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      Nissl Counterstaining - Cresyl Violet

      Ariadna Laguna, Miquel Vila, Núria Peñuelas

      Published: 2024-06-17 DOI: 10.17504/protocols.io.3byl49wmrgo5/v1
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      Abstract

      Nissl Counterstaining with Cresyl Violet for histological sections

      Steps

      Delipidation Step

      1.

      Ethanol 95%, 5min

      2.

      Chloroform 100%, 10min (under the hood)

      3.

      Ethanol 95%, 15 sec (quick rinse)

      4.

      Ethanol 95%, 2min

      5.

      Ethanol 70%, 1min30sec

      6.

      dH2O, 2min

      Cresyl Violet staining

      7.

      Cresyl Violet solution, 1min

      8.

      dH2O, 15sec or less (quick rinse)

      9.

      dH2O, 2min

      Differentiation

      10.

      Ethanol 70%+Acid Acetic*, few seconds (control differentiation yourself). (*add 2-3 drops

      of acetic acid d 100% in 100ml ethanol 70%).

      Dehydratation

      11.

      Ethanol 95%, 5-10sec (quick rinse)

      12.

      Ethanol 100%, 1min

      13.

      Xylene, 1min or more ( under the hood ) until mounting

      14.

      Mount coverslip with DPX ( under the hood )

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