paraformaldehyde pfa Search Results


  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 99
    Millipore paraformaledhyde pfa
    Saccharides and saccharide-modifying enzymes do not affect <t>N-CAM</t> binding to knock-out progenitor cells. Cells were pretreated (1 hr) with heparin (0.01 mg/ml), chondroitin sulfate (0.01 mg/ml), heparinase (0.01 U/ml), or glycosidases (4 U/ml; N-glycosidase endo F and O-glycosidase) and then treated with N-CAM (10 μg/ml) for 2 hr at 37°C. The cells were washed with PBS, fixed with 4% <t>PFA,</t> and processed for immunocytochemistry for N-CAM, as described in Materials and Methods.
    Paraformaledhyde Pfa, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 1830 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/paraformaledhyde pfa/product/Millipore
    Average 99 stars, based on 1830 article reviews
    Price from $9.99 to $1999.99
    paraformaledhyde pfa - by Bioz Stars, 2020-08
    99/100 stars
      Buy from Supplier

    92
    FUJIFILM paraformaldehyde pfa
    Saccharides and saccharide-modifying enzymes do not affect <t>N-CAM</t> binding to knock-out progenitor cells. Cells were pretreated (1 hr) with heparin (0.01 mg/ml), chondroitin sulfate (0.01 mg/ml), heparinase (0.01 U/ml), or glycosidases (4 U/ml; N-glycosidase endo F and O-glycosidase) and then treated with N-CAM (10 μg/ml) for 2 hr at 37°C. The cells were washed with PBS, fixed with 4% <t>PFA,</t> and processed for immunocytochemistry for N-CAM, as described in Materials and Methods.
    Paraformaldehyde Pfa, supplied by FUJIFILM, used in various techniques. Bioz Stars score: 92/100, based on 128 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/paraformaldehyde pfa/product/FUJIFILM
    Average 92 stars, based on 128 article reviews
    Price from $9.99 to $1999.99
    paraformaldehyde pfa - by Bioz Stars, 2020-08
    92/100 stars
      Buy from Supplier

    90
    Junsei Chemical paraformaldehyde pfa
    Saccharides and saccharide-modifying enzymes do not affect <t>N-CAM</t> binding to knock-out progenitor cells. Cells were pretreated (1 hr) with heparin (0.01 mg/ml), chondroitin sulfate (0.01 mg/ml), heparinase (0.01 U/ml), or glycosidases (4 U/ml; N-glycosidase endo F and O-glycosidase) and then treated with N-CAM (10 μg/ml) for 2 hr at 37°C. The cells were washed with PBS, fixed with 4% <t>PFA,</t> and processed for immunocytochemistry for N-CAM, as described in Materials and Methods.
    Paraformaldehyde Pfa, supplied by Junsei Chemical, used in various techniques. Bioz Stars score: 90/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/paraformaldehyde pfa/product/Junsei Chemical
    Average 90 stars, based on 12 article reviews
    Price from $9.99 to $1999.99
    paraformaldehyde pfa - by Bioz Stars, 2020-08
    90/100 stars
      Buy from Supplier

    92
    Carl Roth GmbH paraformaldehyde pfa
    Saccharides and saccharide-modifying enzymes do not affect <t>N-CAM</t> binding to knock-out progenitor cells. Cells were pretreated (1 hr) with heparin (0.01 mg/ml), chondroitin sulfate (0.01 mg/ml), heparinase (0.01 U/ml), or glycosidases (4 U/ml; N-glycosidase endo F and O-glycosidase) and then treated with N-CAM (10 μg/ml) for 2 hr at 37°C. The cells were washed with PBS, fixed with 4% <t>PFA,</t> and processed for immunocytochemistry for N-CAM, as described in Materials and Methods.
    Paraformaldehyde Pfa, supplied by Carl Roth GmbH, used in various techniques. Bioz Stars score: 92/100, based on 96 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/paraformaldehyde pfa/product/Carl Roth GmbH
    Average 92 stars, based on 96 article reviews
    Price from $9.99 to $1999.99
    paraformaldehyde pfa - by Bioz Stars, 2020-08
    92/100 stars
      Buy from Supplier

    92
    Merck KGaA paraformaldehyde pfa
    Protocol for 3D blood vessel visualization in whole mouse brains. (a) Sample preparation: During animal sacrifice mice are transcardially perfused with 40 mL of phosphate-buffered saline <t>(PBS)</t> containing 100 U/mL heparin, followed by 20 mL of 4% paraformaldehyde <t>(PFA)</t> in PBS and 10 mL hydrogel composed of 2% gelatin and 0.1% FITC-conjugated albumin in PBS. (b) Whole brain clearing: In the optimized protocol, the brain specimen is incubated in each clearing solution (tetrahydrofuran (THF) used for dehydration and lipid solvation, dibenzyl ether (DBE) for matching of refractive index) for 12 h at room temperature with constant shaking in dark bottles. To ensure complete dehydration, the 100% THF step was performed twice. (c) Light sheet microscopy: Cleared brains immersed in DBE were imaged in a dorso-ventral direction using either 1.6 × or 6.4 × magnification and corresponding step sizes of 6 µm and 4 µm in the LaVision Biotec ultramicroscope.
    Paraformaldehyde Pfa, supplied by Merck KGaA, used in various techniques. Bioz Stars score: 92/100, based on 362 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/paraformaldehyde pfa/product/Merck KGaA
    Average 92 stars, based on 362 article reviews
    Price from $9.99 to $1999.99
    paraformaldehyde pfa - by Bioz Stars, 2020-08
    92/100 stars
      Buy from Supplier

    92
    Santa Cruz Biotechnology paraformaldehyde pfa
    Protocol for 3D blood vessel visualization in whole mouse brains. (a) Sample preparation: During animal sacrifice mice are transcardially perfused with 40 mL of phosphate-buffered saline <t>(PBS)</t> containing 100 U/mL heparin, followed by 20 mL of 4% paraformaldehyde <t>(PFA)</t> in PBS and 10 mL hydrogel composed of 2% gelatin and 0.1% FITC-conjugated albumin in PBS. (b) Whole brain clearing: In the optimized protocol, the brain specimen is incubated in each clearing solution (tetrahydrofuran (THF) used for dehydration and lipid solvation, dibenzyl ether (DBE) for matching of refractive index) for 12 h at room temperature with constant shaking in dark bottles. To ensure complete dehydration, the 100% THF step was performed twice. (c) Light sheet microscopy: Cleared brains immersed in DBE were imaged in a dorso-ventral direction using either 1.6 × or 6.4 × magnification and corresponding step sizes of 6 µm and 4 µm in the LaVision Biotec ultramicroscope.
    Paraformaldehyde Pfa, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 92/100, based on 428 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/paraformaldehyde pfa/product/Santa Cruz Biotechnology
    Average 92 stars, based on 428 article reviews
    Price from $9.99 to $1999.99
    paraformaldehyde pfa - by Bioz Stars, 2020-08
    92/100 stars
      Buy from Supplier

    93
    Merck & Co paraformaldehyde pfa
    Protocol for 3D blood vessel visualization in whole mouse brains. (a) Sample preparation: During animal sacrifice mice are transcardially perfused with 40 mL of phosphate-buffered saline <t>(PBS)</t> containing 100 U/mL heparin, followed by 20 mL of 4% paraformaldehyde <t>(PFA)</t> in PBS and 10 mL hydrogel composed of 2% gelatin and 0.1% FITC-conjugated albumin in PBS. (b) Whole brain clearing: In the optimized protocol, the brain specimen is incubated in each clearing solution (tetrahydrofuran (THF) used for dehydration and lipid solvation, dibenzyl ether (DBE) for matching of refractive index) for 12 h at room temperature with constant shaking in dark bottles. To ensure complete dehydration, the 100% THF step was performed twice. (c) Light sheet microscopy: Cleared brains immersed in DBE were imaged in a dorso-ventral direction using either 1.6 × or 6.4 × magnification and corresponding step sizes of 6 µm and 4 µm in the LaVision Biotec ultramicroscope.
    Paraformaldehyde Pfa, supplied by Merck & Co, used in various techniques. Bioz Stars score: 93/100, based on 159 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/paraformaldehyde pfa/product/Merck & Co
    Average 93 stars, based on 159 article reviews
    Price from $9.99 to $1999.99
    paraformaldehyde pfa - by Bioz Stars, 2020-08
    93/100 stars
      Buy from Supplier

