triethylammonium Thermo Fisher Search Results


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  • 95
    Millipore triethyl ammonium bicarbonate buffer teab
    Triethyl Ammonium Bicarbonate Buffer Teab, supplied by Millipore, used in various techniques. Bioz Stars score: 95/100, based on 16 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Thermo Fisher alexa fluor 488 azide
    Reaction scheme for creating click-conjugated dendrons employed in the in vitro studies. (i) glutaric anhydride, MeOH, 24 hrs, r.t. (ii) EDC, c(RGDyK), H 2 O/DMSO, 2 days, r.t. (iii) <t>Alexa</t> <t>Fluor</t> 488 azide OR biotin-dPEG 3+4 -azide OR MTX - azide (5) OP azide-G2(AF)
    Alexa Fluor 488 Azide, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 702 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    84
    Thermo Fisher triethylammonium salt tritc dhpe
    Analysis of domain-specific <t>TRITC-DHPE</t> diffusivity ( A ) and E raft data of NBD-DHPE distribution ( B ) suggest that variation in membrane CHOL between 25 and 37 mol% CHOL alters lipid packing differences between l o and l d domains of a polymer-tethered lipid bilayer containing DOPC/DPPC/CHOL mixtures (1:1 DOPC/DPPC molar ratio); p
    Triethylammonium Salt Tritc Dhpe, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 84/100, based on 14 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    78
    Thermo Fisher tris triethylammonium salt
    Analysis of domain-specific <t>TRITC-DHPE</t> diffusivity ( A ) and E raft data of NBD-DHPE distribution ( B ) suggest that variation in membrane CHOL between 25 and 37 mol% CHOL alters lipid packing differences between l o and l d domains of a polymer-tethered lipid bilayer containing DOPC/DPPC/CHOL mixtures (1:1 DOPC/DPPC molar ratio); p
    Tris Triethylammonium Salt, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 78/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Thermo Fisher triethylammonium bicarbonate
    Analysis of domain-specific <t>TRITC-DHPE</t> diffusivity ( A ) and E raft data of NBD-DHPE distribution ( B ) suggest that variation in membrane CHOL between 25 and 37 mol% CHOL alters lipid packing differences between l o and l d domains of a polymer-tethered lipid bilayer containing DOPC/DPPC/CHOL mixtures (1:1 DOPC/DPPC molar ratio); p
    Triethylammonium Bicarbonate, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 202 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Thermo Fisher fm2 10
    Imaging of enhanced release with <t>FM2-10</t> styryl dye. A , NMJs were first loaded with FM2-10 by stimulating 90 s at 10 Hz and then challenged (separately) with different stimulation trains. B , In response to 120 s at 50 Hz stimulation train, both WT and R6/1 synapses lost the FM2-10 dye almost completely. No obvious changes in destaining kinetics were observed between WT and R6/1. C , D , Upon milder stimulation with 40 s at 10 Hz trains, mutant synapses destained more and faster than WT synapses, as expected for higher release probability at R6/1 synapses. E , Recycling pool recovery from depletion (induced by a 30 s at 100 Hz train) was assessed by measuring EPP amplitude evoked by sequential stimulation at 2 Hz. Although the depression is stronger for R6/1 synapses (for clarity, only 1 point of 10 is drawn), the recovery was very similar in WT and mutant synapses. Thus, no obvious abnormalities occurred in the refilling of the recycling pool at R6/1 synapses.
    Fm2 10, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 93/100, based on 114 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    84
    Thermo Fisher triethylammonium acetate
    Imaging of enhanced release with <t>FM2-10</t> styryl dye. A , NMJs were first loaded with FM2-10 by stimulating 90 s at 10 Hz and then challenged (separately) with different stimulation trains. B , In response to 120 s at 50 Hz stimulation train, both WT and R6/1 synapses lost the FM2-10 dye almost completely. No obvious changes in destaining kinetics were observed between WT and R6/1. C , D , Upon milder stimulation with 40 s at 10 Hz trains, mutant synapses destained more and faster than WT synapses, as expected for higher release probability at R6/1 synapses. E , Recycling pool recovery from depletion (induced by a 30 s at 100 Hz train) was assessed by measuring EPP amplitude evoked by sequential stimulation at 2 Hz. Although the depression is stronger for R6/1 synapses (for clarity, only 1 point of 10 is drawn), the recovery was very similar in WT and mutant synapses. Thus, no obvious abnormalities occurred in the refilling of the recycling pool at R6/1 synapses.
    Triethylammonium Acetate, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 84/100, based on 19 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    77
    Thermo Fisher lipid dye fm 1 43
    The status of the membranous organelles (MOs) in sperm. A) When labeled with the membrane dye FM® 1–43 (Life Technologies™), fused MOs are visible as bright spots just inside the cell membrane, but unfused MOs are not labeled. Here, the sperm are shown in both DIC illumination and epifluorescence illumination. Top panel: a spermatozoon with an obvious pseudopod and the bright foci in the cell body indicating fused MOs. Middle panel: an abnormal spermatid with fused MOs. Bottom panel: a normal spermatid with no fused MOs. B) The percentage of normal spermatids and those that had at least a single obvious fused MO from  him-5  and  spe-46; him-5  male worms. Error bars represent SEM. In C-F, the sperm have been fixed, permeabilized, and exposed to an Alexa Fluor® 594 conjugate of wheat germ agglutinin (WGA; Life Technologies™), which labels all MOs, including those that have not fused with the cell membrane (original red fluorescence false colored green). C) Sperm from a  spe-46(hc197); him-5(e1490)  male. D) The same cells as is C, visualized in DIC optics. E) Sperm from a  him-5(e1490)  male labeled as in C. F) DIC image of the cells in E.
    Lipid Dye Fm 1 43, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 77/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    79
    Thermo Fisher cationic styrylpyridinium dye fm 1 43
    The status of the membranous organelles (MOs) in sperm. A) When labeled with the membrane dye FM® 1–43 (Life Technologies™), fused MOs are visible as bright spots just inside the cell membrane, but unfused MOs are not labeled. Here, the sperm are shown in both DIC illumination and epifluorescence illumination. Top panel: a spermatozoon with an obvious pseudopod and the bright foci in the cell body indicating fused MOs. Middle panel: an abnormal spermatid with fused MOs. Bottom panel: a normal spermatid with no fused MOs. B) The percentage of normal spermatids and those that had at least a single obvious fused MO from  him-5  and  spe-46; him-5  male worms. Error bars represent SEM. In C-F, the sperm have been fixed, permeabilized, and exposed to an Alexa Fluor® 594 conjugate of wheat germ agglutinin (WGA; Life Technologies™), which labels all MOs, including those that have not fused with the cell membrane (original red fluorescence false colored green). C) Sperm from a  spe-46(hc197); him-5(e1490)  male. D) The same cells as is C, visualized in DIC optics. E) Sperm from a  him-5(e1490)  male labeled as in C. F) DIC image of the cells in E.
    Cationic Styrylpyridinium Dye Fm 1 43, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 79/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    76
    Thermo Fisher triethylammonium salt rh pe
    The status of the membranous organelles (MOs) in sperm. A) When labeled with the membrane dye FM® 1–43 (Life Technologies™), fused MOs are visible as bright spots just inside the cell membrane, but unfused MOs are not labeled. Here, the sperm are shown in both DIC illumination and epifluorescence illumination. Top panel: a spermatozoon with an obvious pseudopod and the bright foci in the cell body indicating fused MOs. Middle panel: an abnormal spermatid with fused MOs. Bottom panel: a normal spermatid with no fused MOs. B) The percentage of normal spermatids and those that had at least a single obvious fused MO from  him-5  and  spe-46; him-5  male worms. Error bars represent SEM. In C-F, the sperm have been fixed, permeabilized, and exposed to an Alexa Fluor® 594 conjugate of wheat germ agglutinin (WGA; Life Technologies™), which labels all MOs, including those that have not fused with the cell membrane (original red fluorescence false colored green). C) Sperm from a  spe-46(hc197); him-5(e1490)  male. D) The same cells as is C, visualized in DIC optics. E) Sperm from a  him-5(e1490)  male labeled as in C. F) DIC image of the cells in E.
    Triethylammonium Salt Rh Pe, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 76/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    87
    Thermo Fisher itraq kit
