RE: Thionin stain
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)
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.
-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
1124 Columbia St. Suite 650
Seattle, WA 98104
Direct: (206) 621-5120
Main: (206) 839-0300
Fax: (206) 839-0303
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