    94
    Thermo Fisher paraformaldehyde pfa
    Strategies for achieving single-molecule visualization and cellular structural preservation in S. pombe cells. (a) The expression patterns of <t>Ost4-GFP,</t> Ost4-GFP-MTn, Nup124-GFP, Nup124-GFP-MTn, Sad1-GFP, and Sad1-GFP-MTn in S. pombe observed by live-cell fluorescence imaging (scale bar, 2 μm). The fluorescence images of GFP showed that the fusion proteins were correctly localized to expected subcellular locations: membranes of cortical ER and NE for Ost4, NPs for Nup124, the SPB for Sad1. (b) A cartoon shows the NE and cortical ER, NPs, and the SPB in a S. pombe cell. The regions of interest are labeled with numbers (I-VI) as references in other figures. (c) Procedures for the preparation of S. pombe cells for synthesizing AuNPs, three schemes labeled as 1 (“Cold MeOH”), 2 (“Oxidization + Chemical Fixation + Permeabilization”), 3 (“HPF/FSF + Oxidization + Chemical Fixation + Rehydration”) respectively, the Scheme 2 consists of four different sub-schemes (2a-2d), see the Supplementary Table 3 for details. (d) Synthesis of AuNPs in S. pombe cells treated by the “Cold MeOH” protocol with the optimized ANSM-based AuNPs synthesis protocol obtained with E. coli cells ( Fig.3b ) , the upper row shows the S. pombe cell suspension after AuNPs synthesis and the lower row shows the corresponding centrifuged cell pellets, apparently the colors of those MTn tags expressed cells are much darker than the corresponding control cells. (e) A typical EM image showing the unprecedented high labelling densities of AuNPs distributed on the cortical ER (II) and ER stacks (III) in a cell expressing Ost4-GFP-MTn prepared with the Scheme 1. (f) EM image showing better membranes preservation of an Ost4-GFP-MTn expressed cell treated with Scheme 2c than that in (e), the AuNPs could be clearly localized to outer membranes of the NE although the labeling densities of AuNPs reduced after the <t>PFA</t> fixation. (g) EM image showing the linear distributions of AuNPs along the membranes of cortical ER and ER stacks in a cell expressing Ost4-GFP-MTn treated with the Scheme 2a (with cold MeOH permeabilization), the structural preservation is better than that in (e) but with reduced labelling densities of AuNPs after the GA fixation. (h-i) EM images showing the AuNPs distributed along the membranes of NE (I) and ERs (II, IV) in cells expressing Ost4-GFP-MTn treated with Scheme 2b, lack of AuNPs in the nuclear pores (NPs), only a few background AuNPs scattered in cytosols, nuclei and mitochondria (M). (j) high densities of AuNPs formed on ER stacks in cells expressing Ost4-GFP-MTα treated with Scheme 2b (with 0.1% Triton X-100 2 min permeabilization). (k) A lot of AuNPs gathered in the SPB (among a dark density region), the specimen was prepared with Scheme 2a. (l) AuNPs synthesized in a cell expressing Nup124-GFP-MTn with Scheme 3a, grouped AuNPs formed on the NP (dark regions) with a regular interval along the NE. (m) Statistical analysis (one-way ANOVA followed by Brown-Forsythe test) of the AuNPs densities in ER (including NE), cytosol (Cyto), Nucleus (Nu) and mitochondrion (Mito) based on 20 EM images of 90 nm thick sections of S. pombe cells expressing Ost4-GFP-MTn (see Supplementary Chart 1 ).
    Paraformaldehyde Pfa, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 94/100, based on 3743 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/paraformaldehyde pfa/product/Thermo Fisher
    Average 94 stars, based on 3743 article reviews
    Price from $9.99 to $1999.99
    paraformaldehyde pfa - by Bioz Stars, 2020-08
    94/100 stars
      Buy from Supplier

    92
    Polysciences inc paraformaldehyde pfa
    Strategies for achieving single-molecule visualization and cellular structural preservation in S. pombe cells. (a) The expression patterns of <t>Ost4-GFP,</t> Ost4-GFP-MTn, Nup124-GFP, Nup124-GFP-MTn, Sad1-GFP, and Sad1-GFP-MTn in S. pombe observed by live-cell fluorescence imaging (scale bar, 2 μm). The fluorescence images of GFP showed that the fusion proteins were correctly localized to expected subcellular locations: membranes of cortical ER and NE for Ost4, NPs for Nup124, the SPB for Sad1. (b) A cartoon shows the NE and cortical ER, NPs, and the SPB in a S. pombe cell. The regions of interest are labeled with numbers (I-VI) as references in other figures. (c) Procedures for the preparation of S. pombe cells for synthesizing AuNPs, three schemes labeled as 1 (“Cold MeOH”), 2 (“Oxidization + Chemical Fixation + Permeabilization”), 3 (“HPF/FSF + Oxidization + Chemical Fixation + Rehydration”) respectively, the Scheme 2 consists of four different sub-schemes (2a-2d), see the Supplementary Table 3 for details. (d) Synthesis of AuNPs in S. pombe cells treated by the “Cold MeOH” protocol with the optimized ANSM-based AuNPs synthesis protocol obtained with E. coli cells ( Fig.3b ) , the upper row shows the S. pombe cell suspension after AuNPs synthesis and the lower row shows the corresponding centrifuged cell pellets, apparently the colors of those MTn tags expressed cells are much darker than the corresponding control cells. (e) A typical EM image showing the unprecedented high labelling densities of AuNPs distributed on the cortical ER (II) and ER stacks (III) in a cell expressing Ost4-GFP-MTn prepared with the Scheme 1. (f) EM image showing better membranes preservation of an Ost4-GFP-MTn expressed cell treated with Scheme 2c than that in (e), the AuNPs could be clearly localized to outer membranes of the NE although the labeling densities of AuNPs reduced after the <t>PFA</t> fixation. (g) EM image showing the linear distributions of AuNPs along the membranes of cortical ER and ER stacks in a cell expressing Ost4-GFP-MTn treated with the Scheme 2a (with cold MeOH permeabilization), the structural preservation is better than that in (e) but with reduced labelling densities of AuNPs after the GA fixation. (h-i) EM images showing the AuNPs distributed along the membranes of NE (I) and ERs (II, IV) in cells expressing Ost4-GFP-MTn treated with Scheme 2b, lack of AuNPs in the nuclear pores (NPs), only a few background AuNPs scattered in cytosols, nuclei and mitochondria (M). (j) high densities of AuNPs formed on ER stacks in cells expressing Ost4-GFP-MTα treated with Scheme 2b (with 0.1% Triton X-100 2 min permeabilization). (k) A lot of AuNPs gathered in the SPB (among a dark density region), the specimen was prepared with Scheme 2a. (l) AuNPs synthesized in a cell expressing Nup124-GFP-MTn with Scheme 3a, grouped AuNPs formed on the NP (dark regions) with a regular interval along the NE. (m) Statistical analysis (one-way ANOVA followed by Brown-Forsythe test) of the AuNPs densities in ER (including NE), cytosol (Cyto), Nucleus (Nu) and mitochondrion (Mito) based on 20 EM images of 90 nm thick sections of S. pombe cells expressing Ost4-GFP-MTn (see Supplementary Chart 1 ).
    Paraformaldehyde Pfa, supplied by Polysciences inc, used in various techniques. Bioz Stars score: 92/100, based on 99 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/paraformaldehyde pfa/product/Polysciences inc
    Average 92 stars, based on 99 article reviews
    Price from $9.99 to $1999.99
    paraformaldehyde pfa - by Bioz Stars, 2020-08
    92/100 stars
      Buy from Supplier

    93
    Fisher Scientific paraformaldehyde pfa
    Strategies for achieving single-molecule visualization and cellular structural preservation in S. pombe cells. (a) The expression patterns of <t>Ost4-GFP,</t> Ost4-GFP-MTn, Nup124-GFP, Nup124-GFP-MTn, Sad1-GFP, and Sad1-GFP-MTn in S. pombe observed by live-cell fluorescence imaging (scale bar, 2 μm). The fluorescence images of GFP showed that the fusion proteins were correctly localized to expected subcellular locations: membranes of cortical ER and NE for Ost4, NPs for Nup124, the SPB for Sad1. (b) A cartoon shows the NE and cortical ER, NPs, and the SPB in a S. pombe cell. The regions of interest are labeled with numbers (I-VI) as references in other figures. (c) Procedures for the preparation of S. pombe cells for synthesizing AuNPs, three schemes labeled as 1 (“Cold MeOH”), 2 (“Oxidization + Chemical Fixation + Permeabilization”), 3 (“HPF/FSF + Oxidization + Chemical Fixation + Rehydration”) respectively, the Scheme 2 consists of four different sub-schemes (2a-2d), see the Supplementary Table 3 for details. (d) Synthesis of AuNPs in S. pombe cells treated by the “Cold MeOH” protocol with the optimized ANSM-based AuNPs synthesis protocol obtained with E. coli cells ( Fig.3b ) , the upper row shows the S. pombe cell suspension after AuNPs synthesis and the lower row shows the corresponding centrifuged cell pellets, apparently the colors of those MTn tags expressed cells are much darker than the corresponding control cells. (e) A typical EM image showing the unprecedented high labelling densities of AuNPs distributed on the cortical ER (II) and ER stacks (III) in a cell expressing Ost4-GFP-MTn prepared with the Scheme 1. (f) EM image showing better membranes preservation of an Ost4-GFP-MTn expressed cell treated with Scheme 2c than that in (e), the AuNPs could be clearly localized to outer membranes of the NE although the labeling densities of AuNPs reduced after the <t>PFA</t> fixation. (g) EM image showing the linear distributions of AuNPs along the membranes of cortical ER and ER stacks in a cell expressing Ost4-GFP-MTn treated with the Scheme 2a (with cold MeOH permeabilization), the structural preservation is better than that in (e) but with reduced labelling densities of AuNPs after the GA fixation. (h-i) EM images showing the AuNPs distributed along the membranes of NE (I) and ERs (II, IV) in cells expressing Ost4-GFP-MTn treated with Scheme 2b, lack of AuNPs in the nuclear pores (NPs), only a few background AuNPs scattered in cytosols, nuclei and mitochondria (M). (j) high densities of AuNPs formed on ER stacks in cells expressing Ost4-GFP-MTα treated with Scheme 2b (with 0.1% Triton X-100 2 min permeabilization). (k) A lot of AuNPs gathered in the SPB (among a dark density region), the specimen was prepared with Scheme 2a. (l) AuNPs synthesized in a cell expressing Nup124-GFP-MTn with Scheme 3a, grouped AuNPs formed on the NP (dark regions) with a regular interval along the NE. (m) Statistical analysis (one-way ANOVA followed by Brown-Forsythe test) of the AuNPs densities in ER (including NE), cytosol (Cyto), Nucleus (Nu) and mitochondrion (Mito) based on 20 EM images of 90 nm thick sections of S. pombe cells expressing Ost4-GFP-MTn (see Supplementary Chart 1 ).
    Paraformaldehyde Pfa, supplied by Fisher Scientific, used in various techniques. Bioz Stars score: 93/100, based on 279 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/paraformaldehyde pfa/product/Fisher Scientific
    Average 93 stars, based on 279 article reviews
    Price from $9.99 to $1999.99
    paraformaldehyde pfa - by Bioz Stars, 2020-08
    93/100 stars
      Buy from Supplier

