representative WB of HaCaT cells treated with the indicated autophagic activators (left) and inhibitors (right). Activation of autophagy was monitored by the conversion of LC3 (LC3-I) to the lipidated LC3 (LC3-II) form, a marker of autophagosome production induced by autophagic stimuli [ 41 ]. LC3-II accumulation was used to verify autophagic impairment [ 41 ]. Bottom: normalized UBC9 expression. Data are expressed as fold over untreated cells. Bars represent means ± SEM of n = 4 different biological replicates. ns: not significant (Kruskal–Wallis one-way ANOVA with Dunn’s post hoc test) compared to vehicle control groups. ( C) Representative WB analysis of U-2 OS or MCF7 cells treated with chloroquine. n = 3 different biological replicates. (D) Representative WB analysis of MCF7 cells transduced with scramble or ATG5 shRNA. The observed ATG5 band represents the ATG5-ATG12 conjugated form. LC3-I accumulation is reported to evidence autophagic deficiencies promoted by shATG5. n = 3 replicates of a single transduction.

Techniques Used: Transduction, Western Blot, Activation Assay, Marker, Expressing, shRNA

35) Product Images from "Non-Covalent Associates of siRNAs and AuNPs Enveloped with Lipid Layer and Doped with Amphiphilic Peptide for Efficient siRNA Delivery"

Article Title: Non-Covalent Associates of siRNAs and AuNPs Enveloped with Lipid Layer and Doped with Amphiphilic Peptide for Efficient siRNA Delivery

Journal: International Journal of Molecular Sciences

doi: 10.3390/ijms19072096

Visualization of AuNPs (serve as a marker of siRNA-Au-LM-P”) in endosome-lysosome system of HEK Phoenix cells 72 h after transfection with siRNA-Au-LM2-P’’: ( a ) late endosome; ( b ) lysosome; ( c ) autophagosome, AuNPs with low contrast are shown the insert; and ( d , e ) different lysosome structures. Arrows show membrane envelope of endosome-lysosome structures. Length of scale bars corresponds to 200 nm. TEM, ultrathin sections.
Figure Legend Snippet: Visualization of AuNPs (serve as a marker of siRNA-Au-LM-P”) in endosome-lysosome system of HEK Phoenix cells 72 h after transfection with siRNA-Au-LM2-P’’: ( a ) late endosome; ( b ) lysosome; ( c ) autophagosome, AuNPs with low contrast are shown the insert; and ( d , e ) different lysosome structures. Arrows show membrane envelope of endosome-lysosome structures. Length of scale bars corresponds to 200 nm. TEM, ultrathin sections.

Techniques Used: Marker, Transfection, Transmission Electron Microscopy

36) Product Images from "Human Herpesvirus-6 Induces MVB Formation, and Virus Egress Occurs by an Exosomal Release Pathway"

Article Title: Human Herpesvirus-6 Induces MVB Formation, and Virus Egress Occurs by an Exosomal Release Pathway

Journal: Traffic (Copenhagen, Denmark)

doi: 10.1111/j.1600-0854.2008.00796.x

Formation of MVBs containing enveloped virions in the cytoplasm of HHV6-infected cells Ultrathin sections of Epon-embedded cells. A) A massive accumulation of MVBs in the perinuclear region of an infected cell. The MVBs often contain enveloped virions (arrows). g, Golgi apparatus. B) Examples of MVBs with numerous intact enveloped virions and internal vesicles. C) Small vacuoles (arrows) containing virions adjacent to Golgi apparatus (g) and larger MVB-like vacuoles containing virions and internal small vesicles (arrowheads). Scale bars: 0.5 μm.
Figure Legend Snippet: Formation of MVBs containing enveloped virions in the cytoplasm of HHV6-infected cells Ultrathin sections of Epon-embedded cells. A) A massive accumulation of MVBs in the perinuclear region of an infected cell. The MVBs often contain enveloped virions (arrows). g, Golgi apparatus. B) Examples of MVBs with numerous intact enveloped virions and internal vesicles. C) Small vacuoles (arrows) containing virions adjacent to Golgi apparatus (g) and larger MVB-like vacuoles containing virions and internal small vesicles (arrowheads). Scale bars: 0.5 μm.

