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manual tirf illuminator  (Nikon)
99
Nikon manual tirf illuminator
a Principles of STAR microscopy. The evanescent field decay is wavelength dependent, and STAR is based on the changing fluorescence ratio of two spectrally separated fluorophores. STAR imaging of CLCa dual-tagged with iRFP713 and EGFP can be used to study the curvature of clathrin-coated structures. b Current models describing formation of clathrin-coated vesicles (CCVs): i, Constant Curvature Model (CCM); ii, Flat-to-curved transition (FTC); iii, Flat clathrin lattice (FCL) accumulation and dispersion. c 5 μm silica microsphere dual-tagged with Alexa488 and Alexa647, imaged in epifluorescence (E) and <t>TIRF</t> (T), T647/T488 ratio, and theoretical (Theor—blue) and measured (Meas—red) z-position as a function of distance from the center of the bead, mean ± SEM from 31 beads. d Cos-7 cell expressing CLCa-iRFP713-EGFP (STAR) probe imaged simultaneously with 488 and 647 TIRF excitation, arrows indicate example clathrin accumulations, scale bar = 20 μm. Representative of 13 cells and five independent repeats. e Western-blot analysis of STAR probe overexpression in Cos-7 cells. Representative blot from two independent replicates. f Plot of transferrin internalization over time for WT (black) and STAR probe expressing (magenta) Cos-7 cells. Circles and triangles indicate means from three independent replicates at 0, 5, 10, 15, and 20 min for WT Cos-7 ( n = 93, 93, 93, 93, 93 cells) and CLCa-STAR OE ( n = 89, 91, 93, 86, 92 cells). Black and magenta lines represent means ± SD. There was no main effect of STAR probe expression on TF-568 uptake by two-way ANOVA [Probe, Stimulation]—[ F (1, 4) = 0.7909, p = 0.42; F (2.132, 8.526) = 189.5, p < 0.0001]. Šídák’s multiple comparisons test, details in Supplementary Table . g Kymograph showing of clathrin accumulation (cyan) and curvature formation (fire), full arrow indicates vesicle formation, dashed arrow indicates flat clathrin lattice assembly. The curvature channel is translated 3 pixels down for better visualization, scale bar = 5 μm. h Quantitative GFP (cyan) and iRFP713 (magenta) intensity and ∆z (orange) traces for representative CCV and FCL, darker shaded band—SD of detected signal. Lighter cyan, magenta, and orange indicate background mean and 2*SD above background, red dashed boxes indicate when GFP signal is significantly higher than threshold. a , b were created with BioRender.com.
Manual Tirf Illuminator, supplied by Nikon, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/manual+tirf+illuminator+unit/Objectives/pmc08976038-222-32-35
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manual nikon tirf illumination module  (Nikon)
95
Nikon manual nikon tirf illumination module
a Principles of STAR microscopy. The evanescent field decay is wavelength dependent, and STAR is based on the changing fluorescence ratio of two spectrally separated fluorophores. STAR imaging of CLCa dual-tagged with iRFP713 and EGFP can be used to study the curvature of clathrin-coated structures. b Current models describing formation of clathrin-coated vesicles (CCVs): i, Constant Curvature Model (CCM); ii, Flat-to-curved transition (FTC); iii, Flat clathrin lattice (FCL) accumulation and dispersion. c 5 μm silica microsphere dual-tagged with Alexa488 and Alexa647, imaged in epifluorescence (E) and <t>TIRF</t> (T), T647/T488 ratio, and theoretical (Theor—blue) and measured (Meas—red) z-position as a function of distance from the center of the bead, mean ± SEM from 31 beads. d Cos-7 cell expressing CLCa-iRFP713-EGFP (STAR) probe imaged simultaneously with 488 and 647 TIRF excitation, arrows indicate example clathrin accumulations, scale bar = 20 μm. Representative of 13 cells and five independent repeats. e Western-blot analysis of STAR probe overexpression in Cos-7 cells. Representative blot from two independent replicates. f Plot of transferrin internalization over time for WT (black) and STAR probe expressing (magenta) Cos-7 cells. Circles and triangles indicate means from three independent replicates at 0, 5, 10, 15, and 20 min for WT Cos-7 ( n = 93, 93, 93, 93, 93 cells) and CLCa-STAR OE ( n = 89, 91, 93, 86, 92 cells). Black and magenta lines represent means ± SD. There was no main effect of STAR probe expression on TF-568 uptake by two-way ANOVA [Probe, Stimulation]—[ F (1, 4) = 0.7909, p = 0.42; F (2.132, 8.526) = 189.5, p < 0.0001]. Šídák’s multiple comparisons test, details in Supplementary Table . g Kymograph showing of clathrin accumulation (cyan) and curvature formation (fire), full arrow indicates vesicle formation, dashed arrow indicates flat clathrin lattice assembly. The curvature channel is translated 3 pixels down for better visualization, scale bar = 5 μm. h Quantitative GFP (cyan) and iRFP713 (magenta) intensity and ∆z (orange) traces for representative CCV and FCL, darker shaded band—SD of detected signal. Lighter cyan, magenta, and orange indicate background mean and 2*SD above background, red dashed boxes indicate when GFP signal is significantly higher than threshold. a , b were created with BioRender.com.
Manual Nikon Tirf Illumination Module, supplied by Nikon, used in various techniques. Bioz Stars score: 95/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/manual+tirf+illuminator+unit/Maximum+Magnification+Module/pmc07041449-353-10-11
Average 95 stars, based on 1 article reviews
manual nikon tirf illumination module - by Bioz Stars, 2026-11
95/100 stars
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a Principles of STAR microscopy. The evanescent field decay is wavelength dependent, and STAR is based on the changing fluorescence ratio of two spectrally separated fluorophores. STAR imaging of CLCa dual-tagged with iRFP713 and EGFP can be used to study the curvature of clathrin-coated structures. b Current models describing formation of clathrin-coated vesicles (CCVs): i, Constant Curvature Model (CCM); ii, Flat-to-curved transition (FTC); iii, Flat clathrin lattice (FCL) accumulation and dispersion. c 5 μm silica microsphere dual-tagged with Alexa488 and Alexa647, imaged in epifluorescence (E) and TIRF (T), T647/T488 ratio, and theoretical (Theor—blue) and measured (Meas—red) z-position as a function of distance from the center of the bead, mean ± SEM from 31 beads. d Cos-7 cell expressing CLCa-iRFP713-EGFP (STAR) probe imaged simultaneously with 488 and 647 TIRF excitation, arrows indicate example clathrin accumulations, scale bar = 20 μm. Representative of 13 cells and five independent repeats. e Western-blot analysis of STAR probe overexpression in Cos-7 cells. Representative blot from two independent replicates. f Plot of transferrin internalization over time for WT (black) and STAR probe expressing (magenta) Cos-7 cells. Circles and triangles indicate means from three independent replicates at 0, 5, 10, 15, and 20 min for WT Cos-7 ( n = 93, 93, 93, 93, 93 cells) and CLCa-STAR OE ( n = 89, 91, 93, 86, 92 cells). Black and magenta lines represent means ± SD. There was no main effect of STAR probe expression on TF-568 uptake by two-way ANOVA [Probe, Stimulation]—[ F (1, 4) = 0.7909, p = 0.42; F (2.132, 8.526) = 189.5, p < 0.0001]. Šídák’s multiple comparisons test, details in Supplementary Table . g Kymograph showing of clathrin accumulation (cyan) and curvature formation (fire), full arrow indicates vesicle formation, dashed arrow indicates flat clathrin lattice assembly. The curvature channel is translated 3 pixels down for better visualization, scale bar = 5 μm. h Quantitative GFP (cyan) and iRFP713 (magenta) intensity and ∆z (orange) traces for representative CCV and FCL, darker shaded band—SD of detected signal. Lighter cyan, magenta, and orange indicate background mean and 2*SD above background, red dashed boxes indicate when GFP signal is significantly higher than threshold. a , b were created with BioRender.com.