    The status of the membranous organelles (MOs) in sperm. A) When labeled with the membrane dye FM® 1–43 (Life Technologies™), fused MOs are visible as bright spots just inside the cell membrane, but unfused MOs are not labeled. Here, the sperm are shown in both DIC illumination and epifluorescence illumination. Top panel: a spermatozoon with an obvious pseudopod and the bright foci in the cell body indicating fused MOs. Middle panel: an abnormal spermatid with fused MOs. Bottom panel: a normal spermatid with no fused MOs. B) The percentage of normal spermatids and those that had at least a single obvious fused MO from  him-5  and  spe-46; him-5  male worms. Error bars represent SEM. In C-F, the sperm have been fixed, permeabilized, and exposed to an Alexa Fluor® 594 conjugate of wheat germ agglutinin (WGA; Life Technologies™), which labels all MOs, including those that have not fused with the cell membrane (original red fluorescence false colored green). C) Sperm from a  spe-46(hc197); him-5(e1490)  male. D) The same cells as is C, visualized in DIC optics. E) Sperm from a  him-5(e1490)  male labeled as in C. F) DIC image of the cells in E.
    Itraq Kit, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 87/100, based on 43 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    78
    Thermo Fisher endocytic marker fm 1 43
    The PLD antagonist  n -ButOH and DGK inhibitor R59022 disrupt internalization of the endomembrane tracer dye FM 1-43 .  (A)  Ninety-minute-old pollen tubes were pre-incubated in germination medium on ice for 10 min, and 2 μM FM1-43 dye together with a particular drug or PA were then simultaneously added and the cells monitored for 30 min. Representative pollen tubes observed at two times are shown.  (B)  Alternatively, pollen tubes were incubated in germination medium containing 2 μM FM1-43 for 90 min, a particular drug or PA were then added for 10 min and the cells were observed with a confocal microscope. At least 15 cells were analyzed for each treatment in four independent experiments and typical examples are shown. Scale bar = 10 μm.
    Endocytic Marker Fm 1 43, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 78/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Thermo Fisher filmtracer fm 1 43 green biofilm dye
    CRISPRi-mediated phenotypes associated with <t>biofilm</t> structure and EPS matrix. P . fluorescens SBW25 cells expressing the CRISPRi system targeting genes gacS (PFLU3777, two-component sensor kinase), rimA (PFLU0263, PDE), dipA (PFLU0458, PDE), alg44 (PFLU0988, alginase co-polymerase Alg44), gcbA (PFL0621, DGC) and bifA (PFLU4858, PDE). The control corresponds to the pPFL-gRNA plasmid with no guide RNA inserted. (A) Cells were stained with FM 1–43 and observed by confocal microscopy. 3D architecture of biofilms were reconstructed as described in Methods. Virtual shadow projections were included on the right to show thickness. (B) Biofilms grown in the presence of the Congo Red dye to reveal the structure of EPS. The ESP-stained volumes were rendered with maximum intensity projection. Inset shows a blow-up view of a clump in the EPS matrix after silencing of bifA .
    Filmtracer Fm 1 43 Green Biofilm Dye, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 85/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    81
    Thermo Fisher tritc dhpe
    Bilayer calibration curves and calculation of binding ratio. ( A ) Intensity of NBD-PC bilayer at different molecular densities with the F factor comparing brightness of Notch1-eGFP and NBD-PC. ( B ) Calibration curve and F factor relating <t>TRITC-DHPE</t> and
    Tritc Dhpe, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 81/100, based on 22 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    78
    Thermo Fisher n 3 triethylammonium propyl 4 dibutilamino styrylpyrodinum dibromide fm1 43
    Bilayer calibration curves and calculation of binding ratio. ( A ) Intensity of NBD-PC bilayer at different molecular densities with the F factor comparing brightness of Notch1-eGFP and NBD-PC. ( B ) Calibration curve and F factor relating <t>TRITC-DHPE</t> and
    N 3 Triethylammonium Propyl 4 Dibutilamino Styrylpyrodinum Dibromide Fm1 43, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 78/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Thermo Fisher alexa fluor 594
    Generation of antioxidant over-expressing parasites. Transgenic parasites were created in the RHΔhxgprt background strain using plasmids designed to over-express HA-tagged versions of glutaredoxin (Glut), catalase (Cat), or peroxiredoxin 2 (PRx2) under the control of the gra1 gene promoter. ( a ) Over-expression was verified using quantitative real-time PCR to measure transcript levels. ( b ) Western blot analysis was used to confirm protein production and that the resultant proteins were the appropriate size. ( c ) Immunofluorescence assays were performed for protein localization, utilizing a rabbit monoclonal antibody against the HA tag and an <t>Alexa</t> <t>Fluor</t> 594 secondary antibody. ( d ) General parasite replication was determined by doubling assay. Confluent HFF in 24-well plates were infected with the indicated parasite lines for 5 hours before the medium was replaced with fresh growth medium. Parasites were allowed to grow for 32 hours before being methanol-fixed and stained with DiffQuick. The number of individual parasites per vacuole was enumerated for at least 50 vacuoles. Data is presented as percent of vacuoles from 4 experiments. Student’s t-test was employed for statistical analysis. Scale bars equal 2 μm.
    Alexa Fluor 594, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 13743 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    75
    Thermo Fisher alexa fluor 594 alkyne alexa fluor 594 carboxamido 5 and 6 propargyl bis triethylammonium salt
    Images of immunofluorescence by confocal laser scanning microscopy (CLSM) of four different breast cancer cell lines: MCF-7, SKBR3, T47D and MDA-MB-231 ( a ) bright field; ( b ) staining of the cell nucleus by the fluorophore DAPI (blue); ( c ) indirect detection of HER2 by a primary anti-HER2 antibody and an anti-anti-HER2 secondary antibody bound to <t>Alexa</t> <t>Fluor</t> 594 fluorophore (red) and ( d ) cell localization of HER2 receptor by overlapping of images (DAPI + HER2). Magnification: 63×.
    Alexa Fluor 594 Alkyne Alexa Fluor 594 Carboxamido 5 And 6 Propargyl Bis Triethylammonium Salt, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 75/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    75
    Thermo Fisher triethylammonium salt dansyl
    Images of immunofluorescence by confocal laser scanning microscopy (CLSM) of four different breast cancer cell lines: MCF-7, SKBR3, T47D and MDA-MB-231 ( a ) bright field; ( b ) staining of the cell nucleus by the fluorophore DAPI (blue); ( c ) indirect detection of HER2 by a primary anti-HER2 antibody and an anti-anti-HER2 secondary antibody bound to <t>Alexa</t> <t>Fluor</t> 594 fluorophore (red) and ( d ) cell localization of HER2 receptor by overlapping of images (DAPI + HER2). Magnification: 63×.
    Triethylammonium Salt Dansyl, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 75/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    76
    Thermo Fisher triethylammonium salt tr dppe
    Images of immunofluorescence by confocal laser scanning microscopy (CLSM) of four different breast cancer cell lines: MCF-7, SKBR3, T47D and MDA-MB-231 ( a ) bright field; ( b ) staining of the cell nucleus by the fluorophore DAPI (blue); ( c ) indirect detection of HER2 by a primary anti-HER2 antibody and an anti-anti-HER2 secondary antibody bound to <t>Alexa</t> <t>Fluor</t> 594 fluorophore (red) and ( d ) cell localization of HER2 receptor by overlapping of images (DAPI + HER2). Magnification: 63×.
    Triethylammonium Salt Tr Dppe, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 76/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Thermo Fisher nbd pe
    Time-lapse fluorescence microscopy images of shrinking vesicles. (a) The GFP leaked from DOPC/ 5 GUV upon the shrinkage of the vesicle by PLA 2 treatment. The GUV membrane was stained with Texas Red <t>DHPE.</t> (b) The 100 nm red fluorescent sulfate-modified polystyrene nanoparticles were contained and concentrated in DOPC/ 5 GUV as the vesicle shrank. The membrane was stained with <t>NBD-PE.</t>
    Nbd Pe, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 70 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher alexafluor 488
    (a) Flow analysis to show aggregated human γ-globulin (AHG)–AlexaFluor® 488 binding to CD4 +  T cells in purified peripheral blood mononuclear cells (PBMCs). Cells were gated using forward- and side-scatter. CD4 +  lymphocytes were
    Alexafluor 488, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 6999 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher oregon green 488 taxol