Techniques Used: Infection

Immunocytochemical detection of AP-1, CD63, gM and gB around the TGN in HHV6A-infected cells Immunogold labeling showing AP-1 (A and B) (5 nm), TGN-46 (C) (5 nm), CD63 (D–F) (10 nm), gM (G and H) (10 nm) or gB (I) (10 nm) or double labeling indicating gM (15 nm) and gB (10 nm) (J) in HHV6A-infected cells on ultrathin cryosections. A and B) Positive signals for AP-1 were localized to vacuoles closely associated with the TGN (asterisks). Note that tubulo-vacuoles containing or incorporating virions (v) (arrowheads) were also immunopositive for AP-1 (arrows). C) Positive signals for TGN-46 are localized to small vacuoles (asterisks) and (tubulo-)vacuoles containing virions (v) (arrowheads) but not in an MVB. D–F) Positive CD63 signals were localized not only to MVBs but were also on (tubulo-)vacuolar structures associated with the TGN (asterisks). Vacuoles that were labeled with immunogold particles for CD63 frequently had incorporated virions (v) (arrowheads). G–I) Positive gM (G and H) or gB (I) signals were seen on tubulo-vacuolar structures (arrows) or vacuoles (asterisks) closely associated with the TGN. Note that a vacuole incorporating a virion (v) (an arrowhead) was heavily labeled with immunogold particles for gB (I). gB signals were also localized to MVBs. J) Vacuoles closely associated with the TGN were often co-labeled with gM (15 nm) and gB (10 nm) (asterisks). g, Golgi complex. Scale bars: 0.5 μm.
Figure Legend Snippet: Immunocytochemical detection of AP-1, CD63, gM and gB around the TGN in HHV6A-infected cells Immunogold labeling showing AP-1 (A and B) (5 nm), TGN-46 (C) (5 nm), CD63 (D–F) (10 nm), gM (G and H) (10 nm) or gB (I) (10 nm) or double labeling indicating gM (15 nm) and gB (10 nm) (J) in HHV6A-infected cells on ultrathin cryosections. A and B) Positive signals for AP-1 were localized to vacuoles closely associated with the TGN (asterisks). Note that tubulo-vacuoles containing or incorporating virions (v) (arrowheads) were also immunopositive for AP-1 (arrows). C) Positive signals for TGN-46 are localized to small vacuoles (asterisks) and (tubulo-)vacuoles containing virions (v) (arrowheads) but not in an MVB. D–F) Positive CD63 signals were localized not only to MVBs but were also on (tubulo-)vacuolar structures associated with the TGN (asterisks). Vacuoles that were labeled with immunogold particles for CD63 frequently had incorporated virions (v) (arrowheads). G–I) Positive gM (G and H) or gB (I) signals were seen on tubulo-vacuolar structures (arrows) or vacuoles (asterisks) closely associated with the TGN. Note that a vacuole incorporating a virion (v) (an arrowhead) was heavily labeled with immunogold particles for gB (I). gB signals were also localized to MVBs. J) Vacuoles closely associated with the TGN were often co-labeled with gM (15 nm) and gB (10 nm) (asterisks). g, Golgi complex. Scale bars: 0.5 μm.

Techniques Used: Infection, Labeling

Accumulation of gB, gM and CD63 in MVBs Immunogold labeling on ultrathin cryosections of HHV6A-infected cells showing gM (A) (10 nm), gB (B) or CD63 (D) (10 nm) or double labeling indicating gM (15 nm) and gB (10 nm) (C) or CD63 (5 nm) and gM (10 nm) (E) or gB (10 nm) (F). A–C) Immunogold particles indicating gM and/or gB were localized to both the internal vesicles of MVBs (asterisks) and the virions (v) within them. Note that extracellular virions (arrows), smaller vacuoles that contained only virions (arrowheads) and the Golgi apparatus (g) were also immunopositive for gM and gB. D) CD63 was localized to the internal vesicles of the MVBs (asterisk) and the virions (v) within them. E and F) Immunogold particles indicating gM (E) or gB (F) with CD63 were colocalized to the internal vesicles of the MVBs (asterisks) and the virions (v) within them. Scale bars: 0.5 μm.
Figure Legend Snippet: Accumulation of gB, gM and CD63 in MVBs Immunogold labeling on ultrathin cryosections of HHV6A-infected cells showing gM (A) (10 nm), gB (B) or CD63 (D) (10 nm) or double labeling indicating gM (15 nm) and gB (10 nm) (C) or CD63 (5 nm) and gM (10 nm) (E) or gB (10 nm) (F). A–C) Immunogold particles indicating gM and/or gB were localized to both the internal vesicles of MVBs (asterisks) and the virions (v) within them. Note that extracellular virions (arrows), smaller vacuoles that contained only virions (arrowheads) and the Golgi apparatus (g) were also immunopositive for gM and gB. D) CD63 was localized to the internal vesicles of the MVBs (asterisk) and the virions (v) within them. E and F) Immunogold particles indicating gM (E) or gB (F) with CD63 were colocalized to the internal vesicles of the MVBs (asterisks) and the virions (v) within them. Scale bars: 0.5 μm.

