Nissl's Methylene Blue
for Nissl bodies
Materials
- Cajeput oil
- Staining solution
Material Amount 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.
- Differentiator
Material Amount 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.
Protocol
- Place sections in the staining solution in a watch glass.
- Heat gently until bubbles appear.
- Transfer to the differentiating fluid in another watch glass.
- Differentiate until color ceases to be extracted.
- Transfer the section to a slide and gently blot dry.
- Clear with cajeput oil.
- Mount with a resinous medium.
Expected Results
- Nissl – blue
- Nuclei – blue
Notes
- Nissl specified “Venetian soap”, which is the same as “Castile soap”. It refers to solid, bar soap made from olive oil and sodium hydroxide.
Safety Note
Prior to handling any chemical, consult the Safety Data Sheet (SDS) for proper handling and safety precautions.
References
- 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.