    Comparison of OATP1B1- and OATP1B3-mediated transport of fluorescent substrates. Uptake was measured with 1 µM of the indicated fluorescent compounds on 24-well plates for 30 min. Values are means ± SE of two combined independent experiments each performed with triplicate determinations. FMTX: fluorescein methotrexate; AMTX: Alexa Fluor® 488 methotrexate; Flutax-2: Oregon Green® 488 <t>Taxol;</t> WT: wild-type CHO cells; OATP1B1: CHO cells expressing OATP1B1; OATP1B3: CHO cells expressing OATP1B3.
    Oregon Green 488 Taxol, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher tris
    Comparison of OATP1B1- and OATP1B3-mediated transport of fluorescent substrates. Uptake was measured with 1 µM of the indicated fluorescent compounds on 24-well plates for 30 min. Values are means ± SE of two combined independent experiments each performed with triplicate determinations. FMTX: fluorescein methotrexate; AMTX: Alexa Fluor® 488 methotrexate; Flutax-2: Oregon Green® 488 <t>Taxol;</t> WT: wild-type CHO cells; OATP1B1: CHO cells expressing OATP1B1; OATP1B3: CHO cells expressing OATP1B3.
    Tris, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 11798 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    79
    Thermo Fisher membrane stain fm 1 43
    Characterization of biogenic Mn oxides produced by strain T-G1 in PYG medium at pH 5.5. (A)  Scanning electron micrograph showing spherical clusters of micron to sub-micron sheets of Mn oxide minerals.  (B)  An enlargement of the area indicated by a red box in  (A) .  (C)  Confocal laser scanning microscopy (CLSM) of a mineral aggregate in association with T-G1. Bacterial cells stained by FM 1-43 probe shown in green, reflection of minerals was indicated in gray.  (D)  Energy-dispersive X-ray (EDS) spectra showing elemental composition of biogenic Mn oxide particles.  (E)  Raman spectroscopy of biogenic Mn oxides.  (F)  XRD pattern of biogenic Mn-oxide minerals with vertical lines indicating strong (red) and weak peak positions (green) of bixbyite.
    Membrane Stain Fm 1 43, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 79/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    79
    Thermo Fisher bulk plasma membrane marker fm 1 43
    Characterization of biogenic Mn oxides produced by strain T-G1 in PYG medium at pH 5.5. (A)  Scanning electron micrograph showing spherical clusters of micron to sub-micron sheets of Mn oxide minerals.  (B)  An enlargement of the area indicated by a red box in  (A) .  (C)  Confocal laser scanning microscopy (CLSM) of a mineral aggregate in association with T-G1. Bacterial cells stained by FM 1-43 probe shown in green, reflection of minerals was indicated in gray.  (D)  Energy-dispersive X-ray (EDS) spectra showing elemental composition of biogenic Mn oxide particles.  (E)  Raman spectroscopy of biogenic Mn oxides.  (F)  XRD pattern of biogenic Mn-oxide minerals with vertical lines indicating strong (red) and weak peak positions (green) of bixbyite.
    Bulk Plasma Membrane Marker Fm 1 43, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 79/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher dye rh414 n 3 triethylammonium propyl 4 4 p diethylaminophenyl butadienyl pyridinium dibromide
    Characterization of biogenic Mn oxides produced by strain T-G1 in PYG medium at pH 5.5. (A)  Scanning electron micrograph showing spherical clusters of micron to sub-micron sheets of Mn oxide minerals.  (B)  An enlargement of the area indicated by a red box in  (A) .  (C)  Confocal laser scanning microscopy (CLSM) of a mineral aggregate in association with T-G1. Bacterial cells stained by FM 1-43 probe shown in green, reflection of minerals was indicated in gray.  (D)  Energy-dispersive X-ray (EDS) spectra showing elemental composition of biogenic Mn oxide particles.  (E)  Raman spectroscopy of biogenic Mn oxides.  (F)  XRD pattern of biogenic Mn-oxide minerals with vertical lines indicating strong (red) and weak peak positions (green) of bixbyite.
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    Thermo Fisher cell stain fm 1 43
    Intracellular calcium responses of trigeminal sensory neurons innervating the cornea of control and 4-week tear-deficient guinea pigs. (A) Trigeminal ganglion (TG) neurons innervating the cornea were retrogradely labeled with FM 1-43 applied on the corneal surface 6 days earlier and identified with fluorescence microscopy. Only one or 2 corneal neurons were found per culture plate. Photographs show (from left to right) bright field, FM 1-43 fluorescence when excited with 470-nm light and merged images. (B–E) Ratiometric fluorescence changes (expressed as the ratio of fluorescence of Fura-2 when excited at 340 and 380 nm, F 340 /F 380 ) recorded simultaneously in the same culture plate from several fresh cultured TG sensory neurons from control (B and D) and tear-deficient (C and E) animals, in response to a cooling ramp and to the addition to the bath solution of 100 μM menthol, 100 μM cinnamaldehyde, 1 μM capsaicin, and 30 mM K + ; the bath temperature was simultaneously recorded (lower channels). The blue lines correspond to recordings from TG neurons innervating the cornea. Neurons responding to cooling and menthol were classified as cold thermosensitive neurons (B and C); neurons not responding to cooling stimulus but responding to capsaicin and cinnamaldehyde were classified as putative polymodal nociceptive neurons (D and E). (F) Box plots showing [Ca 2+ ] i  peak response to cooling ramps (peak value of the F 340 /F 380  ratio during the ramp) in cold sensitive corneal neurons of control (n = 36; median = 0.40, range = 0.21-1.87) and tear-deficient animals (29; median = 0.55, range = 0.21-2.10). Horizontal line, median; bar, 25th and 75th interquartile range; whiskers, 10th and 90th percentiles. (G) Box plots showing [Ca 2+ ] i  threshold response to cooling ramps (temperature decrease required to evoke the [Ca 2+ ] i  response) in the same population of corneal neurons of control (n = 36; median = −16.06, range = −4.79 to −20.59) and tear-deficient animals (n = 29; median = −17.23, range = −5.94 to −23.46).
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    Thermo Fisher triethylammonium acetate solution
    Intracellular calcium responses of trigeminal sensory neurons innervating the cornea of control and 4-week tear-deficient guinea pigs. (A) Trigeminal ganglion (TG) neurons innervating the cornea were retrogradely labeled with FM 1-43 applied on the corneal surface 6 days earlier and identified with fluorescence microscopy. Only one or 2 corneal neurons were found per culture plate. Photographs show (from left to right) bright field, FM 1-43 fluorescence when excited with 470-nm light and merged images. (B–E) Ratiometric fluorescence changes (expressed as the ratio of fluorescence of Fura-2 when excited at 340 and 380 nm, F 340 /F 380 ) recorded simultaneously in the same culture plate from several fresh cultured TG sensory neurons from control (B and D) and tear-deficient (C and E) animals, in response to a cooling ramp and to the addition to the bath solution of 100 μM menthol, 100 μM cinnamaldehyde, 1 μM capsaicin, and 30 mM K + ; the bath temperature was simultaneously recorded (lower channels). The blue lines correspond to recordings from TG neurons innervating the cornea. Neurons responding to cooling and menthol were classified as cold thermosensitive neurons (B and C); neurons not responding to cooling stimulus but responding to capsaicin and cinnamaldehyde were classified as putative polymodal nociceptive neurons (D and E). (F) Box plots showing [Ca 2+ ] i  peak response to cooling ramps (peak value of the F 340 /F 380  ratio during the ramp) in cold sensitive corneal neurons of control (n = 36; median = 0.40, range = 0.21-1.87) and tear-deficient animals (29; median = 0.55, range = 0.21-2.10). Horizontal line, median; bar, 25th and 75th interquartile range; whiskers, 10th and 90th percentiles. (G) Box plots showing [Ca 2+ ] i  threshold response to cooling ramps (temperature decrease required to evoke the [Ca 2+ ] i  response) in the same population of corneal neurons of control (n = 36; median = −16.06, range = −4.79 to −20.59) and tear-deficient animals (n = 29; median = −17.23, range = −5.94 to −23.46).
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    Thermo Fisher quantification fm 1 43 labeling
    Photoinactivation of Dynamin results in the formation of large membrane inclusions.  (A–H) FM 1-43 labeling in  yw  and  shi 12-12B ;  shi-4C  animals treated (+) or not treated (−) with FlAsH for 10 min and/or illumination for 2 min (±). All preparations were stimulated with KCl in the presence of FM 1-43 for 5 min. (I) Quantification of the number of FM 1-43–labeled membrane accumulations (accum.) per boutonic area in  yw  controls ( n  = 72 boutons from eight larvae) and in  shi 12-12B ;  shi-4C  after FALI ( n  = 72 boutons from 16 larvae). Error bars show SEMs;  t  test: ***, P
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    Image Search Results