Techniques Used: Labeling, Infection

Exosomes contribute to the release of enveloped virions A–C) Ultrathin sections of Epon-embedded cells. MVBs were exocytosed from the cell surface, and numerous virions along with small internal vesicles were released from the cells. Released virions and exosome-like small internal vesicles were detected in the vicinity of the cell membrane (B and C). D) Double immunogold labeling of gB (5 nm) and CD63 (10 nm) in HHV6A-infected cells on ultrathin cryosections. Some virions (v) and exosomes (arrows) were positive for both gB and CD63. PM, plasma membrane. E–H) Exosome fractions containing virions were collected by sucrose density gradient from the culture medium of HHV-6A-infected T cells and were analyzed by immunoblotting and EM. E and F) Immunoblot analysis with anti-gB (E) or anti-CD63 (F) antibodies of the sucrose density gradient fractions. The same volume of the gradient fraction was used for all blots. Densities of the fractions are listed at the bottom in grams/milliliters. G and H) Immunogold labeling of gB (G) and CD63 (H) in whole-mount virions and exosomes from fraction 9 in panels (E) or (F). Gold particles showing gB (G) and CD63 (H) labeling of exosomes (arrowheads) and virions (v). Insets in (G and H) showed immunogold labeling of gB and CD63 in purified virions, respectively. Gold particles indicating gB [an inset in (G)] and CD63 [an inset in (H)] were detected in purified virions (v). Scale bars: 1 μm (A–C), 0.5 μm (D), 100 nm [(G and H) and insets in (G and H)].
Figure Legend Snippet: Exosomes contribute to the release of enveloped virions A–C) Ultrathin sections of Epon-embedded cells. MVBs were exocytosed from the cell surface, and numerous virions along with small internal vesicles were released from the cells. Released virions and exosome-like small internal vesicles were detected in the vicinity of the cell membrane (B and C). D) Double immunogold labeling of gB (5 nm) and CD63 (10 nm) in HHV6A-infected cells on ultrathin cryosections. Some virions (v) and exosomes (arrows) were positive for both gB and CD63. PM, plasma membrane. E–H) Exosome fractions containing virions were collected by sucrose density gradient from the culture medium of HHV-6A-infected T cells and were analyzed by immunoblotting and EM. E and F) Immunoblot analysis with anti-gB (E) or anti-CD63 (F) antibodies of the sucrose density gradient fractions. The same volume of the gradient fraction was used for all blots. Densities of the fractions are listed at the bottom in grams/milliliters. G and H) Immunogold labeling of gB (G) and CD63 (H) in whole-mount virions and exosomes from fraction 9 in panels (E) or (F). Gold particles showing gB (G) and CD63 (H) labeling of exosomes (arrowheads) and virions (v). Insets in (G and H) showed immunogold labeling of gB and CD63 in purified virions, respectively. Gold particles indicating gB [an inset in (G)] and CD63 [an inset in (H)] were detected in purified virions (v). Scale bars: 1 μm (A–C), 0.5 μm (D), 100 nm [(G and H) and insets in (G and H)].

Techniques Used: Labeling, Infection, Purification

Accumulation of tegument-like materials along the cytosolic face of the TGN-derived vacuoles Ultrathin sections of Epon-embedded cells. A) Numerous vacuoles were observed around the Golgi apparatus (g), and their membrane domains were modified with electron-dense tegument-like materials (arrowheads). Note the microtubule-organizing center (boxed area and the inset). B) Large tubulo-vacuoles (asterisk) were located in or near the TGN (tgn) and often contained enveloped virions, while clathrin-coated vesicles (arrow) budding from such tubulo-vacuoles were observed. Arrowheads indicate electron-dense materials resembling tegument. Moreover, inwardly budding profiles of the membrane of such tubulo-vacuoles (open arrow) with internal vesicles (open arrowhead) were observed. C and D) Examples of vacuoles with multiple membrane domains decorated with electron-dense tegument-like materials (arrowheads). E) An example of the vacuoles with budding clathrin-coated vesicles and multiple membrane domains with electron-dense tegument-like materials (arrow). F) L-particle (arrow) was found near plasma membrane. Scale bars: 0.3 μm.
Figure Legend Snippet: Accumulation of tegument-like materials along the cytosolic face of the TGN-derived vacuoles Ultrathin sections of Epon-embedded cells. A) Numerous vacuoles were observed around the Golgi apparatus (g), and their membrane domains were modified with electron-dense tegument-like materials (arrowheads). Note the microtubule-organizing center (boxed area and the inset). B) Large tubulo-vacuoles (asterisk) were located in or near the TGN (tgn) and often contained enveloped virions, while clathrin-coated vesicles (arrow) budding from such tubulo-vacuoles were observed. Arrowheads indicate electron-dense materials resembling tegument. Moreover, inwardly budding profiles of the membrane of such tubulo-vacuoles (open arrow) with internal vesicles (open arrowhead) were observed. C and D) Examples of vacuoles with multiple membrane domains decorated with electron-dense tegument-like materials (arrowheads). E) An example of the vacuoles with budding clathrin-coated vesicles and multiple membrane domains with electron-dense tegument-like materials (arrow). F) L-particle (arrow) was found near plasma membrane. Scale bars: 0.3 μm.

Techniques Used: Derivative Assay, Modification

Intracellular distribution of the Golgi apparatus, MVBs and AL in an HHV6A-infected cell Ultrathin sections of Epon-embedded cells. A cluster of Golgi apparatus together with tubulo-vacuolar structures [marked by green color in (A) and Golgi in (B)] was located in the center of surrounding MVBs [marked by red color in (A) and MVB in (B)]. AL [blue in (A) and marked ‘AL’ in (B)] were observed apart from the Golgi apparatus and MVBs. Scale bars: 3 μm.
Figure Legend Snippet: Intracellular distribution of the Golgi apparatus, MVBs and AL in an HHV6A-infected cell Ultrathin sections of Epon-embedded cells. A cluster of Golgi apparatus together with tubulo-vacuolar structures [marked by green color in (A) and Golgi in (B)] was located in the center of surrounding MVBs [marked by red color in (A) and MVB in (B)]. AL [blue in (A) and marked ‘AL’ in (B)] were observed apart from the Golgi apparatus and MVBs. Scale bars: 3 μm.

