Cresyl Violet Staining
This staining protocol is used on lymphoid tissue sections in order to conserve mRNA prior to laser microdissection of specific areas that will then be subjected to RT-PCR to assess gene expression in situ.
1) Preparation of Cresyl Violet solution
Dissolve 0.1 g of Cresyl Violet acetate (sigma Ref: C1791) in 10 ml of RNase free absolute Ethanol (1% W/V).
Leave over a rotator OVN to dissolve completely, and filter.
2) Preparation and staining of the tissue sections
Cut and acetone fix the sections as usual. Only use clean knives and acetone solution.
Rehydrate in absolute Ethanol, then 70% Ethanol, and 50% Ethanol. All RNAse free solutions.
Stain for 3min with cresyl violet.
Quickly dehydrate in 50% Ethanol, 70% Ethanol, absolute Ethanol.
Dip two/three times in each solution, not longer to avoid washing out the dye.
Leave it dry. Ready for microdissection !
Cresyl Violet Staining —> PDF