    Reaction scheme for creating click-conjugated dendrons employed in the in vitro studies. (i) glutaric anhydride, MeOH, 24 hrs, r.t. (ii) EDC, c(RGDyK), H 2 O/DMSO, 2 days, r.t. (iii) Alexa Fluor 488 azide OR biotin-dPEG 3+4 -azide OR MTX - azide (5) OP azide-G2(AF)

    Journal: Bioconjugate chemistry

    Article Title: RGD Dendron bodies; synthetic avidity agents with defined and potentially interchangeable effector sites that can substitute for antibodies

    doi: 10.1021/bc900217h

    Figure Lengend Snippet: Reaction scheme for creating click-conjugated dendrons employed in the in vitro studies. (i) glutaric anhydride, MeOH, 24 hrs, r.t. (ii) EDC, c(RGDyK), H 2 O/DMSO, 2 days, r.t. (iii) Alexa Fluor 488 azide OR biotin-dPEG 3+4 -azide OR MTX - azide (5) OP azide-G2(AF)

    Article Snippet: Alexa Fluor 488 azide and Prolong Gold mounting agent were obtained from Invitrogen. c(RGDyK) peptide was obtained from Peptide International.

    Techniques: In Vitro

    Dendron conjugate AF-G3(COOH) 11.9 (RGD) 4.1 (4) where Alexa Fluor 488 is conjugated to the dendron focal point via a 1,3-dipolar cycloaddition. The targeting moiety, c(RGDyK) peptide, is conjugated to the carboxylated surface of the dendron.

    Journal: Bioconjugate chemistry

    Article Title: RGD Dendron bodies; synthetic avidity agents with defined and potentially interchangeable effector sites that can substitute for antibodies

    doi: 10.1021/bc900217h

    Figure Lengend Snippet: Dendron conjugate AF-G3(COOH) 11.9 (RGD) 4.1 (4) where Alexa Fluor 488 is conjugated to the dendron focal point via a 1,3-dipolar cycloaddition. The targeting moiety, c(RGDyK) peptide, is conjugated to the carboxylated surface of the dendron.

    Article Snippet: Alexa Fluor 488 azide and Prolong Gold mounting agent were obtained from Invitrogen. c(RGDyK) peptide was obtained from Peptide International.

    Techniques:

    Analysis of domain-specific TRITC-DHPE diffusivity ( A ) and E raft data of NBD-DHPE distribution ( B ) suggest that variation in membrane CHOL between 25 and 37 mol% CHOL alters lipid packing differences between l o and l d domains of a polymer-tethered lipid bilayer containing DOPC/DPPC/CHOL mixtures (1:1 DOPC/DPPC molar ratio); p

    Journal: Biophysical Journal

    Article Title: Changes in Cholesterol Level Alter Integrin Sequestration in Raft-Mimicking Lipid Mixtures

    doi: 10.1016/j.bpj.2017.11.005

    Figure Lengend Snippet: Analysis of domain-specific TRITC-DHPE diffusivity ( A ) and E raft data of NBD-DHPE distribution ( B ) suggest that variation in membrane CHOL between 25 and 37 mol% CHOL alters lipid packing differences between l o and l d domains of a polymer-tethered lipid bilayer containing DOPC/DPPC/CHOL mixtures (1:1 DOPC/DPPC molar ratio); p

    Article Snippet: The dye-labeled phospholipids N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)-1,2-dihexadec-anoyl- sn -glycero-3-phosphoethanolamine, triethylammonium salt (NBD-DHPE) and N-(6-tetramethylrhodamine-thiocarbamoyl)-1, 2-dihexadecanayl- sn -glycero-3-phosphoethanolamine, triethylammonium salt (TRITC-DHPE) were obtained from Invitrogen (Carlsbad, CA).

    Techniques:

    Imaging of enhanced release with FM2-10 styryl dye. A , NMJs were first loaded with FM2-10 by stimulating 90 s at 10 Hz and then challenged (separately) with different stimulation trains. B , In response to 120 s at 50 Hz stimulation train, both WT and R6/1 synapses lost the FM2-10 dye almost completely. No obvious changes in destaining kinetics were observed between WT and R6/1. C , D , Upon milder stimulation with 40 s at 10 Hz trains, mutant synapses destained more and faster than WT synapses, as expected for higher release probability at R6/1 synapses. E , Recycling pool recovery from depletion (induced by a 30 s at 100 Hz train) was assessed by measuring EPP amplitude evoked by sequential stimulation at 2 Hz. Although the depression is stronger for R6/1 synapses (for clarity, only 1 point of 10 is drawn), the recovery was very similar in WT and mutant synapses. Thus, no obvious abnormalities occurred in the refilling of the recycling pool at R6/1 synapses.

    Journal: The Journal of Neuroscience

    Article Title: Increased Neurotransmitter Release at the Neuromuscular Junction in a Mouse Model of Polyglutamine Disease

    doi: 10.1523/JNEUROSCI.2011-10.2011

    Figure Lengend Snippet: Imaging of enhanced release with FM2-10 styryl dye. A , NMJs were first loaded with FM2-10 by stimulating 90 s at 10 Hz and then challenged (separately) with different stimulation trains. B , In response to 120 s at 50 Hz stimulation train, both WT and R6/1 synapses lost the FM2-10 dye almost completely. No obvious changes in destaining kinetics were observed between WT and R6/1. C , D , Upon milder stimulation with 40 s at 10 Hz trains, mutant synapses destained more and faster than WT synapses, as expected for higher release probability at R6/1 synapses. E , Recycling pool recovery from depletion (induced by a 30 s at 100 Hz train) was assessed by measuring EPP amplitude evoked by sequential stimulation at 2 Hz. Although the depression is stronger for R6/1 synapses (for clarity, only 1 point of 10 is drawn), the recovery was very similar in WT and mutant synapses. Thus, no obvious abnormalities occurred in the refilling of the recycling pool at R6/1 synapses.

    Article Snippet: For FM2-10 destaining experiments, LAL muscles were dissected and stretched in Ringer solution. d -Tubocurarine was used to avoid muscle contraction, and the muscle was incubated in FM2-10 (Invitrogen, 40 μ m ) for 5 min before the stimulation was applied (90 s at 10 Hz).

    Techniques: Imaging, Mutagenesis

    The status of the membranous organelles (MOs) in sperm. A) When labeled with the membrane dye FM® 1–43 (Life Technologies™), fused MOs are visible as bright spots just inside the cell membrane, but unfused MOs are not labeled. Here, the sperm are shown in both DIC illumination and epifluorescence illumination. Top panel: a spermatozoon with an obvious pseudopod and the bright foci in the cell body indicating fused MOs. Middle panel: an abnormal spermatid with fused MOs. Bottom panel: a normal spermatid with no fused MOs. B) The percentage of normal spermatids and those that had at least a single obvious fused MO from  him-5  and  spe-46; him-5  male worms. Error bars represent SEM. In C-F, the sperm have been fixed, permeabilized, and exposed to an Alexa Fluor® 594 conjugate of wheat germ agglutinin (WGA; Life Technologies™), which labels all MOs, including those that have not fused with the cell membrane (original red fluorescence false colored green). C) Sperm from a  spe-46(hc197); him-5(e1490)  male. D) The same cells as is C, visualized in DIC optics. E) Sperm from a  him-5(e1490)  male labeled as in C. F) DIC image of the cells in E.