Techniques Used: Infection

Electron microscopic detection of secondary envelopment Ultrathin sections of Epon-embedded cells. A) Enveloped virions were detected in vacuoles near the Golgi complex (g), and such vacuoles possessed clathrin-coated membrane domains that frequently formed a bud. B–F) Numerous capsids in the cytoplasm close to very dense membrane domains of vacuoles that resembled secondary envelope (B and F) were enwrapped by the membrane to form inward blebs or buds into the vacuolar space [asterisks in (B and F)] and eventually appeared as enveloped virions within the space (D–F). Small arrows in (B–D) indicate clathrin-coated membrane domains. Arrowheads indicate clathrin-coated vesicles budding from the vacuoles containing enveloped virions (E and F). An arrow in (E) indicates part of a polygonal network showing the ‘honeycomb pattern’ formed by clathrin. G and H) Large tubulo-vacuoles (asterisks) were located in or near the TGN (tgn), and they often contained enveloped virions, and clathrin-coated vesicles (arrowheads) budding from such tubulo-vacuoles were observed. Moreover, inwardly budding profiles of such tubulo-vacuoles [open arrows in (G)] and internal vesicles [open arrowhead in (G)] within them were also observed. Scale bars: 0.3 μm.
Figure Legend Snippet: Electron microscopic detection of secondary envelopment Ultrathin sections of Epon-embedded cells. A) Enveloped virions were detected in vacuoles near the Golgi complex (g), and such vacuoles possessed clathrin-coated membrane domains that frequently formed a bud. B–F) Numerous capsids in the cytoplasm close to very dense membrane domains of vacuoles that resembled secondary envelope (B and F) were enwrapped by the membrane to form inward blebs or buds into the vacuolar space [asterisks in (B and F)] and eventually appeared as enveloped virions within the space (D–F). Small arrows in (B–D) indicate clathrin-coated membrane domains. Arrowheads indicate clathrin-coated vesicles budding from the vacuoles containing enveloped virions (E and F). An arrow in (E) indicates part of a polygonal network showing the ‘honeycomb pattern’ formed by clathrin. G and H) Large tubulo-vacuoles (asterisks) were located in or near the TGN (tgn), and they often contained enveloped virions, and clathrin-coated vesicles (arrowheads) budding from such tubulo-vacuoles were observed. Moreover, inwardly budding profiles of such tubulo-vacuoles [open arrows in (G)] and internal vesicles [open arrowhead in (G)] within them were also observed. Scale bars: 0.3 μm.

Techniques Used:

Electron micrographs showing the primary envelopment and nuclear egress of HHV-6 capsids and the formation of AL in HHV-6-infected cells A–I) Ultrathin sections of Epon-embedded cells. J–N) Ultrathin cryosections. A–E) Translocation of capsids from the nucleus (n) to the cytoplasm (c). Intranuclear capsids (A), budding of a capsid from the inner nuclear membrane (primary envelopment) (B), a primary enveloped virion in the cisternal space of the nuclear membrane (C), fusion of primary envelope with the outer nuclear membrane (D) and translocation of the capsids into the cytosol (E). F–I) AL were clearly detected near the nuclear membrane of an infected cell (F). The boxed area in (F) is shown in the inset. The outer membrane of the nuclear envelope was closely associated with the membrane of AL [arrowheads in (F) and the inset in (F)]. Membrane association was also seen between AL and the rough endoplasmic reticulum [arrows in (F and G)]. Note that vesicular or tubular membranes that enwrapped or surrounded virions [arrowheads in (H and I): the boxed area in (H)] were distinct from the membranes of AL cisternae [arrows in (H and I)]. Immunolabeling of gM (J and K) or gB (L) (10 nm) and double immunolabeling of gM (15 nm) and gB (10 nm) (M) on ultrathin cryosections. Gold particles indicating gM- and/or gB-labeled virions (v) and small vesicles located in tubulo-vacuoles (asterisks) in the Golgi complex (g) and MVB but not in AL in the cells. Arrowheads indicate gM and arrows indicate gB (M and N). The boxed area in (M) is shown in (N). AL cisternae are indicated by white arrows. Gold particles indicating gM (black arrowheads) and gB (black arrows) were localized in a virion and the membrane of the vacuole containing the virion [the boxed area in (M) and white arrowheads in (N)] but not in AL cisternae surrounding the vacuole [white arrows in (N)]. Scale bars: 0.3 μm (A–H), 0.2 μm [inset in (F, I and N)] and 0.5 μm (J–M).
Figure Legend Snippet: Electron micrographs showing the primary envelopment and nuclear egress of HHV-6 capsids and the formation of AL in HHV-6-infected cells A–I) Ultrathin sections of Epon-embedded cells. J–N) Ultrathin cryosections. A–E) Translocation of capsids from the nucleus (n) to the cytoplasm (c). Intranuclear capsids (A), budding of a capsid from the inner nuclear membrane (primary envelopment) (B), a primary enveloped virion in the cisternal space of the nuclear membrane (C), fusion of primary envelope with the outer nuclear membrane (D) and translocation of the capsids into the cytosol (E). F–I) AL were clearly detected near the nuclear membrane of an infected cell (F). The boxed area in (F) is shown in the inset. The outer membrane of the nuclear envelope was closely associated with the membrane of AL [arrowheads in (F) and the inset in (F)]. Membrane association was also seen between AL and the rough endoplasmic reticulum [arrows in (F and G)]. Note that vesicular or tubular membranes that enwrapped or surrounded virions [arrowheads in (H and I): the boxed area in (H)] were distinct from the membranes of AL cisternae [arrows in (H and I)]. Immunolabeling of gM (J and K) or gB (L) (10 nm) and double immunolabeling of gM (15 nm) and gB (10 nm) (M) on ultrathin cryosections. Gold particles indicating gM- and/or gB-labeled virions (v) and small vesicles located in tubulo-vacuoles (asterisks) in the Golgi complex (g) and MVB but not in AL in the cells. Arrowheads indicate gM and arrows indicate gB (M and N). The boxed area in (M) is shown in (N). AL cisternae are indicated by white arrows. Gold particles indicating gM (black arrowheads) and gB (black arrows) were localized in a virion and the membrane of the vacuole containing the virion [the boxed area in (M) and white arrowheads in (N)] but not in AL cisternae surrounding the vacuole [white arrows in (N)]. Scale bars: 0.3 μm (A–H), 0.2 μm [inset in (F, I and N)] and 0.5 μm (J–M).