    92
    Ricca Chemical Company paraformaldehyde pfa
    Strategies for achieving single-molecule visualization and cellular structural preservation in S. pombe cells. (a) The expression patterns of <t>Ost4-GFP,</t> Ost4-GFP-MTn, Nup124-GFP, Nup124-GFP-MTn, Sad1-GFP, and Sad1-GFP-MTn in S. pombe observed by live-cell fluorescence imaging (scale bar, 2 μm). The fluorescence images of GFP showed that the fusion proteins were correctly localized to expected subcellular locations: membranes of cortical ER and NE for Ost4, NPs for Nup124, the SPB for Sad1. (b) A cartoon shows the NE and cortical ER, NPs, and the SPB in a S. pombe cell. The regions of interest are labeled with numbers (I-VI) as references in other figures. (c) Procedures for the preparation of S. pombe cells for synthesizing AuNPs, three schemes labeled as 1 (“Cold MeOH”), 2 (“Oxidization + Chemical Fixation + Permeabilization”), 3 (“HPF/FSF + Oxidization + Chemical Fixation + Rehydration”) respectively, the Scheme 2 consists of four different sub-schemes (2a-2d), see the Supplementary Table 3 for details. (d) Synthesis of AuNPs in S. pombe cells treated by the “Cold MeOH” protocol with the optimized ANSM-based AuNPs synthesis protocol obtained with E. coli cells ( Fig.3b ) , the upper row shows the S. pombe cell suspension after AuNPs synthesis and the lower row shows the corresponding centrifuged cell pellets, apparently the colors of those MTn tags expressed cells are much darker than the corresponding control cells. (e) A typical EM image showing the unprecedented high labelling densities of AuNPs distributed on the cortical ER (II) and ER stacks (III) in a cell expressing Ost4-GFP-MTn prepared with the Scheme 1. (f) EM image showing better membranes preservation of an Ost4-GFP-MTn expressed cell treated with Scheme 2c than that in (e), the AuNPs could be clearly localized to outer membranes of the NE although the labeling densities of AuNPs reduced after the <t>PFA</t> fixation. (g) EM image showing the linear distributions of AuNPs along the membranes of cortical ER and ER stacks in a cell expressing Ost4-GFP-MTn treated with the Scheme 2a (with cold MeOH permeabilization), the structural preservation is better than that in (e) but with reduced labelling densities of AuNPs after the GA fixation. (h-i) EM images showing the AuNPs distributed along the membranes of NE (I) and ERs (II, IV) in cells expressing Ost4-GFP-MTn treated with Scheme 2b, lack of AuNPs in the nuclear pores (NPs), only a few background AuNPs scattered in cytosols, nuclei and mitochondria (M). (j) high densities of AuNPs formed on ER stacks in cells expressing Ost4-GFP-MTα treated with Scheme 2b (with 0.1% Triton X-100 2 min permeabilization). (k) A lot of AuNPs gathered in the SPB (among a dark density region), the specimen was prepared with Scheme 2a. (l) AuNPs synthesized in a cell expressing Nup124-GFP-MTn with Scheme 3a, grouped AuNPs formed on the NP (dark regions) with a regular interval along the NE. (m) Statistical analysis (one-way ANOVA followed by Brown-Forsythe test) of the AuNPs densities in ER (including NE), cytosol (Cyto), Nucleus (Nu) and mitochondrion (Mito) based on 20 EM images of 90 nm thick sections of S. pombe cells expressing Ost4-GFP-MTn (see Supplementary Chart 1 ).
    Paraformaldehyde Pfa, supplied by Ricca Chemical Company, used in various techniques. Bioz Stars score: 92/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/paraformaldehyde pfa/product/Ricca Chemical Company
    Average 92 stars, based on 3 article reviews
    Price from $9.99 to $1999.99
    paraformaldehyde pfa - by Bioz Stars, 2020-08
    92/100 stars
      Buy from Supplier

    92
    TAAB Laboratories paraformaldehyde pfa
    Strategies for achieving single-molecule visualization and cellular structural preservation in S. pombe cells. (a) The expression patterns of <t>Ost4-GFP,</t> Ost4-GFP-MTn, Nup124-GFP, Nup124-GFP-MTn, Sad1-GFP, and Sad1-GFP-MTn in S. pombe observed by live-cell fluorescence imaging (scale bar, 2 μm). The fluorescence images of GFP showed that the fusion proteins were correctly localized to expected subcellular locations: membranes of cortical ER and NE for Ost4, NPs for Nup124, the SPB for Sad1. (b) A cartoon shows the NE and cortical ER, NPs, and the SPB in a S. pombe cell. The regions of interest are labeled with numbers (I-VI) as references in other figures. (c) Procedures for the preparation of S. pombe cells for synthesizing AuNPs, three schemes labeled as 1 (“Cold MeOH”), 2 (“Oxidization + Chemical Fixation + Permeabilization”), 3 (“HPF/FSF + Oxidization + Chemical Fixation + Rehydration”) respectively, the Scheme 2 consists of four different sub-schemes (2a-2d), see the Supplementary Table 3 for details. (d) Synthesis of AuNPs in S. pombe cells treated by the “Cold MeOH” protocol with the optimized ANSM-based AuNPs synthesis protocol obtained with E. coli cells ( Fig.3b ) , the upper row shows the S. pombe cell suspension after AuNPs synthesis and the lower row shows the corresponding centrifuged cell pellets, apparently the colors of those MTn tags expressed cells are much darker than the corresponding control cells. (e) A typical EM image showing the unprecedented high labelling densities of AuNPs distributed on the cortical ER (II) and ER stacks (III) in a cell expressing Ost4-GFP-MTn prepared with the Scheme 1. (f) EM image showing better membranes preservation of an Ost4-GFP-MTn expressed cell treated with Scheme 2c than that in (e), the AuNPs could be clearly localized to outer membranes of the NE although the labeling densities of AuNPs reduced after the <t>PFA</t> fixation. (g) EM image showing the linear distributions of AuNPs along the membranes of cortical ER and ER stacks in a cell expressing Ost4-GFP-MTn treated with the Scheme 2a (with cold MeOH permeabilization), the structural preservation is better than that in (e) but with reduced labelling densities of AuNPs after the GA fixation. (h-i) EM images showing the AuNPs distributed along the membranes of NE (I) and ERs (II, IV) in cells expressing Ost4-GFP-MTn treated with Scheme 2b, lack of AuNPs in the nuclear pores (NPs), only a few background AuNPs scattered in cytosols, nuclei and mitochondria (M). (j) high densities of AuNPs formed on ER stacks in cells expressing Ost4-GFP-MTα treated with Scheme 2b (with 0.1% Triton X-100 2 min permeabilization). (k) A lot of AuNPs gathered in the SPB (among a dark density region), the specimen was prepared with Scheme 2a. (l) AuNPs synthesized in a cell expressing Nup124-GFP-MTn with Scheme 3a, grouped AuNPs formed on the NP (dark regions) with a regular interval along the NE. (m) Statistical analysis (one-way ANOVA followed by Brown-Forsythe test) of the AuNPs densities in ER (including NE), cytosol (Cyto), Nucleus (Nu) and mitochondrion (Mito) based on 20 EM images of 90 nm thick sections of S. pombe cells expressing Ost4-GFP-MTn (see Supplementary Chart 1 ).
    Paraformaldehyde Pfa, supplied by TAAB Laboratories, used in various techniques. Bioz Stars score: 92/100, based on 13 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/paraformaldehyde pfa/product/TAAB Laboratories
    Average 92 stars, based on 13 article reviews
    Price from $9.99 to $1999.99
    paraformaldehyde pfa - by Bioz Stars, 2020-08
    92/100 stars
      Buy from Supplier

    92
    Beyotime paraformaldehyde pfa
    Strategies for achieving single-molecule visualization and cellular structural preservation in S. pombe cells. (a) The expression patterns of <t>Ost4-GFP,</t> Ost4-GFP-MTn, Nup124-GFP, Nup124-GFP-MTn, Sad1-GFP, and Sad1-GFP-MTn in S. pombe observed by live-cell fluorescence imaging (scale bar, 2 μm). The fluorescence images of GFP showed that the fusion proteins were correctly localized to expected subcellular locations: membranes of cortical ER and NE for Ost4, NPs for Nup124, the SPB for Sad1. (b) A cartoon shows the NE and cortical ER, NPs, and the SPB in a S. pombe cell. The regions of interest are labeled with numbers (I-VI) as references in other figures. (c) Procedures for the preparation of S. pombe cells for synthesizing AuNPs, three schemes labeled as 1 (“Cold MeOH”), 2 (“Oxidization + Chemical Fixation + Permeabilization”), 3 (“HPF/FSF + Oxidization + Chemical Fixation + Rehydration”) respectively, the Scheme 2 consists of four different sub-schemes (2a-2d), see the Supplementary Table 3 for details. (d) Synthesis of AuNPs in S. pombe cells treated by the “Cold MeOH” protocol with the optimized ANSM-based AuNPs synthesis protocol obtained with E. coli cells ( Fig.3b ) , the upper row shows the S. pombe cell suspension after AuNPs synthesis and the lower row shows the corresponding centrifuged cell pellets, apparently the colors of those MTn tags expressed cells are much darker than the corresponding control cells. (e) A typical EM image showing the unprecedented high labelling densities of AuNPs distributed on the cortical ER (II) and ER stacks (III) in a cell expressing Ost4-GFP-MTn prepared with the Scheme 1. (f) EM image showing better membranes preservation of an Ost4-GFP-MTn expressed cell treated with Scheme 2c than that in (e), the AuNPs could be clearly localized to outer membranes of the NE although the labeling densities of AuNPs reduced after the <t>PFA</t> fixation. (g) EM image showing the linear distributions of AuNPs along the membranes of cortical ER and ER stacks in a cell expressing Ost4-GFP-MTn treated with the Scheme 2a (with cold MeOH permeabilization), the structural preservation is better than that in (e) but with reduced labelling densities of AuNPs after the GA fixation. (h-i) EM images showing the AuNPs distributed along the membranes of NE (I) and ERs (II, IV) in cells expressing Ost4-GFP-MTn treated with Scheme 2b, lack of AuNPs in the nuclear pores (NPs), only a few background AuNPs scattered in cytosols, nuclei and mitochondria (M). (j) high densities of AuNPs formed on ER stacks in cells expressing Ost4-GFP-MTα treated with Scheme 2b (with 0.1% Triton X-100 2 min permeabilization). (k) A lot of AuNPs gathered in the SPB (among a dark density region), the specimen was prepared with Scheme 2a. (l) AuNPs synthesized in a cell expressing Nup124-GFP-MTn with Scheme 3a, grouped AuNPs formed on the NP (dark regions) with a regular interval along the NE. (m) Statistical analysis (one-way ANOVA followed by Brown-Forsythe test) of the AuNPs densities in ER (including NE), cytosol (Cyto), Nucleus (Nu) and mitochondrion (Mito) based on 20 EM images of 90 nm thick sections of S. pombe cells expressing Ost4-GFP-MTn (see Supplementary Chart 1 ).
    Paraformaldehyde Pfa, supplied by Beyotime, used in various techniques. Bioz Stars score: 92/100, based on 119 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/paraformaldehyde pfa/product/Beyotime
    Average 92 stars, based on 119 article reviews
    Price from $9.99 to $1999.99
    paraformaldehyde pfa - by Bioz Stars, 2020-08
    92/100 stars
      Buy from Supplier

