Chicken Embryo Urogenital Tissue Histology

Urogenital tissues were dissected from chicken embryos and briefly fixed for 15 min in 4 % paraformaldehyde/PBS at room temperature. Tissues were then cryoprotected by immersion in 30 % sucrose/PBS (overnight at 4 °C), infiltrated with OCT embedding compound. Ten micron frozen sections on slides were treated as previously described [63 (link)]. Secondary antibodies were AlexFluor 488 donkey anti-rabbit IgG, 1:1000, and AlexaFluor 594 donkey anti-mouse IgG, at 1:1500 from Invitrogen). Sections were mounted in Fluorosave (Calbiochem). Images were taken on a Zeiss Axiovision M1.

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Ayers K.L., Lambeth L.S., Davidson N.M., Sinclair A.H., Oshlack A, & Smith C.A. (2015). Identification of candidate gonadal sex differentiation genes in the chicken embryo using RNA-seq. BMC Genomics, 16(1), 704.

Publication 2015

AlexFluor 488 donkey anti-rabbit IgG AlexaFluor 594 donkey anti-mouse IgG Axiovision M1

Anti igg Antibodies Chicken Donkey Embryos Frozen sections Immersion Mouse Paraformaldehyde Rabbit Sucrose Tissues Urogenital

Corresponding Organization :

Other organizations : University of Melbourne, Royal Children's Hospital, Murdoch Children's Research Institute, Monash University

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Variable analysis

independent variables

Tissue dissection from chicken embryos
Tissue fixation duration (15 min)
Tissue fixation concentration (4% paraformaldehyde/PBS)
Tissue cryoprotection (30% sucrose/PBS, overnight at 4°C)
Tissue infiltration with OCT embedding compound

dependent variables

Tissue morphology and structure (observed through frozen sections)
Protein expression/localization (detected by immunofluorescence labeling)

control variables

Temperature (room temperature for fixation, 4°C for cryoprotection)
Tissue sectioning thickness (10 microns)
Imaging platform (Zeiss Axiovision M1)

controls

Positive control: Not mentioned
Negative control: Not mentioned

Annotations

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