Techniques Used: Infection, Translocation Assay, Immunolabeling, Labeling

Morphological features of HHV-6-infected HSB-2 cells Ultrathin sections of Epon-embedded cells. Low (A and C) and high [(B) is boxed area in (A) and (D) is boxed area in (C)] power views of HHV6-infected (A and B) and uninfected (C and D) cells. Arrows indicate MVB-like membrane structures around several profiles of the Golgi apparatus (g). Scale bars: 2 μm.
Figure Legend Snippet: Morphological features of HHV-6-infected HSB-2 cells Ultrathin sections of Epon-embedded cells. Low (A and C) and high [(B) is boxed area in (A) and (D) is boxed area in (C)] power views of HHV6-infected (A and B) and uninfected (C and D) cells. Arrows indicate MVB-like membrane structures around several profiles of the Golgi apparatus (g). Scale bars: 2 μm.

Techniques Used: Infection

37) Product Images from "High Resolution Imaging of Temporal and Spatial Changes of Subcellular Ascorbate, Glutathione and H2O2 Distribution during Botrytis cinerea Infection in Arabidopsis"

Article Title: High Resolution Imaging of Temporal and Spatial Changes of Subcellular Ascorbate, Glutathione and H2O2 Distribution during Botrytis cinerea Infection in Arabidopsis

Journal: PLoS ONE

doi: 10.1371/journal.pone.0065811

Changes in the size and fine structure of chloroplasts. Size and fine structures of chloroplasts was evaluated by TEM on longitudinal ultrathin sections within the mesophyll of leaves from Arabidopsis thaliana Col-0 leaves inoculated with Botrytis cinerea and compared to CL. Measurements were performed 0 h ( = CL, represented by the dotted line), 12, 24, 48 and 96 hpi at the IS and the AIS. Shown are means with standard errors. n > 100 chloroplasts per treatment. Significant differences were calculated using the Mann-Whitney U-test; *, **, and ***, respectively, indicate significance at the 0.05, 0.01 and 0.001 levels of confidence. Square = AIS; triangle = IS.
Figure Legend Snippet: Changes in the size and fine structure of chloroplasts. Size and fine structures of chloroplasts was evaluated by TEM on longitudinal ultrathin sections within the mesophyll of leaves from Arabidopsis thaliana Col-0 leaves inoculated with Botrytis cinerea and compared to CL. Measurements were performed 0 h ( = CL, represented by the dotted line), 12, 24, 48 and 96 hpi at the IS and the AIS. Shown are means with standard errors. n > 100 chloroplasts per treatment. Significant differences were calculated using the Mann-Whitney U-test; *, **, and ***, respectively, indicate significance at the 0.05, 0.01 and 0.001 levels of confidence. Square = AIS; triangle = IS.

