RE: Thionin stain

From:Stephanie Rodriguez

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 


NOTICE:  This communication may contain privileged or other confidential information intended for the addressee only. If you have received it in error, please advise the sender by reply email and immediately delete the message and any attachments without copying or disclosing the contents. Thank you.

<< Previous Message | Next Message >>