    93
    Nacalai paraformaldehyde pfa
    Strategies for achieving single-molecule visualization and cellular structural preservation in S. pombe cells. (a) The expression patterns of <t>Ost4-GFP,</t> Ost4-GFP-MTn, Nup124-GFP, Nup124-GFP-MTn, Sad1-GFP, and Sad1-GFP-MTn in S. pombe observed by live-cell fluorescence imaging (scale bar, 2 μm). The fluorescence images of GFP showed that the fusion proteins were correctly localized to expected subcellular locations: membranes of cortical ER and NE for Ost4, NPs for Nup124, the SPB for Sad1. (b) A cartoon shows the NE and cortical ER, NPs, and the SPB in a S. pombe cell. The regions of interest are labeled with numbers (I-VI) as references in other figures. (c) Procedures for the preparation of S. pombe cells for synthesizing AuNPs, three schemes labeled as 1 (“Cold MeOH”), 2 (“Oxidization + Chemical Fixation + Permeabilization”), 3 (“HPF/FSF + Oxidization + Chemical Fixation + Rehydration”) respectively, the Scheme 2 consists of four different sub-schemes (2a-2d), see the Supplementary Table 3 for details. (d) Synthesis of AuNPs in S. pombe cells treated by the “Cold MeOH” protocol with the optimized ANSM-based AuNPs synthesis protocol obtained with E. coli cells ( Fig.3b ) , the upper row shows the S. pombe cell suspension after AuNPs synthesis and the lower row shows the corresponding centrifuged cell pellets, apparently the colors of those MTn tags expressed cells are much darker than the corresponding control cells. (e) A typical EM image showing the unprecedented high labelling densities of AuNPs distributed on the cortical ER (II) and ER stacks (III) in a cell expressing Ost4-GFP-MTn prepared with the Scheme 1. (f) EM image showing better membranes preservation of an Ost4-GFP-MTn expressed cell treated with Scheme 2c than that in (e), the AuNPs could be clearly localized to outer membranes of the NE although the labeling densities of AuNPs reduced after the <t>PFA</t> fixation. (g) EM image showing the linear distributions of AuNPs along the membranes of cortical ER and ER stacks in a cell expressing Ost4-GFP-MTn treated with the Scheme 2a (with cold MeOH permeabilization), the structural preservation is better than that in (e) but with reduced labelling densities of AuNPs after the GA fixation. (h-i) EM images showing the AuNPs distributed along the membranes of NE (I) and ERs (II, IV) in cells expressing Ost4-GFP-MTn treated with Scheme 2b, lack of AuNPs in the nuclear pores (NPs), only a few background AuNPs scattered in cytosols, nuclei and mitochondria (M). (j) high densities of AuNPs formed on ER stacks in cells expressing Ost4-GFP-MTα treated with Scheme 2b (with 0.1% Triton X-100 2 min permeabilization). (k) A lot of AuNPs gathered in the SPB (among a dark density region), the specimen was prepared with Scheme 2a. (l) AuNPs synthesized in a cell expressing Nup124-GFP-MTn with Scheme 3a, grouped AuNPs formed on the NP (dark regions) with a regular interval along the NE. (m) Statistical analysis (one-way ANOVA followed by Brown-Forsythe test) of the AuNPs densities in ER (including NE), cytosol (Cyto), Nucleus (Nu) and mitochondrion (Mito) based on 20 EM images of 90 nm thick sections of S. pombe cells expressing Ost4-GFP-MTn (see Supplementary Chart 1 ).
    Paraformaldehyde Pfa, supplied by Nacalai, used in various techniques. Bioz Stars score: 93/100, based on 129 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/paraformaldehyde pfa/product/Nacalai
    Average 93 stars, based on 129 article reviews
    Price from $9.99 to $1999.99
    paraformaldehyde pfa - by Bioz Stars, 2020-08
    93/100 stars
      Buy from Supplier

    93
    Avantor paraformaldehyde pfa
    Strategies for achieving single-molecule visualization and cellular structural preservation in S. pombe cells. (a) The expression patterns of <t>Ost4-GFP,</t> Ost4-GFP-MTn, Nup124-GFP, Nup124-GFP-MTn, Sad1-GFP, and Sad1-GFP-MTn in S. pombe observed by live-cell fluorescence imaging (scale bar, 2 μm). The fluorescence images of GFP showed that the fusion proteins were correctly localized to expected subcellular locations: membranes of cortical ER and NE for Ost4, NPs for Nup124, the SPB for Sad1. (b) A cartoon shows the NE and cortical ER, NPs, and the SPB in a S. pombe cell. The regions of interest are labeled with numbers (I-VI) as references in other figures. (c) Procedures for the preparation of S. pombe cells for synthesizing AuNPs, three schemes labeled as 1 (“Cold MeOH”), 2 (“Oxidization + Chemical Fixation + Permeabilization”), 3 (“HPF/FSF + Oxidization + Chemical Fixation + Rehydration”) respectively, the Scheme 2 consists of four different sub-schemes (2a-2d), see the Supplementary Table 3 for details. (d) Synthesis of AuNPs in S. pombe cells treated by the “Cold MeOH” protocol with the optimized ANSM-based AuNPs synthesis protocol obtained with E. coli cells ( Fig.3b ) , the upper row shows the S. pombe cell suspension after AuNPs synthesis and the lower row shows the corresponding centrifuged cell pellets, apparently the colors of those MTn tags expressed cells are much darker than the corresponding control cells. (e) A typical EM image showing the unprecedented high labelling densities of AuNPs distributed on the cortical ER (II) and ER stacks (III) in a cell expressing Ost4-GFP-MTn prepared with the Scheme 1. (f) EM image showing better membranes preservation of an Ost4-GFP-MTn expressed cell treated with Scheme 2c than that in (e), the AuNPs could be clearly localized to outer membranes of the NE although the labeling densities of AuNPs reduced after the <t>PFA</t> fixation. (g) EM image showing the linear distributions of AuNPs along the membranes of cortical ER and ER stacks in a cell expressing Ost4-GFP-MTn treated with the Scheme 2a (with cold MeOH permeabilization), the structural preservation is better than that in (e) but with reduced labelling densities of AuNPs after the GA fixation. (h-i) EM images showing the AuNPs distributed along the membranes of NE (I) and ERs (II, IV) in cells expressing Ost4-GFP-MTn treated with Scheme 2b, lack of AuNPs in the nuclear pores (NPs), only a few background AuNPs scattered in cytosols, nuclei and mitochondria (M). (j) high densities of AuNPs formed on ER stacks in cells expressing Ost4-GFP-MTα treated with Scheme 2b (with 0.1% Triton X-100 2 min permeabilization). (k) A lot of AuNPs gathered in the SPB (among a dark density region), the specimen was prepared with Scheme 2a. (l) AuNPs synthesized in a cell expressing Nup124-GFP-MTn with Scheme 3a, grouped AuNPs formed on the NP (dark regions) with a regular interval along the NE. (m) Statistical analysis (one-way ANOVA followed by Brown-Forsythe test) of the AuNPs densities in ER (including NE), cytosol (Cyto), Nucleus (Nu) and mitochondrion (Mito) based on 20 EM images of 90 nm thick sections of S. pombe cells expressing Ost4-GFP-MTn (see Supplementary Chart 1 ).
    Paraformaldehyde Pfa, supplied by Avantor, used in various techniques. Bioz Stars score: 93/100, based on 254 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/paraformaldehyde pfa/product/Avantor
    Average 93 stars, based on 254 article reviews
    Price from $9.99 to $1999.99
    paraformaldehyde pfa - by Bioz Stars, 2020-08
    93/100 stars
      Buy from Supplier