    Journal: PLoS ONE

    Article Title: Premature Sperm Activation and Defective Spermatogenesis Caused by Loss of spe-46 Function in Caenorhabditis elegans

    doi: 10.1371/journal.pone.0057266

    Figure Lengend Snippet: The status of the membranous organelles (MOs) in sperm. A) When labeled with the membrane dye FM® 1–43 (Life Technologies™), fused MOs are visible as bright spots just inside the cell membrane, but unfused MOs are not labeled. Here, the sperm are shown in both DIC illumination and epifluorescence illumination. Top panel: a spermatozoon with an obvious pseudopod and the bright foci in the cell body indicating fused MOs. Middle panel: an abnormal spermatid with fused MOs. Bottom panel: a normal spermatid with no fused MOs. B) The percentage of normal spermatids and those that had at least a single obvious fused MO from him-5 and spe-46; him-5 male worms. Error bars represent SEM. In C-F, the sperm have been fixed, permeabilized, and exposed to an Alexa Fluor® 594 conjugate of wheat germ agglutinin (WGA; Life Technologies™), which labels all MOs, including those that have not fused with the cell membrane (original red fluorescence false colored green). C) Sperm from a spe-46(hc197); him-5(e1490) male. D) The same cells as is C, visualized in DIC optics. E) Sperm from a him-5(e1490) male labeled as in C. F) DIC image of the cells in E.

    Article Snippet: The lipid dye FM®1–43 (Life Technologies™) labels the cell membrane, and if any MOs have fused, they become labeled by FM®1–43 as well resulting in fluorescent foci abutting the cell membrane .

    Techniques: Labeling, Whole Genome Amplification, Fluorescence

    The PLD antagonist  n -ButOH and DGK inhibitor R59022 disrupt internalization of the endomembrane tracer dye FM 1-43 .  (A)  Ninety-minute-old pollen tubes were pre-incubated in germination medium on ice for 10 min, and 2 μM FM1-43 dye together with a particular drug or PA were then simultaneously added and the cells monitored for 30 min. Representative pollen tubes observed at two times are shown.  (B)  Alternatively, pollen tubes were incubated in germination medium containing 2 μM FM1-43 for 90 min, a particular drug or PA were then added for 10 min and the cells were observed with a confocal microscope. At least 15 cells were analyzed for each treatment in four independent experiments and typical examples are shown. Scale bar = 10 μm.

    Journal: Frontiers in plant science

    Article Title: Turnover of Phosphatidic Acid through Distinct Signaling Pathways Affects Multiple Aspects of Pollen Tube Growth in Tobacco

    doi: 10.3389/fpls.2012.00054

    Figure Lengend Snippet: The PLD antagonist n -ButOH and DGK inhibitor R59022 disrupt internalization of the endomembrane tracer dye FM 1-43 . (A) Ninety-minute-old pollen tubes were pre-incubated in germination medium on ice for 10 min, and 2 μM FM1-43 dye together with a particular drug or PA were then simultaneously added and the cells monitored for 30 min. Representative pollen tubes observed at two times are shown. (B) Alternatively, pollen tubes were incubated in germination medium containing 2 μM FM1-43 for 90 min, a particular drug or PA were then added for 10 min and the cells were observed with a confocal microscope. At least 15 cells were analyzed for each treatment in four independent experiments and typical examples are shown. Scale bar = 10 μm.

    Article Snippet: FM 1-43 labeling, vacuole, pectin and callose staining, actin labeling Tobacco pollen tubes were labeled with endocytic marker FM 1-43 (Molecular Probes).

    Techniques: Incubation, Microscopy

    CRISPRi-mediated phenotypes associated with biofilm structure and EPS matrix. P . fluorescens SBW25 cells expressing the CRISPRi system targeting genes gacS (PFLU3777, two-component sensor kinase), rimA (PFLU0263, PDE), dipA (PFLU0458, PDE), alg44 (PFLU0988, alginase co-polymerase Alg44), gcbA (PFL0621, DGC) and bifA (PFLU4858, PDE). The control corresponds to the pPFL-gRNA plasmid with no guide RNA inserted. (A) Cells were stained with FM 1–43 and observed by confocal microscopy. 3D architecture of biofilms were reconstructed as described in Methods. Virtual shadow projections were included on the right to show thickness. (B) Biofilms grown in the presence of the Congo Red dye to reveal the structure of EPS. The ESP-stained volumes were rendered with maximum intensity projection. Inset shows a blow-up view of a clump in the EPS matrix after silencing of bifA .

    Journal: Scientific Reports

    Article Title: CRISPR interference to interrogate genes that control biofilm formation in Pseudomonas fluorescens

    doi: 10.1038/s41598-019-52400-5

    Figure Lengend Snippet: CRISPRi-mediated phenotypes associated with biofilm structure and EPS matrix. P . fluorescens SBW25 cells expressing the CRISPRi system targeting genes gacS (PFLU3777, two-component sensor kinase), rimA (PFLU0263, PDE), dipA (PFLU0458, PDE), alg44 (PFLU0988, alginase co-polymerase Alg44), gcbA (PFL0621, DGC) and bifA (PFLU4858, PDE). The control corresponds to the pPFL-gRNA plasmid with no guide RNA inserted. (A) Cells were stained with FM 1–43 and observed by confocal microscopy. 3D architecture of biofilms were reconstructed as described in Methods. Virtual shadow projections were included on the right to show thickness. (B) Biofilms grown in the presence of the Congo Red dye to reveal the structure of EPS. The ESP-stained volumes were rendered with maximum intensity projection. Inset shows a blow-up view of a clump in the EPS matrix after silencing of bifA .

    Article Snippet: The cover slides with intact biofilm pellicles were then mounted on an Attofluor™ Cell Chamber, covered with 1 ml of PBS or transparent minimal media and stained with the FilmTracer FM 1–43 Green Biofilm dye (Molecular Probe) for 30 min prior to observation by confocal microscopy.

    Techniques: Expressing, Plasmid Preparation, Staining, Confocal Microscopy, End-sequence Profiling

    Bilayer calibration curves and calculation of binding ratio. ( A ) Intensity of NBD-PC bilayer at different molecular densities with the F factor comparing brightness of Notch1-eGFP and NBD-PC. ( B ) Calibration curve and F factor relating TRITC-DHPE and

    Journal: Biophysical Journal

    Article Title: Membrane Tethered Delta Activates Notch and Reveals a Role for Spatio-Mechanical Regulation of the Signaling Pathway

    doi: 10.1016/j.bpj.2013.11.012

    Figure Lengend Snippet: Bilayer calibration curves and calculation of binding ratio. ( A ) Intensity of NBD-PC bilayer at different molecular densities with the F factor comparing brightness of Notch1-eGFP and NBD-PC. ( B ) Calibration curve and F factor relating TRITC-DHPE and

    Article Snippet: Fluorescent lipid ( N -(6-tetramethylrhodaminethiocarbamoyl)-1,2-dihexadecanoyl- sn -glycero-3-phosphoethanolamine), TRITC-DHPE was from Life Technologies (Grand Island, NY).

    Techniques: Binding Assay

    Generation of antioxidant over-expressing parasites. Transgenic parasites were created in the RHΔhxgprt background strain using plasmids designed to over-express HA-tagged versions of glutaredoxin (Glut), catalase (Cat), or peroxiredoxin 2 (PRx2) under the control of the gra1 gene promoter. ( a ) Over-expression was verified using quantitative real-time PCR to measure transcript levels. ( b ) Western blot analysis was used to confirm protein production and that the resultant proteins were the appropriate size. ( c ) Immunofluorescence assays were performed for protein localization, utilizing a rabbit monoclonal antibody against the HA tag and an Alexa Fluor 594 secondary antibody. ( d ) General parasite replication was determined by doubling assay. Confluent HFF in 24-well plates were infected with the indicated parasite lines for 5 hours before the medium was replaced with fresh growth medium. Parasites were allowed to grow for 32 hours before being methanol-fixed and stained with DiffQuick. The number of individual parasites per vacuole was enumerated for at least 50 vacuoles. Data is presented as percent of vacuoles from 4 experiments. Student’s t-test was employed for statistical analysis. Scale bars equal 2 μm.

