I want to stain some of my mouse brain sections with Cresyl Violet.
The mouse brains (not perfused, no PFA/Formalin used) were fresh frozen in OCT and cut in the cryostat at 15μm thickness. The sections were collected on normal slides and stored at -20℃.
Could you please help me with the staining protocol?
Here is a staining protocol for Cresyl Violet staining of fresh frozen mouse brain sections:
Materials:
Procedure:
- Prepare the Cresyl Violet solution according to the manufacturer's instructions or a standard protocol. Typically, a 0.1-0.5% Cresyl Violet solution in distilled water is used.
- Prepare 70%, 95%, and 100% ethanol solutions.
Note: Since the brain sections were not fixed with formalin, the staining may have some variability compared to fixed tissues. It is important to optimize the staining conditions, including the concentration and staining time with Cresyl Violet, based on the specific needs of your experiment.
Hi Priyadarshini Dutta
I agree with the above protocol, However there are a few modifications in mine.
Cresyl Violet Acetate Solution Preparation
1. Solution A: 6mL-glacial acetic acid + 994 mL- H2O
2. Solution B: 13.6g Sodium acetate + 1000mL H2O
3. Combine A and B solution in 9:1 ratio and adjust pH to 3.7.
4. Prepare 1% cresyl violet in the above solution. Filter and use the same for staining.
* Further as PFA fixation was not done, I would suggest to do fixation using ice cold methanol- 10mins followed by PBS wash.
* In addition to the above protocol I also perform differentiation post staining with cresyl violet (2 drops GAA in 95% ethanol).
* I use PBS for washing during the staining.
Do let me know if there are any queries.
Thanks
Samir Ranjan Panda Saif Wahid - Thank you a ton, for all your help.
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