    92
    EM Science Inc paraformaldehyde pfa
    Strategies for achieving single-molecule visualization and cellular structural preservation in S. pombe cells. (a) The expression patterns of <t>Ost4-GFP,</t> Ost4-GFP-MTn, Nup124-GFP, Nup124-GFP-MTn, Sad1-GFP, and Sad1-GFP-MTn in S. pombe observed by live-cell fluorescence imaging (scale bar, 2 μm). The fluorescence images of GFP showed that the fusion proteins were correctly localized to expected subcellular locations: membranes of cortical ER and NE for Ost4, NPs for Nup124, the SPB for Sad1. (b) A cartoon shows the NE and cortical ER, NPs, and the SPB in a S. pombe cell. The regions of interest are labeled with numbers (I-VI) as references in other figures. (c) Procedures for the preparation of S. pombe cells for synthesizing AuNPs, three schemes labeled as 1 (“Cold MeOH”), 2 (“Oxidization + Chemical Fixation + Permeabilization”), 3 (“HPF/FSF + Oxidization + Chemical Fixation + Rehydration”) respectively, the Scheme 2 consists of four different sub-schemes (2a-2d), see the Supplementary Table 3 for details. (d) Synthesis of AuNPs in S. pombe cells treated by the “Cold MeOH” protocol with the optimized ANSM-based AuNPs synthesis protocol obtained with E. coli cells ( Fig.3b ) , the upper row shows the S. pombe cell suspension after AuNPs synthesis and the lower row shows the corresponding centrifuged cell pellets, apparently the colors of those MTn tags expressed cells are much darker than the corresponding control cells. (e) A typical EM image showing the unprecedented high labelling densities of AuNPs distributed on the cortical ER (II) and ER stacks (III) in a cell expressing Ost4-GFP-MTn prepared with the Scheme 1. (f) EM image showing better membranes preservation of an Ost4-GFP-MTn expressed cell treated with Scheme 2c than that in (e), the AuNPs could be clearly localized to outer membranes of the NE although the labeling densities of AuNPs reduced after the <t>PFA</t> fixation. (g) EM image showing the linear distributions of AuNPs along the membranes of cortical ER and ER stacks in a cell expressing Ost4-GFP-MTn treated with the Scheme 2a (with cold MeOH permeabilization), the structural preservation is better than that in (e) but with reduced labelling densities of AuNPs after the GA fixation. (h-i) EM images showing the AuNPs distributed along the membranes of NE (I) and ERs (II, IV) in cells expressing Ost4-GFP-MTn treated with Scheme 2b, lack of AuNPs in the nuclear pores (NPs), only a few background AuNPs scattered in cytosols, nuclei and mitochondria (M). (j) high densities of AuNPs formed on ER stacks in cells expressing Ost4-GFP-MTα treated with Scheme 2b (with 0.1% Triton X-100 2 min permeabilization). (k) A lot of AuNPs gathered in the SPB (among a dark density region), the specimen was prepared with Scheme 2a. (l) AuNPs synthesized in a cell expressing Nup124-GFP-MTn with Scheme 3a, grouped AuNPs formed on the NP (dark regions) with a regular interval along the NE. (m) Statistical analysis (one-way ANOVA followed by Brown-Forsythe test) of the AuNPs densities in ER (including NE), cytosol (Cyto), Nucleus (Nu) and mitochondrion (Mito) based on 20 EM images of 90 nm thick sections of S. pombe cells expressing Ost4-GFP-MTn (see Supplementary Chart 1 ).
    Paraformaldehyde Pfa, supplied by EM Science Inc, used in various techniques. Bioz Stars score: 92/100, based on 22 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/paraformaldehyde pfa/product/EM Science Inc
    Average 92 stars, based on 22 article reviews
    Price from $9.99 to $1999.99
    paraformaldehyde pfa - by Bioz Stars, 2020-08
    92/100 stars
      Buy from Supplier

    92
    Applichem paraformaldehyde pfa
    Strategies for achieving single-molecule visualization and cellular structural preservation in S. pombe cells. (a) The expression patterns of <t>Ost4-GFP,</t> Ost4-GFP-MTn, Nup124-GFP, Nup124-GFP-MTn, Sad1-GFP, and Sad1-GFP-MTn in S. pombe observed by live-cell fluorescence imaging (scale bar, 2 μm). The fluorescence images of GFP showed that the fusion proteins were correctly localized to expected subcellular locations: membranes of cortical ER and NE for Ost4, NPs for Nup124, the SPB for Sad1. (b) A cartoon shows the NE and cortical ER, NPs, and the SPB in a S. pombe cell. The regions of interest are labeled with numbers (I-VI) as references in other figures. (c) Procedures for the preparation of S. pombe cells for synthesizing AuNPs, three schemes labeled as 1 (“Cold MeOH”), 2 (“Oxidization + Chemical Fixation + Permeabilization”), 3 (“HPF/FSF + Oxidization + Chemical Fixation + Rehydration”) respectively, the Scheme 2 consists of four different sub-schemes (2a-2d), see the Supplementary Table 3 for details. (d) Synthesis of AuNPs in S. pombe cells treated by the “Cold MeOH” protocol with the optimized ANSM-based AuNPs synthesis protocol obtained with E. coli cells ( Fig.3b ) , the upper row shows the S. pombe cell suspension after AuNPs synthesis and the lower row shows the corresponding centrifuged cell pellets, apparently the colors of those MTn tags expressed cells are much darker than the corresponding control cells. (e) A typical EM image showing the unprecedented high labelling densities of AuNPs distributed on the cortical ER (II) and ER stacks (III) in a cell expressing Ost4-GFP-MTn prepared with the Scheme 1. (f) EM image showing better membranes preservation of an Ost4-GFP-MTn expressed cell treated with Scheme 2c than that in (e), the AuNPs could be clearly localized to outer membranes of the NE although the labeling densities of AuNPs reduced after the <t>PFA</t> fixation. (g) EM image showing the linear distributions of AuNPs along the membranes of cortical ER and ER stacks in a cell expressing Ost4-GFP-MTn treated with the Scheme 2a (with cold MeOH permeabilization), the structural preservation is better than that in (e) but with reduced labelling densities of AuNPs after the GA fixation. (h-i) EM images showing the AuNPs distributed along the membranes of NE (I) and ERs (II, IV) in cells expressing Ost4-GFP-MTn treated with Scheme 2b, lack of AuNPs in the nuclear pores (NPs), only a few background AuNPs scattered in cytosols, nuclei and mitochondria (M). (j) high densities of AuNPs formed on ER stacks in cells expressing Ost4-GFP-MTα treated with Scheme 2b (with 0.1% Triton X-100 2 min permeabilization). (k) A lot of AuNPs gathered in the SPB (among a dark density region), the specimen was prepared with Scheme 2a. (l) AuNPs synthesized in a cell expressing Nup124-GFP-MTn with Scheme 3a, grouped AuNPs formed on the NP (dark regions) with a regular interval along the NE. (m) Statistical analysis (one-way ANOVA followed by Brown-Forsythe test) of the AuNPs densities in ER (including NE), cytosol (Cyto), Nucleus (Nu) and mitochondrion (Mito) based on 20 EM images of 90 nm thick sections of S. pombe cells expressing Ost4-GFP-MTn (see Supplementary Chart 1 ).
    Paraformaldehyde Pfa, supplied by Applichem, used in various techniques. Bioz Stars score: 92/100, based on 40 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/paraformaldehyde pfa/product/Applichem
    Average 92 stars, based on 40 article reviews
    Price from $9.99 to $1999.99
    paraformaldehyde pfa - by Bioz Stars, 2020-08
    92/100 stars
      Buy from Supplier

    92
    Kodak Corp paraformaldehyde pfa
    Strategies for achieving single-molecule visualization and cellular structural preservation in S. pombe cells. (a) The expression patterns of <t>Ost4-GFP,</t> Ost4-GFP-MTn, Nup124-GFP, Nup124-GFP-MTn, Sad1-GFP, and Sad1-GFP-MTn in S. pombe observed by live-cell fluorescence imaging (scale bar, 2 μm). The fluorescence images of GFP showed that the fusion proteins were correctly localized to expected subcellular locations: membranes of cortical ER and NE for Ost4, NPs for Nup124, the SPB for Sad1. (b) A cartoon shows the NE and cortical ER, NPs, and the SPB in a S. pombe cell. The regions of interest are labeled with numbers (I-VI) as references in other figures. (c) Procedures for the preparation of S. pombe cells for synthesizing AuNPs, three schemes labeled as 1 (“Cold MeOH”), 2 (“Oxidization + Chemical Fixation + Permeabilization”), 3 (“HPF/FSF + Oxidization + Chemical Fixation + Rehydration”) respectively, the Scheme 2 consists of four different sub-schemes (2a-2d), see the Supplementary Table 3 for details. (d) Synthesis of AuNPs in S. pombe cells treated by the “Cold MeOH” protocol with the optimized ANSM-based AuNPs synthesis protocol obtained with E. coli cells ( Fig.3b ) , the upper row shows the S. pombe cell suspension after AuNPs synthesis and the lower row shows the corresponding centrifuged cell pellets, apparently the colors of those MTn tags expressed cells are much darker than the corresponding control cells. (e) A typical EM image showing the unprecedented high labelling densities of AuNPs distributed on the cortical ER (II) and ER stacks (III) in a cell expressing Ost4-GFP-MTn prepared with the Scheme 1. (f) EM image showing better membranes preservation of an Ost4-GFP-MTn expressed cell treated with Scheme 2c than that in (e), the AuNPs could be clearly localized to outer membranes of the NE although the labeling densities of AuNPs reduced after the <t>PFA</t> fixation. (g) EM image showing the linear distributions of AuNPs along the membranes of cortical ER and ER stacks in a cell expressing Ost4-GFP-MTn treated with the Scheme 2a (with cold MeOH permeabilization), the structural preservation is better than that in (e) but with reduced labelling densities of AuNPs after the GA fixation. (h-i) EM images showing the AuNPs distributed along the membranes of NE (I) and ERs (II, IV) in cells expressing Ost4-GFP-MTn treated with Scheme 2b, lack of AuNPs in the nuclear pores (NPs), only a few background AuNPs scattered in cytosols, nuclei and mitochondria (M). (j) high densities of AuNPs formed on ER stacks in cells expressing Ost4-GFP-MTα treated with Scheme 2b (with 0.1% Triton X-100 2 min permeabilization). (k) A lot of AuNPs gathered in the SPB (among a dark density region), the specimen was prepared with Scheme 2a. (l) AuNPs synthesized in a cell expressing Nup124-GFP-MTn with Scheme 3a, grouped AuNPs formed on the NP (dark regions) with a regular interval along the NE. (m) Statistical analysis (one-way ANOVA followed by Brown-Forsythe test) of the AuNPs densities in ER (including NE), cytosol (Cyto), Nucleus (Nu) and mitochondrion (Mito) based on 20 EM images of 90 nm thick sections of S. pombe cells expressing Ost4-GFP-MTn (see Supplementary Chart 1 ).
    Paraformaldehyde Pfa, supplied by Kodak Corp, used in various techniques. Bioz Stars score: 92/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/paraformaldehyde pfa/product/Kodak Corp
    Average 92 stars, based on 7 article reviews
    Price from $9.99 to $1999.99
    paraformaldehyde pfa - by Bioz Stars, 2020-08
    92/100 stars
      Buy from Supplier

