Ovarian Tissue Fixation and X-gal Staining

As described [25 (link)], ovarian samples were first fixed in 2% paraformaldehyde-0.2% glutaraldehyde in a 0.1 M phosphate buffer solution (pH 7.4), and then stained with X-gal staining buffer containing 1 mg/ml X-gal, 5 mM potassium ferricyanide, and 5 mM potassium ferrocyanide. After the staining procedure, samples were post-fixed with 10% neutral buffered formalin for histological analysis. Fast red (Vector Laboratories) was used to counterstain the nucleus.

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Gao Y., Fang X., Vincent D.F., Threadgill D.W., Bartholin L, & Li Q. (2017). Disruption of postnatal folliculogenesis and development of ovarian tumor in a mouse model with aberrant transforming growth factor beta signaling. Reproductive Biology and Endocrinology : RB&E, 15, 94.

Publication 2017

Fast red

Formalin Glutaraldehyde Nucleus Ovarian Paraformaldehyde Phosphate Potassium ferricyanide Potassium ferrocyanide Vector X gal

Corresponding Organization :

Other organizations : Texas A&M University, Cancer Research UK Scotland Institute, Cancer Research UK, Centre de Recherche en Cancérologie de Lyon, Université Claude Bernard Lyon 1, Centre National de la Recherche Scientifique, Centre Léon Bérard, Inserm

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Variable analysis

independent variables

Fixation of ovarian samples in 2% paraformaldehyde-0.2% glutaraldehyde in a 0.1 M phosphate buffer solution (pH 7.4)
Staining of samples with X-gal staining buffer containing 1 mg/ml X-gal, 5 mM potassium ferricyanide, and 5 mM potassium ferrocyanide
Post-fixation of samples with 10% neutral buffered formalin for histological analysis
Counterstaining of the nucleus with Fast red (Vector Laboratories)

dependent variables

Staining/color intensity of the samples

control variables

PH of the phosphate buffer solution (7.4)
Concentrations of fixatives (2% paraformaldehyde, 0.2% glutaraldehyde)
Concentrations of X-gal, potassium ferricyanide, and potassium ferrocyanide in the staining buffer
Concentration of neutral buffered formalin for post-fixation

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

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