[Histonet] Weigert myelin stain procedure

Kemlo Rogerson Kemlo.Rogerson <@t> waht.swest.nhs.uk
Fri Sep 14 02:29:33 CDT 2007

Add 1% calcium chloride to 10% formalin and neutralise with suspended
calcium carbonate. Embed tissue in 25% gelatine then cool in a fridge
and then harden 1 day in 1% calcium chloride, 1% cadmium chloride, 10%
formalin solution- cut sections 15u, attach to slides (by floating onto
previously coated slides with 2.5% gelatine), drain then dry. Expose to
concentrated formalin to harden gelatine, and put back into calcium,
cadmium, formalin solution until needed.

Wash slides 3 min in water. Place in cold 5% potassium dichromate in a
Coplin jar then place in a 60 degree oven for 48 hours; allow jar to
cool. Wash slides in water(dist) then stain in modified Kultschitzky's
haematoxylin (haematoxylin 1g, dist water 98 ml, sodium iodate 0.2 g,
glacial acetic acid 2ml) at 37 degrees for 5 hours. The haematoxylin
forms a resistant black lake with the chromium held by the lipids.
Differentiate for 15 hours in Weigert's 1% borax, 2.5% potassium
ferrocyanide; only 8 hours for Golgi substance. Wash 5 min in water
mount in glycerol gelatine.

Cephalin and sphingomyelin are stained black, whilst cerebrosides are
grey. If memory serves sections fall off and you eventually lose the
will to live but if it works then it's rather pretty.

Kemlo Rogerson
Pathology Manager
DD   01934 647057 or extension 3311
Mob 07749 754194; Pager 07659 597107;

The world breaks everyone, and afterward, some are strong at the broken
places. --Ernest Hemingway 

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