首页 » 实验方法 » 神经科学技术 » 神经科学技术

Nissl Staining for Neurons

来源:生物谷  2009-3-11   访问量:8394
评论(0)
分享

Description

From  the lab of Lucien T. "Tres" Thompson, Ph.D. The University of Texas at Dallas

 

Nissl Staining for Neurons

 

This is a general protocol adapted from a protocol on ihcworld.com.

 

Purpose:  Nissl staining uses a cresyl violet solution to stain RNA, 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.

 

Materials

            30-50 μm fixed frozen/vibratome sections, mounted on slides

            Beakers to mix and hold solutions

            Incubation chamber

Solutions

            Cresyl fast violet

            Glacial acetic acid

            100% ethyl alcohol

            Chloroform

            95% ethyl alcohol / 5% deionized H2O mixture

            Xylene

            Resin medium

 

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 for staining.

 

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 for staining.  Nissl bodies (neurons) will be stained red-violet.

相关神经科学技术