    Journal: Scientific Reports

    Article Title: Oxidative stress generated during monensin treatment contributes to altered Toxoplasma gondii mitochondrial function

    doi: 10.1038/srep22997

    Figure Lengend Snippet: Generation of antioxidant over-expressing parasites. Transgenic parasites were created in the RHΔhxgprt background strain using plasmids designed to over-express HA-tagged versions of glutaredoxin (Glut), catalase (Cat), or peroxiredoxin 2 (PRx2) under the control of the gra1 gene promoter. ( a ) Over-expression was verified using quantitative real-time PCR to measure transcript levels. ( b ) Western blot analysis was used to confirm protein production and that the resultant proteins were the appropriate size. ( c ) Immunofluorescence assays were performed for protein localization, utilizing a rabbit monoclonal antibody against the HA tag and an Alexa Fluor 594 secondary antibody. ( d ) General parasite replication was determined by doubling assay. Confluent HFF in 24-well plates were infected with the indicated parasite lines for 5 hours before the medium was replaced with fresh growth medium. Parasites were allowed to grow for 32 hours before being methanol-fixed and stained with DiffQuick. The number of individual parasites per vacuole was enumerated for at least 50 vacuoles. Data is presented as percent of vacuoles from 4 experiments. Student’s t-test was employed for statistical analysis. Scale bars equal 2 μm.

    Article Snippet: Primary antibodies were visualized using either Alexa Fluor 594 or Alexa Fluor 488 secondary antibodies (Life Technologies).

    Techniques: Expressing, Transgenic Assay, Over Expression, Real-time Polymerase Chain Reaction, Western Blot, Immunofluorescence, Infection, Staining

    Images of immunofluorescence by confocal laser scanning microscopy (CLSM) of four different breast cancer cell lines: MCF-7, SKBR3, T47D and MDA-MB-231 ( a ) bright field; ( b ) staining of the cell nucleus by the fluorophore DAPI (blue); ( c ) indirect detection of HER2 by a primary anti-HER2 antibody and an anti-anti-HER2 secondary antibody bound to Alexa Fluor 594 fluorophore (red) and ( d ) cell localization of HER2 receptor by overlapping of images (DAPI + HER2). Magnification: 63×.

    Journal: Nanomaterials

    Article Title: Development of a Nanostructured Platform for Identifying HER2-Heterogeneity of Breast Cancer Cells by Surface-Enhanced Raman Scattering

    doi: 10.3390/nano8070549

    Figure Lengend Snippet: Images of immunofluorescence by confocal laser scanning microscopy (CLSM) of four different breast cancer cell lines: MCF-7, SKBR3, T47D and MDA-MB-231 ( a ) bright field; ( b ) staining of the cell nucleus by the fluorophore DAPI (blue); ( c ) indirect detection of HER2 by a primary anti-HER2 antibody and an anti-anti-HER2 secondary antibody bound to Alexa Fluor 594 fluorophore (red) and ( d ) cell localization of HER2 receptor by overlapping of images (DAPI + HER2). Magnification: 63×.

    Article Snippet: The following materials were used: Dulbecco’s modified eagle’s medium (DMEM), fetal bovine serum (FBS), RPMI medium, F12 medium, non-essential amino acids, l -glutamine, sodium pyruvate, nunclon 6-well multidish, Alexa Fluor 594 dye (secondary antibody) and Penicillin/Streptomycin (antibiotic/antifungal) were purchased from Thermo Fisher Scientific, Mexico City, Mexico.

    Techniques: Immunofluorescence, Confocal Laser Scanning Microscopy, Multiple Displacement Amplification, Staining

    Time-lapse fluorescence microscopy images of shrinking vesicles. (a) The GFP leaked from DOPC/ 5 GUV upon the shrinkage of the vesicle by PLA 2 treatment. The GUV membrane was stained with Texas Red DHPE. (b) The 100 nm red fluorescent sulfate-modified polystyrene nanoparticles were contained and concentrated in DOPC/ 5 GUV as the vesicle shrank. The membrane was stained with NBD-PE.

    Journal: Chemical Science

    Article Title: Rapid access to phospholipid analogs using thiol-yne chemistry access to phospholipid analogs using thiol-yne chemistry †Electronic supplementary information (ESI) available: Details of experimental procedures, lipid characterization data (NMR, HRMS, steady-state fluorescence anisotropy), HPLC analysis, fluorescence microscopy images, liposome shrinkage data in the form of AVI files. See DOI: 10.1039/c5sc00653hClick here for additional data file.

    doi: 10.1039/c5sc00653h

    Figure Lengend Snippet: Time-lapse fluorescence microscopy images of shrinking vesicles. (a) The GFP leaked from DOPC/ 5 GUV upon the shrinkage of the vesicle by PLA 2 treatment. The GUV membrane was stained with Texas Red DHPE. (b) The 100 nm red fluorescent sulfate-modified polystyrene nanoparticles were contained and concentrated in DOPC/ 5 GUV as the vesicle shrank. The membrane was stained with NBD-PE.

    Article Snippet: 0.5 μL of 100 mM Texas Red DHPE or NBD-PE (Invitrogen, Carlsbad, CA) was added to 100 μL vesicle suspension, which was then added to a 96-well plate.

    Techniques: Fluorescence, Microscopy, Proximity Ligation Assay, Staining, Modification

    (a) Flow analysis to show aggregated human γ-globulin (AHG)–AlexaFluor® 488 binding to CD4 +  T cells in purified peripheral blood mononuclear cells (PBMCs). Cells were gated using forward- and side-scatter. CD4 +  lymphocytes were

    Journal: Clinical and Experimental Immunology

    Article Title: Immune complexes and late complement proteins trigger activation of Syk tyrosine kinase in human CD4+ T cells

    doi: 10.1111/j.1365-2249.2011.04505.x

    Figure Lengend Snippet: (a) Flow analysis to show aggregated human γ-globulin (AHG)–AlexaFluor® 488 binding to CD4 + T cells in purified peripheral blood mononuclear cells (PBMCs). Cells were gated using forward- and side-scatter. CD4 + lymphocytes were

    Article Snippet: One mg of the AHG protein was labelled with AlexaFluor® 488 using the protein labelling kit, as per the manufacturer's protocol (Invitrogen).

    Techniques: Flow Cytometry, Binding Assay, Purification

    Comparison of OATP1B1- and OATP1B3-mediated transport of fluorescent substrates. Uptake was measured with 1 µM of the indicated fluorescent compounds on 24-well plates for 30 min. Values are means ± SE of two combined independent experiments each performed with triplicate determinations. FMTX: fluorescein methotrexate; AMTX: Alexa Fluor® 488 methotrexate; Flutax-2: Oregon Green® 488 Taxol; WT: wild-type CHO cells; OATP1B1: CHO cells expressing OATP1B1; OATP1B3: CHO cells expressing OATP1B3.

    Journal: Current Chemical Genomics

    Article Title: Development of a Cell-Based High-Throughput Assay to Screen for Inhibitors of Organic Anion Transporting Polypeptides 1B1 and 1B3

    doi: 10.2174/1875397301004010001

    Figure Lengend Snippet: Comparison of OATP1B1- and OATP1B3-mediated transport of fluorescent substrates. Uptake was measured with 1 µM of the indicated fluorescent compounds on 24-well plates for 30 min. Values are means ± SE of two combined independent experiments each performed with triplicate determinations. FMTX: fluorescein methotrexate; AMTX: Alexa Fluor® 488 methotrexate; Flutax-2: Oregon Green® 488 Taxol; WT: wild-type CHO cells; OATP1B1: CHO cells expressing OATP1B1; OATP1B3: CHO cells expressing OATP1B3.

    Article Snippet: Materials Cell culture reagents, fluorescein-methotrexate (FMTX), Alexa Fluor® 488 methotrexate (AMTX), and Oregon Green® 488 Taxol (Flutax-2) were purchased from Invitrogen (Carlsbad, CA).

    Techniques: Expressing

    Characterization of biogenic Mn oxides produced by strain T-G1 in PYG medium at pH 5.5. (A)  Scanning electron micrograph showing spherical clusters of micron to sub-micron sheets of Mn oxide minerals.  (B)  An enlargement of the area indicated by a red box in  (A) .  (C)  Confocal laser scanning microscopy (CLSM) of a mineral aggregate in association with T-G1. Bacterial cells stained by FM 1-43 probe shown in green, reflection of minerals was indicated in gray.  (D)  Energy-dispersive X-ray (EDS) spectra showing elemental composition of biogenic Mn oxide particles.  (E)  Raman spectroscopy of biogenic Mn oxides.  (F)  XRD pattern of biogenic Mn-oxide minerals with vertical lines indicating strong (red) and weak peak positions (green) of bixbyite.