    91
    Fisher Bioreagents paraformaldehyde pfa
    Strategies for achieving single-molecule visualization and cellular structural preservation in S. pombe cells. (a) The expression patterns of <t>Ost4-GFP,</t> Ost4-GFP-MTn, Nup124-GFP, Nup124-GFP-MTn, Sad1-GFP, and Sad1-GFP-MTn in S. pombe observed by live-cell fluorescence imaging (scale bar, 2 μm). The fluorescence images of GFP showed that the fusion proteins were correctly localized to expected subcellular locations: membranes of cortical ER and NE for Ost4, NPs for Nup124, the SPB for Sad1. (b) A cartoon shows the NE and cortical ER, NPs, and the SPB in a S. pombe cell. The regions of interest are labeled with numbers (I-VI) as references in other figures. (c) Procedures for the preparation of S. pombe cells for synthesizing AuNPs, three schemes labeled as 1 (“Cold MeOH”), 2 (“Oxidization + Chemical Fixation + Permeabilization”), 3 (“HPF/FSF + Oxidization + Chemical Fixation + Rehydration”) respectively, the Scheme 2 consists of four different sub-schemes (2a-2d), see the Supplementary Table 3 for details. (d) Synthesis of AuNPs in S. pombe cells treated by the “Cold MeOH” protocol with the optimized ANSM-based AuNPs synthesis protocol obtained with E. coli cells ( Fig.3b ) , the upper row shows the S. pombe cell suspension after AuNPs synthesis and the lower row shows the corresponding centrifuged cell pellets, apparently the colors of those MTn tags expressed cells are much darker than the corresponding control cells. (e) A typical EM image showing the unprecedented high labelling densities of AuNPs distributed on the cortical ER (II) and ER stacks (III) in a cell expressing Ost4-GFP-MTn prepared with the Scheme 1. (f) EM image showing better membranes preservation of an Ost4-GFP-MTn expressed cell treated with Scheme 2c than that in (e), the AuNPs could be clearly localized to outer membranes of the NE although the labeling densities of AuNPs reduced after the <t>PFA</t> fixation. (g) EM image showing the linear distributions of AuNPs along the membranes of cortical ER and ER stacks in a cell expressing Ost4-GFP-MTn treated with the Scheme 2a (with cold MeOH permeabilization), the structural preservation is better than that in (e) but with reduced labelling densities of AuNPs after the GA fixation. (h-i) EM images showing the AuNPs distributed along the membranes of NE (I) and ERs (II, IV) in cells expressing Ost4-GFP-MTn treated with Scheme 2b, lack of AuNPs in the nuclear pores (NPs), only a few background AuNPs scattered in cytosols, nuclei and mitochondria (M). (j) high densities of AuNPs formed on ER stacks in cells expressing Ost4-GFP-MTα treated with Scheme 2b (with 0.1% Triton X-100 2 min permeabilization). (k) A lot of AuNPs gathered in the SPB (among a dark density region), the specimen was prepared with Scheme 2a. (l) AuNPs synthesized in a cell expressing Nup124-GFP-MTn with Scheme 3a, grouped AuNPs formed on the NP (dark regions) with a regular interval along the NE. (m) Statistical analysis (one-way ANOVA followed by Brown-Forsythe test) of the AuNPs densities in ER (including NE), cytosol (Cyto), Nucleus (Nu) and mitochondrion (Mito) based on 20 EM images of 90 nm thick sections of S. pombe cells expressing Ost4-GFP-MTn (see Supplementary Chart 1 ).
    Paraformaldehyde Pfa, supplied by Fisher Bioreagents, used in various techniques. Bioz Stars score: 91/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/paraformaldehyde pfa/product/Fisher Bioreagents
    Average 91 stars, based on 5 article reviews
    Price from $9.99 to $1999.99
    paraformaldehyde pfa - by Bioz Stars, 2020-08
    91/100 stars
      Buy from Supplier

    93
    Becton Dickinson paraformaldehyde pfa
    Strategies for achieving single-molecule visualization and cellular structural preservation in S. pombe cells. (a) The expression patterns of <t>Ost4-GFP,</t> Ost4-GFP-MTn, Nup124-GFP, Nup124-GFP-MTn, Sad1-GFP, and Sad1-GFP-MTn in S. pombe observed by live-cell fluorescence imaging (scale bar, 2 μm). The fluorescence images of GFP showed that the fusion proteins were correctly localized to expected subcellular locations: membranes of cortical ER and NE for Ost4, NPs for Nup124, the SPB for Sad1. (b) A cartoon shows the NE and cortical ER, NPs, and the SPB in a S. pombe cell. The regions of interest are labeled with numbers (I-VI) as references in other figures. (c) Procedures for the preparation of S. pombe cells for synthesizing AuNPs, three schemes labeled as 1 (“Cold MeOH”), 2 (“Oxidization + Chemical Fixation + Permeabilization”), 3 (“HPF/FSF + Oxidization + Chemical Fixation + Rehydration”) respectively, the Scheme 2 consists of four different sub-schemes (2a-2d), see the Supplementary Table 3 for details. (d) Synthesis of AuNPs in S. pombe cells treated by the “Cold MeOH” protocol with the optimized ANSM-based AuNPs synthesis protocol obtained with E. coli cells ( Fig.3b ) , the upper row shows the S. pombe cell suspension after AuNPs synthesis and the lower row shows the corresponding centrifuged cell pellets, apparently the colors of those MTn tags expressed cells are much darker than the corresponding control cells. (e) A typical EM image showing the unprecedented high labelling densities of AuNPs distributed on the cortical ER (II) and ER stacks (III) in a cell expressing Ost4-GFP-MTn prepared with the Scheme 1. (f) EM image showing better membranes preservation of an Ost4-GFP-MTn expressed cell treated with Scheme 2c than that in (e), the AuNPs could be clearly localized to outer membranes of the NE although the labeling densities of AuNPs reduced after the <t>PFA</t> fixation. (g) EM image showing the linear distributions of AuNPs along the membranes of cortical ER and ER stacks in a cell expressing Ost4-GFP-MTn treated with the Scheme 2a (with cold MeOH permeabilization), the structural preservation is better than that in (e) but with reduced labelling densities of AuNPs after the GA fixation. (h-i) EM images showing the AuNPs distributed along the membranes of NE (I) and ERs (II, IV) in cells expressing Ost4-GFP-MTn treated with Scheme 2b, lack of AuNPs in the nuclear pores (NPs), only a few background AuNPs scattered in cytosols, nuclei and mitochondria (M). (j) high densities of AuNPs formed on ER stacks in cells expressing Ost4-GFP-MTα treated with Scheme 2b (with 0.1% Triton X-100 2 min permeabilization). (k) A lot of AuNPs gathered in the SPB (among a dark density region), the specimen was prepared with Scheme 2a. (l) AuNPs synthesized in a cell expressing Nup124-GFP-MTn with Scheme 3a, grouped AuNPs formed on the NP (dark regions) with a regular interval along the NE. (m) Statistical analysis (one-way ANOVA followed by Brown-Forsythe test) of the AuNPs densities in ER (including NE), cytosol (Cyto), Nucleus (Nu) and mitochondrion (Mito) based on 20 EM images of 90 nm thick sections of S. pombe cells expressing Ost4-GFP-MTn (see Supplementary Chart 1 ).
    Paraformaldehyde Pfa, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 93/100, based on 1527 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/paraformaldehyde pfa/product/Becton Dickinson
    Average 93 stars, based on 1527 article reviews
    Price from $9.99 to $1999.99
    paraformaldehyde pfa - by Bioz Stars, 2020-08
    93/100 stars
      Buy from Supplier

