Nissl's Methylene Blue
for Nissl bodies


Cajeput oil
Staining solution
Castile soap 1.75 g
Methylene blue 3.75 g
Distilled water 1 L
Dissolve the soap in the water.
Add and dissolve the dye.
Allow to ripen at least three months
Aniline 10 mL
Ethanol, 95% 90 mL

Tissue sample
Ethanol fixation is preferred. Formalin fixed tissue may be suitable. Sections should be thicker than usual, and were originally to be free floating sections, likely free hand sections prepared without freezing.


  1. Place sections in the staining solution in a watch glass.
  2. Heat gently until bubbles appear.
  3. Transfer to the differentiating fluid in another watch glass.
  4. Differentiate until colour ceases to be extracted.
  5. Transfer the section to a slide and gently blot dry.
  6. Clear with cajeput oil.
  7. Mount with a resinous medium.

Expected results


  1. Nissl specified "Venetian soap", which is the same as "Castile soap". It refers to solid, bar soap made from olive oil and sodium hydroxide.


Wikipedia articles   Castile soap

Gray, Peter. (1954)
The Microtomist's Formulary and Guide. pp. 446.
Originally published by:– The Blakiston Co.
Republished by:– Robert E. Krieger Publishing Co.

Gatenby, J.B. and Beams, H.W., (1950)
The Microtomist's Vade-Mecum. 11 ed., pp. 508, para. 1099
Churchill, London, UK.




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