    Journal: Frontiers in Microbiology

    Article Title: Characterization of pH dependent Mn(II) oxidation strategies and formation of a bixbyite-like phase by Mesorhizobium australicum T-G1

    doi: 10.3389/fmicb.2015.00734

    Figure Lengend Snippet: Characterization of biogenic Mn oxides produced by strain T-G1 in PYG medium at pH 5.5. (A) Scanning electron micrograph showing spherical clusters of micron to sub-micron sheets of Mn oxide minerals. (B) An enlargement of the area indicated by a red box in (A) . (C) Confocal laser scanning microscopy (CLSM) of a mineral aggregate in association with T-G1. Bacterial cells stained by FM 1-43 probe shown in green, reflection of minerals was indicated in gray. (D) Energy-dispersive X-ray (EDS) spectra showing elemental composition of biogenic Mn oxide particles. (E) Raman spectroscopy of biogenic Mn oxides. (F) XRD pattern of biogenic Mn-oxide minerals with vertical lines indicating strong (red) and weak peak positions (green) of bixbyite.

    Article Snippet: Bacterial cells were stained using the nucleic acid stain Syto 9 and the membrane stain FM 1-43 (Molecular Probes, Eugene, OR, USA).

    Techniques: Produced, Confocal Laser Scanning Microscopy, Staining, Raman Spectroscopy

    Intracellular calcium responses of trigeminal sensory neurons innervating the cornea of control and 4-week tear-deficient guinea pigs. (A) Trigeminal ganglion (TG) neurons innervating the cornea were retrogradely labeled with FM 1-43 applied on the corneal surface 6 days earlier and identified with fluorescence microscopy. Only one or 2 corneal neurons were found per culture plate. Photographs show (from left to right) bright field, FM 1-43 fluorescence when excited with 470-nm light and merged images. (B–E) Ratiometric fluorescence changes (expressed as the ratio of fluorescence of Fura-2 when excited at 340 and 380 nm, F 340 /F 380 ) recorded simultaneously in the same culture plate from several fresh cultured TG sensory neurons from control (B and D) and tear-deficient (C and E) animals, in response to a cooling ramp and to the addition to the bath solution of 100 μM menthol, 100 μM cinnamaldehyde, 1 μM capsaicin, and 30 mM K + ; the bath temperature was simultaneously recorded (lower channels). The blue lines correspond to recordings from TG neurons innervating the cornea. Neurons responding to cooling and menthol were classified as cold thermosensitive neurons (B and C); neurons not responding to cooling stimulus but responding to capsaicin and cinnamaldehyde were classified as putative polymodal nociceptive neurons (D and E). (F) Box plots showing [Ca 2+ ] i  peak response to cooling ramps (peak value of the F 340 /F 380  ratio during the ramp) in cold sensitive corneal neurons of control (n = 36; median = 0.40, range = 0.21-1.87) and tear-deficient animals (29; median = 0.55, range = 0.21-2.10). Horizontal line, median; bar, 25th and 75th interquartile range; whiskers, 10th and 90th percentiles. (G) Box plots showing [Ca 2+ ] i  threshold response to cooling ramps (temperature decrease required to evoke the [Ca 2+ ] i  response) in the same population of corneal neurons of control (n = 36; median = −16.06, range = −4.79 to −20.59) and tear-deficient animals (n = 29; median = −17.23, range = −5.94 to −23.46).

    Journal: Pain

    Article Title: Abnormal activity of corneal cold thermoreceptors underlies the unpleasant sensations in dry eye disease

    doi: 10.1097/j.pain.0000000000000455

    Figure Lengend Snippet: Intracellular calcium responses of trigeminal sensory neurons innervating the cornea of control and 4-week tear-deficient guinea pigs. (A) Trigeminal ganglion (TG) neurons innervating the cornea were retrogradely labeled with FM 1-43 applied on the corneal surface 6 days earlier and identified with fluorescence microscopy. Only one or 2 corneal neurons were found per culture plate. Photographs show (from left to right) bright field, FM 1-43 fluorescence when excited with 470-nm light and merged images. (B–E) Ratiometric fluorescence changes (expressed as the ratio of fluorescence of Fura-2 when excited at 340 and 380 nm, F 340 /F 380 ) recorded simultaneously in the same culture plate from several fresh cultured TG sensory neurons from control (B and D) and tear-deficient (C and E) animals, in response to a cooling ramp and to the addition to the bath solution of 100 μM menthol, 100 μM cinnamaldehyde, 1 μM capsaicin, and 30 mM K + ; the bath temperature was simultaneously recorded (lower channels). The blue lines correspond to recordings from TG neurons innervating the cornea. Neurons responding to cooling and menthol were classified as cold thermosensitive neurons (B and C); neurons not responding to cooling stimulus but responding to capsaicin and cinnamaldehyde were classified as putative polymodal nociceptive neurons (D and E). (F) Box plots showing [Ca 2+ ] i peak response to cooling ramps (peak value of the F 340 /F 380 ratio during the ramp) in cold sensitive corneal neurons of control (n = 36; median = 0.40, range = 0.21-1.87) and tear-deficient animals (29; median = 0.55, range = 0.21-2.10). Horizontal line, median; bar, 25th and 75th interquartile range; whiskers, 10th and 90th percentiles. (G) Box plots showing [Ca 2+ ] i threshold response to cooling ramps (temperature decrease required to evoke the [Ca 2+ ] i response) in the same population of corneal neurons of control (n = 36; median = −16.06, range = −4.79 to −20.59) and tear-deficient animals (n = 29; median = −17.23, range = −5.94 to −23.46).

    Article Snippet: Labelling of trigeminal ganglion corneal neurons In guinea pigs anesthetized with 90 mg/kg ketamine and 5 mg/kg xylazine (i.p.), a 6 mm-diameter piece of Spongostan film (Ferrosan A/S, Soeborg, Denmark) saturated with the cell stain FM 1-43 (Molecular probes; Invitrogen) at 5 mM in saline solution was carefully placed in the center of the cornea to retrogradely label TG sensory cell bodies whose axons innervate the cornea.

    Techniques: Labeling, Fluorescence, Microscopy, Cell Culture

    Voltage-gated Na +  currents in trigeminal polymodal sensory neurons of control and tear-deficient guinea pigs 4 weeks after lachrymal gland removal. Corneal neurons were retrogradely labeled with FM 1-43 applied on the cornea 6 days earlier. Mean voltage–current relationships for TTX-r (A) and TTX-s (B) sodium currents. Data are mean ± SEM; ** P

    Journal: Pain

    Article Title: Abnormal activity of corneal cold thermoreceptors underlies the unpleasant sensations in dry eye disease

    doi: 10.1097/j.pain.0000000000000455

    Figure Lengend Snippet: Voltage-gated Na + currents in trigeminal polymodal sensory neurons of control and tear-deficient guinea pigs 4 weeks after lachrymal gland removal. Corneal neurons were retrogradely labeled with FM 1-43 applied on the cornea 6 days earlier. Mean voltage–current relationships for TTX-r (A) and TTX-s (B) sodium currents. Data are mean ± SEM; ** P

    Article Snippet: Labelling of trigeminal ganglion corneal neurons In guinea pigs anesthetized with 90 mg/kg ketamine and 5 mg/kg xylazine (i.p.), a 6 mm-diameter piece of Spongostan film (Ferrosan A/S, Soeborg, Denmark) saturated with the cell stain FM 1-43 (Molecular probes; Invitrogen) at 5 mM in saline solution was carefully placed in the center of the cornea to retrogradely label TG sensory cell bodies whose axons innervate the cornea.

