Buffered Thionin Stain
for Nissl Bodies

Solutions

Staining solution pH 3.7 pH 4.5
Acetic acid, 0.6% (0.1M) 90 mL 60 mL
Sodium acetate, 0.8% (0.1M) 10 mL 40 mL
Thionin, 1% aqueous 2.5 mL 2.5 mL

Tissue sample
10µ paraffin sections fixed in 10% formalin variants or Carnoy's chloroform-ethanol-acetic mixture are suitable. Other fixatives may be satisfactory. x

Method

  1. Bring sections to water via xylene and ethanol.
  2. Place into one of the staining solutions for 20-60 minutes.
  3. Dehydrate with ascending concentrations of ethanol.
  4. Clear with xylene and mount with a resinous medium.

Expected results

Structure  –  pH 3.7 pH 4.5
Nissl bodies  –  blue dark blue
Nuclei  –  blue dark blue
Background  –  pale or unstained pale blue

 

Notes

  1. Alternatively:
    1. Dilute the thionin with distilled water instead of acetate buffer.
    2. Bring sections to water via xylene and ethanols.
    3. Stain in aqueous thionin for 20-60 minutes.
    4. Rinse with ethanol, 50%.
    5. Differentiate with 0.25% acetic acid in 95% ethanol, controlling microscopically.
    6. Rinse well with 95% ethanol.
    7. Complete dehydration with absolute ethanol.
    8. Clear with xylene and coverslip using a resinous medium.

 

Reference
Davenport, H.A.. (1960).
Histological and Histochemical Technics,
W. B. Saunders, Philadelphia, USA.
Citing:
 Windle, W. F., Rhines, R. and Rankin, J., , (1943),
 A Nissl method using buffered solutions of thionin.
 Stain Technology, v 8, pp. 77-86.
and:
 Conn, H. J. and Darrow, M. A.,, (1946),
 Staining procedures.
 Biotech Publications, Geneva, New York.

 


 

 

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