Hi Tom,

We routinely use thionin to stain nissl bodies; we have also used LFB and CV in the past, but our researchers, too, prefer thionin, and brought their own recipe with them.

It is a very simple stain to perform, once you've made it up; I stain 14um mouse brain sections from water for 5 minutes, then differentiate with 2 changes 70% ethanol (about 10-20 dips), then 2 changes 95%, and dehydrate and clear with 3 changes each of 100% and xylene, respectively, and mount with resinous medium.

Here's the stain recipe:  (Some of the chemistry seems counterintuitive to me, but I haven't taken the time to improve on it)

Stock Solutions:

1.  1M acetic acid:  dilute 60mL glacial acetic acid in 940mL of dH2O.
2.  1N sodium hydroxide:  add dH2O to 40g NaOH pellets to a final volume of 1L.

Staining solution:

	-Add 36mL of stock NaOH solution to 200mL of stock acetic acid solution.  qs to 1L
	-Heat solution to about 60C, then slowly add thionin with stirring.  We use a 0.25% solution, so we add 2.5g thionin.
		(Thionin powder from Sigma cat# T-3387)
	-Cover and boil for 45 minutes.
	-Filter over #2 Whatman filter paper into a brown bottle to protect from light.
	-Stain is stored at RT and is good for about 4 months. 

Tissue stains in a similar fashion to staining with cresyl violet, but is a bluish-violet color under both light and dark-field microscopy.

If you have any other questions, you may email me directly.


Stephanie Rodriguez, B.S., HTL(ASCP) 
Neuroanatomy Lab Manager 
Research Associate III/Histotechnologist 
Primal, Inc. 
1124 Columbia St.  Suite 650 
Seattle, WA  98104 
Direct:  (206) 621-5120 
Main:  (206) 839-0300 
Fax:  (206) 839-0303 

 

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