Immunohistochemistry of Paraformaldehyde-Fixed Tissue

From paraformaldehyde-fixed tissue, 7 μm sections were prepared, as previously described (Gray et al., 2012 (link)). In some cases, sections were fixed by acetone (Gray et al., 2012 (link)). Sections were stained with the flowing antibodies: anti-Lyve1-A647, anti-CD3ε-bio (clone 145-2C11), Goat anti-CCL20 (AF760), Polyclonal Rabbit anti-GFP (Thermo Fisher Scientific), anti-B220-A647 (RA3-6B2), anti-ICAM1-bio (3E2), anti-IL7Rα-A647 (A7R34), rat anti-scart2, Hamster anti-TCRγδ (GL3), Donkey anti-Goat-biotin Jackson immunoresearch), Goat anti-Armenian Hamster-bio (Jackson immunoresearch), Donkey anti-Rat-bio (Jackson immunoresearch), Anti-biotin-A488 (Jackson immunoresearch), Anti-biotin-Cy3 (Jackson immunoresearch). Images were captured with a Zeiss AxioOberver Z1 inverted microscope.

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Zhang Y., Roth T.L., Gray E.E., Chen H., Rodda L.B., Liang Y., Ventura P., Villeda S., Crocker P.R, & Cyster J.G. (2016). Migratory and adhesive cues controlling innate-like lymphocyte surveillance of the pathogen-exposed surface of the lymph node. eLife, 5, e18156.

Publication 2016

Polyclonal Rabbit anti-GFP Donkey anti-Goat-biotin AxioOberver Z1 inverted microscope

Acetone Antibodies Armenian hamster Biotin Ccl20 Cd3ε Clone Donkey Goat Hamster Icam1 Il7rα Microscope Paraformaldehyde Rabbit Tissue

Corresponding Organization : University of California, San Francisco

Other organizations : University of Washington, Seattle University, Broad Center, University of Dundee

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Variable analysis

independent variables

Tissue fixation method (paraformaldehyde or acetone)

dependent variables

Immunohistochemical staining of tissue sections with various antibodies (anti-Lyve1-A647, anti-CD3ε-bio, anti-CCL20, anti-GFP, anti-B220-A647, anti-ICAM1-bio, anti-IL7Rα-A647, anti-scart2, anti-TCRγδ)

control variables

Tissue section thickness (7 μm)
Microscope used for imaging (Zeiss AxioOberver Z1 inverted microscope)

controls

No positive or negative controls were explicitly mentioned in the provided information.

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