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Buffered Thionin for Nissl Bodies


Buffered Thionin

for Nissl Bodies



MaterialAmount for pH 3.7 SolutionAmount for pH 4.5 Solution
Acetic acid, 0.6% (0.1M)90mL60mL
Sodium acetate, 0.8% (0.1M)10mL40mL
Thionin, 1% aqueous2.5mL2.5mL

Tissue Sample

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


Standard 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.

Alternative Method

  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.

Expected Results

StructurepH 3.7 Staining SolutionpH 4.5 Staining Solution
Nissl bodiesbluedark blue
Nucleibluedark blue
Backgroundpale or unstainedpale blue

Safety Note

Prior to handling any chemical, consult the Safety Data Sheet (SDS) for proper handling and safety precautions.


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