    93
    Shanghai Aladdin Bio-Chem paraformaldehyde pfa
    Strategies for achieving single-molecule visualization and cellular structural preservation in S. pombe cells. (a) The expression patterns of <t>Ost4-GFP,</t> Ost4-GFP-MTn, Nup124-GFP, Nup124-GFP-MTn, Sad1-GFP, and Sad1-GFP-MTn in S. pombe observed by live-cell fluorescence imaging (scale bar, 2 μm). The fluorescence images of GFP showed that the fusion proteins were correctly localized to expected subcellular locations: membranes of cortical ER and NE for Ost4, NPs for Nup124, the SPB for Sad1. (b) A cartoon shows the NE and cortical ER, NPs, and the SPB in a S. pombe cell. The regions of interest are labeled with numbers (I-VI) as references in other figures. (c) Procedures for the preparation of S. pombe cells for synthesizing AuNPs, three schemes labeled as 1 (“Cold MeOH”), 2 (“Oxidization + Chemical Fixation + Permeabilization”), 3 (“HPF/FSF + Oxidization + Chemical Fixation + Rehydration”) respectively, the Scheme 2 consists of four different sub-schemes (2a-2d), see the Supplementary Table 3 for details. (d) Synthesis of AuNPs in S. pombe cells treated by the “Cold MeOH” protocol with the optimized ANSM-based AuNPs synthesis protocol obtained with E. coli cells ( Fig.3b ) , the upper row shows the S. pombe cell suspension after AuNPs synthesis and the lower row shows the corresponding centrifuged cell pellets, apparently the colors of those MTn tags expressed cells are much darker than the corresponding control cells. (e) A typical EM image showing the unprecedented high labelling densities of AuNPs distributed on the cortical ER (II) and ER stacks (III) in a cell expressing Ost4-GFP-MTn prepared with the Scheme 1. (f) EM image showing better membranes preservation of an Ost4-GFP-MTn expressed cell treated with Scheme 2c than that in (e), the AuNPs could be clearly localized to outer membranes of the NE although the labeling densities of AuNPs reduced after the <t>PFA</t> fixation. (g) EM image showing the linear distributions of AuNPs along the membranes of cortical ER and ER stacks in a cell expressing Ost4-GFP-MTn treated with the Scheme 2a (with cold MeOH permeabilization), the structural preservation is better than that in (e) but with reduced labelling densities of AuNPs after the GA fixation. (h-i) EM images showing the AuNPs distributed along the membranes of NE (I) and ERs (II, IV) in cells expressing Ost4-GFP-MTn treated with Scheme 2b, lack of AuNPs in the nuclear pores (NPs), only a few background AuNPs scattered in cytosols, nuclei and mitochondria (M). (j) high densities of AuNPs formed on ER stacks in cells expressing Ost4-GFP-MTα treated with Scheme 2b (with 0.1% Triton X-100 2 min permeabilization). (k) A lot of AuNPs gathered in the SPB (among a dark density region), the specimen was prepared with Scheme 2a. (l) AuNPs synthesized in a cell expressing Nup124-GFP-MTn with Scheme 3a, grouped AuNPs formed on the NP (dark regions) with a regular interval along the NE. (m) Statistical analysis (one-way ANOVA followed by Brown-Forsythe test) of the AuNPs densities in ER (including NE), cytosol (Cyto), Nucleus (Nu) and mitochondrion (Mito) based on 20 EM images of 90 nm thick sections of S. pombe cells expressing Ost4-GFP-MTn (see Supplementary Chart 1 ).
    Paraformaldehyde Pfa, supplied by Shanghai Aladdin Bio-Chem, used in various techniques. Bioz Stars score: 93/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/paraformaldehyde pfa/product/Shanghai Aladdin Bio-Chem
    Average 93 stars, based on 3 article reviews
    Price from $9.99 to $1999.99
    paraformaldehyde pfa - by Bioz Stars, 2020-08
    93/100 stars
      Buy from Supplier

    93
    Cedarlane paraformaldehyde pfa
    Strategies for achieving single-molecule visualization and cellular structural preservation in S. pombe cells. (a) The expression patterns of <t>Ost4-GFP,</t> Ost4-GFP-MTn, Nup124-GFP, Nup124-GFP-MTn, Sad1-GFP, and Sad1-GFP-MTn in S. pombe observed by live-cell fluorescence imaging (scale bar, 2 μm). The fluorescence images of GFP showed that the fusion proteins were correctly localized to expected subcellular locations: membranes of cortical ER and NE for Ost4, NPs for Nup124, the SPB for Sad1. (b) A cartoon shows the NE and cortical ER, NPs, and the SPB in a S. pombe cell. The regions of interest are labeled with numbers (I-VI) as references in other figures. (c) Procedures for the preparation of S. pombe cells for synthesizing AuNPs, three schemes labeled as 1 (“Cold MeOH”), 2 (“Oxidization + Chemical Fixation + Permeabilization”), 3 (“HPF/FSF + Oxidization + Chemical Fixation + Rehydration”) respectively, the Scheme 2 consists of four different sub-schemes (2a-2d), see the Supplementary Table 3 for details. (d) Synthesis of AuNPs in S. pombe cells treated by the “Cold MeOH” protocol with the optimized ANSM-based AuNPs synthesis protocol obtained with E. coli cells ( Fig.3b ) , the upper row shows the S. pombe cell suspension after AuNPs synthesis and the lower row shows the corresponding centrifuged cell pellets, apparently the colors of those MTn tags expressed cells are much darker than the corresponding control cells. (e) A typical EM image showing the unprecedented high labelling densities of AuNPs distributed on the cortical ER (II) and ER stacks (III) in a cell expressing Ost4-GFP-MTn prepared with the Scheme 1. (f) EM image showing better membranes preservation of an Ost4-GFP-MTn expressed cell treated with Scheme 2c than that in (e), the AuNPs could be clearly localized to outer membranes of the NE although the labeling densities of AuNPs reduced after the <t>PFA</t> fixation. (g) EM image showing the linear distributions of AuNPs along the membranes of cortical ER and ER stacks in a cell expressing Ost4-GFP-MTn treated with the Scheme 2a (with cold MeOH permeabilization), the structural preservation is better than that in (e) but with reduced labelling densities of AuNPs after the GA fixation. (h-i) EM images showing the AuNPs distributed along the membranes of NE (I) and ERs (II, IV) in cells expressing Ost4-GFP-MTn treated with Scheme 2b, lack of AuNPs in the nuclear pores (NPs), only a few background AuNPs scattered in cytosols, nuclei and mitochondria (M). (j) high densities of AuNPs formed on ER stacks in cells expressing Ost4-GFP-MTα treated with Scheme 2b (with 0.1% Triton X-100 2 min permeabilization). (k) A lot of AuNPs gathered in the SPB (among a dark density region), the specimen was prepared with Scheme 2a. (l) AuNPs synthesized in a cell expressing Nup124-GFP-MTn with Scheme 3a, grouped AuNPs formed on the NP (dark regions) with a regular interval along the NE. (m) Statistical analysis (one-way ANOVA followed by Brown-Forsythe test) of the AuNPs densities in ER (including NE), cytosol (Cyto), Nucleus (Nu) and mitochondrion (Mito) based on 20 EM images of 90 nm thick sections of S. pombe cells expressing Ost4-GFP-MTn (see Supplementary Chart 1 ).
    Paraformaldehyde Pfa, supplied by Cedarlane, used in various techniques. Bioz Stars score: 93/100, based on 13 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/paraformaldehyde pfa/product/Cedarlane
    Average 93 stars, based on 13 article reviews
    Price from $9.99 to $1999.99
    paraformaldehyde pfa - by Bioz Stars, 2020-08
    93/100 stars
      Buy from Supplier

    Image Search Results


    Saccharides and saccharide-modifying enzymes do not affect N-CAM binding to knock-out progenitor cells. Cells were pretreated (1 hr) with heparin (0.01 mg/ml), chondroitin sulfate (0.01 mg/ml), heparinase (0.01 U/ml), or glycosidases (4 U/ml; N-glycosidase endo F and O-glycosidase) and then treated with N-CAM (10 μg/ml) for 2 hr at 37°C. The cells were washed with PBS, fixed with 4% PFA, and processed for immunocytochemistry for N-CAM, as described in Materials and Methods.

    Journal: The Journal of Neuroscience

    Article Title: N-CAM Binding Inhibits the Proliferation of Hippocampal Progenitor Cells and Promotes Their Differentiation to a Neuronal Phenotype

    doi: 10.1523/JNEUROSCI.20-10-03631.2000

    Figure Lengend Snippet: Saccharides and saccharide-modifying enzymes do not affect N-CAM binding to knock-out progenitor cells. Cells were pretreated (1 hr) with heparin (0.01 mg/ml), chondroitin sulfate (0.01 mg/ml), heparinase (0.01 U/ml), or glycosidases (4 U/ml; N-glycosidase endo F and O-glycosidase) and then treated with N-CAM (10 μg/ml) for 2 hr at 37°C. The cells were washed with PBS, fixed with 4% PFA, and processed for immunocytochemistry for N-CAM, as described in Materials and Methods.

    Article Snippet: After 48 hr of treatment with N-CAM (10 μg/ml), cells were fixed with 4% PFA (Sigma).

    Techniques: Chick Chorioallantoic Membrane Assay, Binding Assay, Knock-Out, Immunocytochemistry

    Binding of N-CAM and extracellular N-CAM to live hippocampal progenitor cells from N-CAM knock-out mice. Cells were treated with N-CAM, the extracellular domain of N-CAM, and the recombinant Ig III domain (10 μg/ml) for 2 hr at 37°C. The cells were washed with PBS, fixed with 4% PFA, and processed for immunocytochemistry using antibodies for N-CAM and N-CAM domains and FITC-labeled secondary antibodies as described in Materials and Methods. Nuclei were revealed by counterstaining with DAPI.

    Journal: The Journal of Neuroscience

    Article Title: N-CAM Binding Inhibits the Proliferation of Hippocampal Progenitor Cells and Promotes Their Differentiation to a Neuronal Phenotype

    doi: 10.1523/JNEUROSCI.20-10-03631.2000

    Figure Lengend Snippet: Binding of N-CAM and extracellular N-CAM to live hippocampal progenitor cells from N-CAM knock-out mice. Cells were treated with N-CAM, the extracellular domain of N-CAM, and the recombinant Ig III domain (10 μg/ml) for 2 hr at 37°C. The cells were washed with PBS, fixed with 4% PFA, and processed for immunocytochemistry using antibodies for N-CAM and N-CAM domains and FITC-labeled secondary antibodies as described in Materials and Methods. Nuclei were revealed by counterstaining with DAPI.

    Article Snippet: After 48 hr of treatment with N-CAM (10 μg/ml), cells were fixed with 4% PFA (Sigma).