Techniques Used: Transmission Electron Microscopy, MANN-WHITNEY

38) Product Images from "A barley powdery mildew fungus non-autonomous retrotransposon encodes a peptide that supports penetration success on barley"

Article Title: A barley powdery mildew fungus non-autonomous retrotransposon encodes a peptide that supports penetration success on barley

Journal: Journal of Experimental Botany

doi: 10.1093/jxb/ery174

Immunogold labeling of α-ROPIP1 in Bgh -challenged barley leaves. Transmission electron micrographs of ultrathin sections of Bgh -infected barley epidermal cells 3 dai showing gold particles bound to α-ROPIP1. (A, B) Negative control of infected cells treated with a non-specific antibody. Gold particles were absent in the susceptible barley epidermal cell containing intracellular fungal haustorial protrusions (H) and the extracellular Bgh hypha (Hy). (C, D) Gold particles bound to α-ROPIP1 were observed in hyphae, inside a Bgh appressorium (App), the barley epidermal cell wall (CW), and papilla, but were absent from the extracellular space (ES) and the host cell vacuole (V). (E, F) Gold particles were found in the lumen of finger-like Bgh haustorial protrusions inside barley epidermal cells as well as the host cell cytoplasm, but were almost absent from the host cell vacuole (V), the CW, and the ES. Arrowheads in (D) and (F) point to selected gold particles. Scale bars are 1 µm.
Figure Legend Snippet: Immunogold labeling of α-ROPIP1 in Bgh -challenged barley leaves. Transmission electron micrographs of ultrathin sections of Bgh -infected barley epidermal cells 3 dai showing gold particles bound to α-ROPIP1. (A, B) Negative control of infected cells treated with a non-specific antibody. Gold particles were absent in the susceptible barley epidermal cell containing intracellular fungal haustorial protrusions (H) and the extracellular Bgh hypha (Hy). (C, D) Gold particles bound to α-ROPIP1 were observed in hyphae, inside a Bgh appressorium (App), the barley epidermal cell wall (CW), and papilla, but were absent from the extracellular space (ES) and the host cell vacuole (V). (E, F) Gold particles were found in the lumen of finger-like Bgh haustorial protrusions inside barley epidermal cells as well as the host cell cytoplasm, but were almost absent from the host cell vacuole (V), the CW, and the ES. Arrowheads in (D) and (F) point to selected gold particles. Scale bars are 1 µm.

Techniques Used: Labeling, Transmission Assay, Infection, Negative Control

39) Product Images from "Dynamic compartment specific changes in glutathione and ascorbate levels in Arabidopsis plants exposed to different light intensities"

Article Title: Dynamic compartment specific changes in glutathione and ascorbate levels in Arabidopsis plants exposed to different light intensities

Journal: BMC Plant Biology

doi: 10.1186/1471-2229-13-104

Areas of internal chloroplast structures after the exposure to different light intensities for 14 d. Area (in μm 2 ) and percentage of areas of internal chloroplast structures detected by TEM on a longitudinal ultrathin section within the mesophyll of leaves from the wildtype (Col-0), the pad2-1 and the vtc2-1 mutants after the exposure to different light intensities for 14 d. Data are means with standard errors. Significant differences were calculated within one line of plants between control conditions (exposure to 150 μmol m -2 s -1 ) and the same line exposed to the other light intensities by using the Mann Whitney U-test; *, ** and ***, respectively, indicate significance at the 0.05, 0.01 and 0.001 levels of confidence. n > 100 chloroplasts per treatment.
Figure Legend Snippet: Areas of internal chloroplast structures after the exposure to different light intensities for 14 d. Area (in μm 2 ) and percentage of areas of internal chloroplast structures detected by TEM on a longitudinal ultrathin section within the mesophyll of leaves from the wildtype (Col-0), the pad2-1 and the vtc2-1 mutants after the exposure to different light intensities for 14 d. Data are means with standard errors. Significant differences were calculated within one line of plants between control conditions (exposure to 150 μmol m -2 s -1 ) and the same line exposed to the other light intensities by using the Mann Whitney U-test; *, ** and ***, respectively, indicate significance at the 0.05, 0.01 and 0.001 levels of confidence. n > 100 chloroplasts per treatment.

Techniques Used: Transmission Electron Microscopy, MANN-WHITNEY

40) Product Images from "Type 1 Cannabinoid Receptor-Containing Axons Innervate Hypophysiotropic Thyrotropin-Releasing Hormone-Synthesizing Neurons"

Article Title: Type 1 Cannabinoid Receptor-Containing Axons Innervate Hypophysiotropic Thyrotropin-Releasing Hormone-Synthesizing Neurons

Journal: Endocrinology

doi: 10.1210/en.2008-0330

Electron micrographs of series of ultrathin sections showing an asymmetric-type synaptic contact ( arrows ) between a TRH-containing dendritic spine and a CB1-containing axon terminal. The TRH-IR dendrite (*) and dendritic spine are labeled with
Figure Legend Snippet: Electron micrographs of series of ultrathin sections showing an asymmetric-type synaptic contact ( arrows ) between a TRH-containing dendritic spine and a CB1-containing axon terminal. The TRH-IR dendrite (*) and dendritic spine are labeled with

Techniques Used: Labeling

Related Articles

Staining:

Article Title: Combined genomic and structural analyses of a cultured magnetotactic bacterium reveals its niche adaptation to a dynamic environment
Article Snippet: .. Ultrathin sections were obtained on Leica EM U6 ultramicrotome (Leica Microsystems, Bannockburn, IL, USA), stained with uranyl acetate and lead citrate and imaged with a Morgagni transmission electron microscope (FEI Company, Hillsboro, OR, USA) at 80kV. .. Electron tomography was performed using a JEOL JEM 2100F transmission electron microscope operated at 200 kV using STEM/HAADF mode.

