This is a general protocol adapted from a protocol on ihcworld.com.
Purpose:Nisslstaininguses a cresyl violet solution to stainRNA, which is most abundant in the rough endoplasmic reticulum of nuclei. This method is used to detect the nuclei of neurons in fixed, embedded, and frozen tissue.
30-50 μm fixed frozen/vibratome sections, mounted on slides
Beakers to mix and hold solutions
Cresyl fast violet
Glacial acetic acid
100% ethyl alcohol
95% ethyl alcohol / 5% deionized H2O mixture
1)Prepare 0.1% cresyl violet solution by mixing 0.1 g cresyl fast violet mixed in 100 mL deionized H2O.Add 10 drops of glacial acetic acid just before use and filter.
2)De-fat the tissues by soaking in a 1:1 alcohol/chloroform mixture overnight.
3)Rehydrate the slices in 100% alcohol followed by a 95% alcohol / 5% deionized H2O mixture.
Note:Putting the slides in pure water would cause the frozen tissue to come off the slides.
4)Stain in cressyl violet for 3-5 minutes.
5)Rinse in distilled water.
6)Soak in 95% ethyl alcohol for 5-30 minutes.Check microscopically forstaining.
7)Dehydrate in 100% alcohol for five minutes.Replace alcohol and repeat.
8)Clear in xylene for five minutes.Replace xylene and repeat.
9)Mount with resin medium.
Check forstaining.Nissl bodies (neurons) will be stained red-violet.