    Techniques: Labeling

    Voltage-gated Na +  currents in trigeminal corneal cold sensory neurons of control and tear-deficient guinea pigs at 4 weeks after lachrymal gland removal. Corneal neurons were retrogradely labeled with FM 1-43 applied on the cornea 6 days earlier. (A) For each neuron, membrane potential was held at −80 mV; whole-cell sodium currents were evoked after a 500-millisecond prepulse to either −120 or −40 mV with a 100-millisecond step to potentials between −60 and +50 mV in 10 mV increments; current evoked from −40 mV was considered to be TTX-r current, whereas the difference between the current evoked from −120 and −40 mV was considered to be TTX-s; for clarity, only traces from −60 to +20 mV are shown. (B and C) Mean voltage–current relationships and mean relative peak conductance normalized to the maximal conductance (G/G max ) and plotted against voltage of TTX-r sodium currents. (D and E) Mean voltage–current relationships and mean relative peak conductance normalized to the maximal conductance (G/G max ) and plotted against voltage of TTX-s sodium currents. Data are mean ± SEM; ** P

    Journal: Pain

    Article Title: Abnormal activity of corneal cold thermoreceptors underlies the unpleasant sensations in dry eye disease

    doi: 10.1097/j.pain.0000000000000455

    Figure Lengend Snippet: Voltage-gated Na + currents in trigeminal corneal cold sensory neurons of control and tear-deficient guinea pigs at 4 weeks after lachrymal gland removal. Corneal neurons were retrogradely labeled with FM 1-43 applied on the cornea 6 days earlier. (A) For each neuron, membrane potential was held at −80 mV; whole-cell sodium currents were evoked after a 500-millisecond prepulse to either −120 or −40 mV with a 100-millisecond step to potentials between −60 and +50 mV in 10 mV increments; current evoked from −40 mV was considered to be TTX-r current, whereas the difference between the current evoked from −120 and −40 mV was considered to be TTX-s; for clarity, only traces from −60 to +20 mV are shown. (B and C) Mean voltage–current relationships and mean relative peak conductance normalized to the maximal conductance (G/G max ) and plotted against voltage of TTX-r sodium currents. (D and E) Mean voltage–current relationships and mean relative peak conductance normalized to the maximal conductance (G/G max ) and plotted against voltage of TTX-s sodium currents. Data are mean ± SEM; ** P

    Article Snippet: Labelling of trigeminal ganglion corneal neurons In guinea pigs anesthetized with 90 mg/kg ketamine and 5 mg/kg xylazine (i.p.), a 6 mm-diameter piece of Spongostan film (Ferrosan A/S, Soeborg, Denmark) saturated with the cell stain FM 1-43 (Molecular probes; Invitrogen) at 5 mM in saline solution was carefully placed in the center of the cornea to retrogradely label TG sensory cell bodies whose axons innervate the cornea.

    Techniques: Labeling

    Photoinactivation of Dynamin results in the formation of large membrane inclusions.  (A–H) FM 1-43 labeling in  yw  and  shi 12-12B ;  shi-4C  animals treated (+) or not treated (−) with FlAsH for 10 min and/or illumination for 2 min (±). All preparations were stimulated with KCl in the presence of FM 1-43 for 5 min. (I) Quantification of the number of FM 1-43–labeled membrane accumulations (accum.) per boutonic area in  yw  controls ( n  = 72 boutons from eight larvae) and in  shi 12-12B ;  shi-4C  after FALI ( n  = 72 boutons from 16 larvae). Error bars show SEMs;  t  test: ***, P

    Journal: The Journal of Cell Biology

    Article Title: Dynamin photoinactivation blocks Clathrin and α-adaptin recruitment and induces bulk membrane retrieval

    doi: 10.1083/jcb.201310090

    Figure Lengend Snippet: Photoinactivation of Dynamin results in the formation of large membrane inclusions. (A–H) FM 1-43 labeling in yw and shi 12-12B ; shi-4C animals treated (+) or not treated (−) with FlAsH for 10 min and/or illumination for 2 min (±). All preparations were stimulated with KCl in the presence of FM 1-43 for 5 min. (I) Quantification of the number of FM 1-43–labeled membrane accumulations (accum.) per boutonic area in yw controls ( n = 72 boutons from eight larvae) and in shi 12-12B ; shi-4C after FALI ( n = 72 boutons from 16 larvae). Error bars show SEMs; t test: ***, P

    Article Snippet: Fluorescence imaging and quantification FM 1-43 labeling was performed by incubating dissected larvae either in HL-3 with 4 µM FM 1-43 (Invitrogen), 1.5 mM CaCl2 , and 90 mM KCl for 5 min (1 min for EndoA) or in HL-3 with 1.5 mM CaCl2 and electrically stimulating motor nerves at 10 Hz for 5 min.

    Techniques: Labeling

    Photoinactivation of Dynamin converts invaginated pits in  shi ts1  mutants into bulk cisternae.  (A and B) Quantification of the HA-Chc (A,  n  = 11 boutons from seven larvae each) and α-Ada (B,  n  = 7–10 boutons from six to seven larvae) labeling intensity in a line over the (largest) bouton diameter (and normalized to the length of the bouton diameter; see Materials and methods) in KCl-stimulated  shi ts1 /Y ;  HA-chc/+  at permissive (RT) temperature and  shi ts1 /Y ;  HA-chc/+  at restrictive temperature (33°C); see Materials and methods. Averages are in black lines, and SEMs are in gray shades. rel., relative. (C) Model of the distribution of different Dynamin molecules (Shi-4C and Shi ts1 ) in endocytic pits (based on anti-Dynamin labeling intensity—see Stimulus-dependent Clathrin and α-Ada recruitment in shi ts1  mutants in the Results section) and calculation of the probability that Dynamin rings consist only of Shi ts1  or only of Shi-4C in  shi ts1 /Y ;  shi-4C /+ larvae. (D–I) FM 1-43 labeling in  shi ts1 /Y ;  shi-4C/+  larvae treated (+; F–I) or not treated (−; D and E) with FlAsH for 10 min and illuminated for 2 min (+; G and I) at permissive (D, F, and G) or restrictive (E, H, and I) temperature. All preparations were stimulated for 5 min with KCl in the presence of FM 1-43. Note that  shi ts1 /Y ;  shi-4C/+  animals at restrictive temperature without FALI phenocopy  shi ts1  animals and do not internalize FM 1-43, whereas  shi ts1 /Y ;  shi-4C/+  animals at restrictive temperature after FALI also internalize FM 1-43 in cisternal inclusions, similar to  shi 12-12B ;  shi-4C  animals in which Dynamin was photoinactivated. Bar, 5 µm.

    Journal: The Journal of Cell Biology

    Article Title: Dynamin photoinactivation blocks Clathrin and α-adaptin recruitment and induces bulk membrane retrieval

    doi: 10.1083/jcb.201310090

    Figure Lengend Snippet: Photoinactivation of Dynamin converts invaginated pits in shi ts1 mutants into bulk cisternae. (A and B) Quantification of the HA-Chc (A, n = 11 boutons from seven larvae each) and α-Ada (B, n = 7–10 boutons from six to seven larvae) labeling intensity in a line over the (largest) bouton diameter (and normalized to the length of the bouton diameter; see Materials and methods) in KCl-stimulated shi ts1 /Y ; HA-chc/+ at permissive (RT) temperature and shi ts1 /Y ; HA-chc/+ at restrictive temperature (33°C); see Materials and methods. Averages are in black lines, and SEMs are in gray shades. rel., relative. (C) Model of the distribution of different Dynamin molecules (Shi-4C and Shi ts1 ) in endocytic pits (based on anti-Dynamin labeling intensity—see Stimulus-dependent Clathrin and α-Ada recruitment in shi ts1 mutants in the Results section) and calculation of the probability that Dynamin rings consist only of Shi ts1 or only of Shi-4C in shi ts1 /Y ; shi-4C /+ larvae. (D–I) FM 1-43 labeling in shi ts1 /Y ; shi-4C/+ larvae treated (+; F–I) or not treated (−; D and E) with FlAsH for 10 min and illuminated for 2 min (+; G and I) at permissive (D, F, and G) or restrictive (E, H, and I) temperature. All preparations were stimulated for 5 min with KCl in the presence of FM 1-43. Note that shi ts1 /Y ; shi-4C/+ animals at restrictive temperature without FALI phenocopy shi ts1 animals and do not internalize FM 1-43, whereas shi ts1 /Y ; shi-4C/+ animals at restrictive temperature after FALI also internalize FM 1-43 in cisternal inclusions, similar to shi 12-12B ; shi-4C animals in which Dynamin was photoinactivated. Bar, 5 µm.

    Article Snippet: Fluorescence imaging and quantification FM 1-43 labeling was performed by incubating dissected larvae either in HL-3 with 4 µM FM 1-43 (Invitrogen), 1.5 mM CaCl2 , and 90 mM KCl for 5 min (1 min for EndoA) or in HL-3 with 1.5 mM CaCl2 and electrically stimulating motor nerves at 10 Hz for 5 min.

    Techniques: Labeling