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        Nissl Staining for Neurons

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        4428

        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 H2 O mixture

        Xylene

        Resin medium

        1) Prepare 0.1% cresyl violet solution by mixing 0.1 g cresyl fast violet mixed in 100 mL deionized H2 O. 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 H2 O 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.

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