Journal: Nature Communications

Article Title: Imaging vesicle formation dynamics supports the flexible model of clathrin-mediated endocytosis

doi: 10.1038/s41467-022-29317-1

Figure Lengend Snippet: a Principles of STAR microscopy. The evanescent field decay is wavelength dependent, and STAR is based on the changing fluorescence ratio of two spectrally separated fluorophores. STAR imaging of CLCa dual-tagged with iRFP713 and EGFP can be used to study the curvature of clathrin-coated structures. b Current models describing formation of clathrin-coated vesicles (CCVs): i, Constant Curvature Model (CCM); ii, Flat-to-curved transition (FTC); iii, Flat clathrin lattice (FCL) accumulation and dispersion. c 5 μm silica microsphere dual-tagged with Alexa488 and Alexa647, imaged in epifluorescence (E) and TIRF (T), T647/T488 ratio, and theoretical (Theor—blue) and measured (Meas—red) z-position as a function of distance from the center of the bead, mean ± SEM from 31 beads. d Cos-7 cell expressing CLCa-iRFP713-EGFP (STAR) probe imaged simultaneously with 488 and 647 TIRF excitation, arrows indicate example clathrin accumulations, scale bar = 20 μm. Representative of 13 cells and five independent repeats. e Western-blot analysis of STAR probe overexpression in Cos-7 cells. Representative blot from two independent replicates. f Plot of transferrin internalization over time for WT (black) and STAR probe expressing (magenta) Cos-7 cells. Circles and triangles indicate means from three independent replicates at 0, 5, 10, 15, and 20 min for WT Cos-7 ( n = 93, 93, 93, 93, 93 cells) and CLCa-STAR OE ( n = 89, 91, 93, 86, 92 cells). Black and magenta lines represent means ± SD. There was no main effect of STAR probe expression on TF-568 uptake by two-way ANOVA [Probe, Stimulation]—[ F (1, 4) = 0.7909, p = 0.42; F (2.132, 8.526) = 189.5, p < 0.0001]. Šídák’s multiple comparisons test, details in Supplementary Table . g Kymograph showing of clathrin accumulation (cyan) and curvature formation (fire), full arrow indicates vesicle formation, dashed arrow indicates flat clathrin lattice assembly. The curvature channel is translated 3 pixels down for better visualization, scale bar = 5 μm. h Quantitative GFP (cyan) and iRFP713 (magenta) intensity and ∆z (orange) traces for representative CCV and FCL, darker shaded band—SD of detected signal. Lighter cyan, magenta, and orange indicate background mean and 2*SD above background, red dashed boxes indicate when GFP signal is significantly higher than threshold. a , b were created with BioRender.com.

Article Snippet: Live-cell STAR image acquisition was performed with a Nikon Ti-2 microscope equipped with a motorized stage, stage-top incubator to maintain 37 °C and 5% CO 2 (Tokai Hit, INUBG2SF-TIZB), ×60 1.49-NA objective, manual TIRF illuminator (Nikon, TI-LA-TIRF), 488 nm (Obis, 488-150 LS), and 647 nm (Obis, 1196627) excitation lasers, fiber coupling optics: fiber mount (Thorlabs, MBT621D), converging and directing the laser objective (Olympus, RMS10X), optical fiber (Thorlabs, P3-405BPM-FC-2), C-NSTORM QUAD 405/488/561/638 nm TIRF dichroic.

Techniques: Microscopy, Fluorescence, Imaging, Dispersion, Expressing, Western Blot, Over Expression