Microscopy:

Article Title: Combined genomic and structural analyses of a cultured magnetotactic bacterium reveals its niche adaptation to a dynamic environment
Article Snippet: .. Ultrathin sections were obtained on Leica EM U6 ultramicrotome (Leica Microsystems, Bannockburn, IL, USA), stained with uranyl acetate and lead citrate and imaged with a Morgagni transmission electron microscope (FEI Company, Hillsboro, OR, USA) at 80kV. .. Electron tomography was performed using a JEOL JEM 2100F transmission electron microscope operated at 200 kV using STEM/HAADF mode.

Transmission Assay:

Article Title: Combined genomic and structural analyses of a cultured magnetotactic bacterium reveals its niche adaptation to a dynamic environment
Article Snippet: .. Ultrathin sections were obtained on Leica EM U6 ultramicrotome (Leica Microsystems, Bannockburn, IL, USA), stained with uranyl acetate and lead citrate and imaged with a Morgagni transmission electron microscope (FEI Company, Hillsboro, OR, USA) at 80kV. .. Electron tomography was performed using a JEOL JEM 2100F transmission electron microscope operated at 200 kV using STEM/HAADF mode.

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    Leica Microsystems ultrathin sections
    Ultrastructure of Mf. australis strain IT-1. Ultrastructure of Mf. australis strain IT-1. a Whole mount TEM image showing a single magnetosome chain, P-rich (P) and sulfur (S) granules; ( b ) <t>Ultrathin</t> section TEM image of high pressure frozen and freeze-substituted cells showing P-rich (P) and sulfur (S) granules, two magnetosomes ( black arrows ), a flagella bundle (F) associated with chemoreceptor array ( white arrows ), and a fibrillar layer at the cell surface ( arrowheads ). Uncharacterized globular structures (G) embedded in an electron-lucent material (asterisks) can be observed
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    Ultrastructure of Mf. australis strain IT-1. Ultrastructure of Mf. australis strain IT-1. a Whole mount TEM image showing a single magnetosome chain, P-rich (P) and sulfur (S) granules; ( b ) Ultrathin section TEM image of high pressure frozen and freeze-substituted cells showing P-rich (P) and sulfur (S) granules, two magnetosomes ( black arrows ), a flagella bundle (F) associated with chemoreceptor array ( white arrows ), and a fibrillar layer at the cell surface ( arrowheads ). Uncharacterized globular structures (G) embedded in an electron-lucent material (asterisks) can be observed

    Journal: BMC Genomics

    Article Title: Combined genomic and structural analyses of a cultured magnetotactic bacterium reveals its niche adaptation to a dynamic environment

    doi: 10.1186/s12864-016-3064-9

    Figure Lengend Snippet: Ultrastructure of Mf. australis strain IT-1. Ultrastructure of Mf. australis strain IT-1. a Whole mount TEM image showing a single magnetosome chain, P-rich (P) and sulfur (S) granules; ( b ) Ultrathin section TEM image of high pressure frozen and freeze-substituted cells showing P-rich (P) and sulfur (S) granules, two magnetosomes ( black arrows ), a flagella bundle (F) associated with chemoreceptor array ( white arrows ), and a fibrillar layer at the cell surface ( arrowheads ). Uncharacterized globular structures (G) embedded in an electron-lucent material (asterisks) can be observed

    Article Snippet: Ultrathin sections were obtained on Leica EM U6 ultramicrotome (Leica Microsystems, Bannockburn, IL, USA), stained with uranyl acetate and lead citrate and imaged with a Morgagni transmission electron microscope (FEI Company, Hillsboro, OR, USA) at 80kV.

    Techniques: Transmission Electron Microscopy

    Transmission electron micrographs of ultrathin sections of E . coli cells from domestic sewage grown in n -hexadecane-free medium (a) and in n -hexadecane-containing medium (b) and of E . coli cells from wheat straw grown in n -hexadecane-free medium (c) and

    Journal: BioMed Research International

    Article Title: Coliform Bacteria for Bioremediation of Waste Hydrocarbons

    doi: 10.1155/2017/1838072

    Figure Lengend Snippet: Transmission electron micrographs of ultrathin sections of E . coli cells from domestic sewage grown in n -hexadecane-free medium (a) and in n -hexadecane-containing medium (b) and of E . coli cells from wheat straw grown in n -hexadecane-free medium (c) and

    Article Snippet: Ultrathin sections [ ] were obtained using an ultramicrotome Leica (Leica Microsystems, Germany).

    Techniques: Transmission Assay

    TEM ultrathin sections of HeLa cells after 12 h incubation with fluorescent magnetopolymersomes (PBD(1200)- b -PEO(600), 10% SPION, 5% DEAC). ( A ) Overview of a typical preparation showing few polymersomes as dark spherical objects and ( B ) peripheral cell region with a rare observation of an internalized magnetopolymersome. The inset depicts a close-up of a multilamellar magnetopolymersome judged from contrast and membrane thickness.