    Techniques: Binding Assay, Chick Chorioallantoic Membrane Assay, Knock-Out, Mouse Assay, Recombinant, Immunocytochemistry, Labeling

    Protocol for 3D blood vessel visualization in whole mouse brains. (a) Sample preparation: During animal sacrifice mice are transcardially perfused with 40 mL of phosphate-buffered saline (PBS) containing 100 U/mL heparin, followed by 20 mL of 4% paraformaldehyde (PFA) in PBS and 10 mL hydrogel composed of 2% gelatin and 0.1% FITC-conjugated albumin in PBS. (b) Whole brain clearing: In the optimized protocol, the brain specimen is incubated in each clearing solution (tetrahydrofuran (THF) used for dehydration and lipid solvation, dibenzyl ether (DBE) for matching of refractive index) for 12 h at room temperature with constant shaking in dark bottles. To ensure complete dehydration, the 100% THF step was performed twice. (c) Light sheet microscopy: Cleared brains immersed in DBE were imaged in a dorso-ventral direction using either 1.6 × or 6.4 × magnification and corresponding step sizes of 6 µm and 4 µm in the LaVision Biotec ultramicroscope.

    Journal: Journal of Cerebral Blood Flow & Metabolism

    Article Title: 3D visualization and quantification of microvessels in the whole ischemic mouse brain using solvent-based clearing and light sheet microscopy

    doi: 10.1177/0271678X17698970

    Figure Lengend Snippet: Protocol for 3D blood vessel visualization in whole mouse brains. (a) Sample preparation: During animal sacrifice mice are transcardially perfused with 40 mL of phosphate-buffered saline (PBS) containing 100 U/mL heparin, followed by 20 mL of 4% paraformaldehyde (PFA) in PBS and 10 mL hydrogel composed of 2% gelatin and 0.1% FITC-conjugated albumin in PBS. (b) Whole brain clearing: In the optimized protocol, the brain specimen is incubated in each clearing solution (tetrahydrofuran (THF) used for dehydration and lipid solvation, dibenzyl ether (DBE) for matching of refractive index) for 12 h at room temperature with constant shaking in dark bottles. To ensure complete dehydration, the 100% THF step was performed twice. (c) Light sheet microscopy: Cleared brains immersed in DBE were imaged in a dorso-ventral direction using either 1.6 × or 6.4 × magnification and corresponding step sizes of 6 µm and 4 µm in the LaVision Biotec ultramicroscope.

    Article Snippet: Mice were deeply anesthetized by an intraperitoneal injection of 7% chloral hydrate and transcardially perfused with 40 mL of PBS containing 50 U/mL heparin (Ratiopharm, Ulm, Germany), followed by perfusion of 20 mL of 4% paraformaldehyde (PFA) (Merck-Millipore) in PBS.

    Techniques: Sample Prep, Mouse Assay, Incubation, Microscopy

    Strategies for achieving single-molecule visualization and cellular structural preservation in S. pombe cells. (a) The expression patterns of Ost4-GFP, Ost4-GFP-MTn, Nup124-GFP, Nup124-GFP-MTn, Sad1-GFP, and Sad1-GFP-MTn in S. pombe observed by live-cell fluorescence imaging (scale bar, 2 μm). The fluorescence images of GFP showed that the fusion proteins were correctly localized to expected subcellular locations: membranes of cortical ER and NE for Ost4, NPs for Nup124, the SPB for Sad1. (b) A cartoon shows the NE and cortical ER, NPs, and the SPB in a S. pombe cell. The regions of interest are labeled with numbers (I-VI) as references in other figures. (c) Procedures for the preparation of S. pombe cells for synthesizing AuNPs, three schemes labeled as 1 (“Cold MeOH”), 2 (“Oxidization + Chemical Fixation + Permeabilization”), 3 (“HPF/FSF + Oxidization + Chemical Fixation + Rehydration”) respectively, the Scheme 2 consists of four different sub-schemes (2a-2d), see the Supplementary Table 3 for details. (d) Synthesis of AuNPs in S. pombe cells treated by the “Cold MeOH” protocol with the optimized ANSM-based AuNPs synthesis protocol obtained with E. coli cells ( Fig.3b ) , the upper row shows the S. pombe cell suspension after AuNPs synthesis and the lower row shows the corresponding centrifuged cell pellets, apparently the colors of those MTn tags expressed cells are much darker than the corresponding control cells. (e) A typical EM image showing the unprecedented high labelling densities of AuNPs distributed on the cortical ER (II) and ER stacks (III) in a cell expressing Ost4-GFP-MTn prepared with the Scheme 1. (f) EM image showing better membranes preservation of an Ost4-GFP-MTn expressed cell treated with Scheme 2c than that in (e), the AuNPs could be clearly localized to outer membranes of the NE although the labeling densities of AuNPs reduced after the PFA fixation. (g) EM image showing the linear distributions of AuNPs along the membranes of cortical ER and ER stacks in a cell expressing Ost4-GFP-MTn treated with the Scheme 2a (with cold MeOH permeabilization), the structural preservation is better than that in (e) but with reduced labelling densities of AuNPs after the GA fixation. (h-i) EM images showing the AuNPs distributed along the membranes of NE (I) and ERs (II, IV) in cells expressing Ost4-GFP-MTn treated with Scheme 2b, lack of AuNPs in the nuclear pores (NPs), only a few background AuNPs scattered in cytosols, nuclei and mitochondria (M). (j) high densities of AuNPs formed on ER stacks in cells expressing Ost4-GFP-MTα treated with Scheme 2b (with 0.1% Triton X-100 2 min permeabilization). (k) A lot of AuNPs gathered in the SPB (among a dark density region), the specimen was prepared with Scheme 2a. (l) AuNPs synthesized in a cell expressing Nup124-GFP-MTn with Scheme 3a, grouped AuNPs formed on the NP (dark regions) with a regular interval along the NE. (m) Statistical analysis (one-way ANOVA followed by Brown-Forsythe test) of the AuNPs densities in ER (including NE), cytosol (Cyto), Nucleus (Nu) and mitochondrion (Mito) based on 20 EM images of 90 nm thick sections of S. pombe cells expressing Ost4-GFP-MTn (see Supplementary Chart 1 ).

    Journal: bioRxiv

    Article Title: Direct synthesis of EM-visible gold nanoparticles on genetically encoded tags for single-molecule visualization in cells

    doi: 10.1101/520999

    Figure Lengend Snippet: Strategies for achieving single-molecule visualization and cellular structural preservation in S. pombe cells. (a) The expression patterns of Ost4-GFP, Ost4-GFP-MTn, Nup124-GFP, Nup124-GFP-MTn, Sad1-GFP, and Sad1-GFP-MTn in S. pombe observed by live-cell fluorescence imaging (scale bar, 2 μm). The fluorescence images of GFP showed that the fusion proteins were correctly localized to expected subcellular locations: membranes of cortical ER and NE for Ost4, NPs for Nup124, the SPB for Sad1. (b) A cartoon shows the NE and cortical ER, NPs, and the SPB in a S. pombe cell. The regions of interest are labeled with numbers (I-VI) as references in other figures. (c) Procedures for the preparation of S. pombe cells for synthesizing AuNPs, three schemes labeled as 1 (“Cold MeOH”), 2 (“Oxidization + Chemical Fixation + Permeabilization”), 3 (“HPF/FSF + Oxidization + Chemical Fixation + Rehydration”) respectively, the Scheme 2 consists of four different sub-schemes (2a-2d), see the Supplementary Table 3 for details. (d) Synthesis of AuNPs in S. pombe cells treated by the “Cold MeOH” protocol with the optimized ANSM-based AuNPs synthesis protocol obtained with E. coli cells ( Fig.3b ) , the upper row shows the S. pombe cell suspension after AuNPs synthesis and the lower row shows the corresponding centrifuged cell pellets, apparently the colors of those MTn tags expressed cells are much darker than the corresponding control cells. (e) A typical EM image showing the unprecedented high labelling densities of AuNPs distributed on the cortical ER (II) and ER stacks (III) in a cell expressing Ost4-GFP-MTn prepared with the Scheme 1. (f) EM image showing better membranes preservation of an Ost4-GFP-MTn expressed cell treated with Scheme 2c than that in (e), the AuNPs could be clearly localized to outer membranes of the NE although the labeling densities of AuNPs reduced after the PFA fixation. (g) EM image showing the linear distributions of AuNPs along the membranes of cortical ER and ER stacks in a cell expressing Ost4-GFP-MTn treated with the Scheme 2a (with cold MeOH permeabilization), the structural preservation is better than that in (e) but with reduced labelling densities of AuNPs after the GA fixation. (h-i) EM images showing the AuNPs distributed along the membranes of NE (I) and ERs (II, IV) in cells expressing Ost4-GFP-MTn treated with Scheme 2b, lack of AuNPs in the nuclear pores (NPs), only a few background AuNPs scattered in cytosols, nuclei and mitochondria (M). (j) high densities of AuNPs formed on ER stacks in cells expressing Ost4-GFP-MTα treated with Scheme 2b (with 0.1% Triton X-100 2 min permeabilization). (k) A lot of AuNPs gathered in the SPB (among a dark density region), the specimen was prepared with Scheme 2a. (l) AuNPs synthesized in a cell expressing Nup124-GFP-MTn with Scheme 3a, grouped AuNPs formed on the NP (dark regions) with a regular interval along the NE. (m) Statistical analysis (one-way ANOVA followed by Brown-Forsythe test) of the AuNPs densities in ER (including NE), cytosol (Cyto), Nucleus (Nu) and mitochondrion (Mito) based on 20 EM images of 90 nm thick sections of S. pombe cells expressing Ost4-GFP-MTn (see Supplementary Chart 1 ).

    Article Snippet: For comparison, another commonly used fixative, paraformaldehyde (PFA) (Cat.A11313, Alfa Aesar), was tested with Ost4-GFP-MTn expressing S. pombe cells by replacing the 0.5% GA with 4% PFA and 3 or 5mM DTDPA (freshly prepared) in 100 mM PIPES buffer with 1 mM MgCl2 , 1 mM CaCl2 and 0.1 M sorbitol (pH 6.8) for 1 h fixation at 4 °C.

    Techniques: Preserving, Expressing, Fluorescence, Imaging, Labeling, Synthesized