    Journal: Materials

    Article Title: Fluorescent Magnetopolymersomes: A Theranostic Platform to Track Intracellular Delivery

    doi: 10.3390/ma10111303

    Figure Lengend Snippet: TEM ultrathin sections of HeLa cells after 12 h incubation with fluorescent magnetopolymersomes (PBD(1200)- b -PEO(600), 10% SPION, 5% DEAC). ( A ) Overview of a typical preparation showing few polymersomes as dark spherical objects and ( B ) peripheral cell region with a rare observation of an internalized magnetopolymersome. The inset depicts a close-up of a multilamellar magnetopolymersome judged from contrast and membrane thickness.

    Article Snippet: Ultrathin sections were prepared using a Leica Ultracut UC-7 (Leica Microsystems Inc., Buffalo Grove, IL, USA) equipped with a Diatome Ultra 45° diamond knife (DiATOME, Hatfield, PA, USA).

    Techniques: Transmission Electron Microscopy, Incubation

    TEM ultrathin sections of HeLa cells after 12 h incubation with cationic fluorescent magnetopolymersomes. Vesicles (50% PBD- b -PEO-COOH; 2% DEAC) containing 10% w/w SPION were formed at 2 mg/mL via solvent inversion and subsequently coated in 10× b -PEI(800) before being purified by gel permeation chromatography. Overview in ( A ) shows elevated levels of cationically modified magnetopolymersomes taken up (black spots indicated by arrows). The sequence ( B – F ) shows higher magnification images of the apparent progression of hydrolytic degradation of SPION-loaded polymersomes after internalization.

    Journal: Materials

    Article Title: Fluorescent Magnetopolymersomes: A Theranostic Platform to Track Intracellular Delivery

    doi: 10.3390/ma10111303

    Figure Lengend Snippet: TEM ultrathin sections of HeLa cells after 12 h incubation with cationic fluorescent magnetopolymersomes. Vesicles (50% PBD- b -PEO-COOH; 2% DEAC) containing 10% w/w SPION were formed at 2 mg/mL via solvent inversion and subsequently coated in 10× b -PEI(800) before being purified by gel permeation chromatography. Overview in ( A ) shows elevated levels of cationically modified magnetopolymersomes taken up (black spots indicated by arrows). The sequence ( B – F ) shows higher magnification images of the apparent progression of hydrolytic degradation of SPION-loaded polymersomes after internalization.

    Article Snippet: Ultrathin sections were prepared using a Leica Ultracut UC-7 (Leica Microsystems Inc., Buffalo Grove, IL, USA) equipped with a Diatome Ultra 45° diamond knife (DiATOME, Hatfield, PA, USA).

    Techniques: Transmission Electron Microscopy, Incubation, Purification, GPC Assay, Modification, Sequencing

    Immunogold labeling of Lowicryl HM20 sections of HP-frozen, low-temperature embedded immature anthers of Arabidopsis thaliana generated by accelerated FS under agitation. (A) Overview image showing a middle layer cell from an anther surrounded by anti-xyloglucan labeled cell wall (cw). (B) Aggregation of intracellular vesicle-like xyloglucan-positive structures. (C) Overview image showing anti-α-tubulin labeling close to the plasma membrane. (D) Details of immunolabeled cortical microtubules (arrows) aligned almost in parallel to the section plane. Typical for postembedding labeling, the gold-conjugated antibodies are concentrated at regions where the MTs meet the surface of the ultrathin section. Scale bars = 1 µm in A and C; 300 nm in B and D. Abbreviations: n, nucleus; v, vacuole; chl, chloroplast; HP, high-pressure; FS, freeze substitution; MT, microtubule.

    Journal: Journal of Histochemistry and Cytochemistry

    Article Title: Agitation Modules: Flexible Means to Accelerate Automated Freeze Substitution

    doi: 10.1369/0022155418786698

    Figure Lengend Snippet: Immunogold labeling of Lowicryl HM20 sections of HP-frozen, low-temperature embedded immature anthers of Arabidopsis thaliana generated by accelerated FS under agitation. (A) Overview image showing a middle layer cell from an anther surrounded by anti-xyloglucan labeled cell wall (cw). (B) Aggregation of intracellular vesicle-like xyloglucan-positive structures. (C) Overview image showing anti-α-tubulin labeling close to the plasma membrane. (D) Details of immunolabeled cortical microtubules (arrows) aligned almost in parallel to the section plane. Typical for postembedding labeling, the gold-conjugated antibodies are concentrated at regions where the MTs meet the surface of the ultrathin section. Scale bars = 1 µm in A and C; 300 nm in B and D. Abbreviations: n, nucleus; v, vacuole; chl, chloroplast; HP, high-pressure; FS, freeze substitution; MT, microtubule.

    Article Snippet: Ultrathin sections of 40–50 nm were cut with an ultramicrotome UC6 (Leica Microsystems), transferred onto Formvar-coated copper grids, and contrasted with 1% aqueous uranyl acetate for 10 min, followed by Reynolds lead citrate for 10 min.

    Techniques: Labeling, Generated, Immunolabeling