[Histonet] Hematoxylin Puris

[Barry R Rittman]

Chris
Heidenhains iron hematoxylin is not specific for Entamoeba, it can be
used to shown anything from nuclei to muscle striations. The structures
demonstrated depend on the degree of differentiation. The stain useful
because of the intensity of staining due to the iron-hematoxylin that is
formed.
Regaud's hematoxylin is a modified Heidenhains and usually, because of
glycerol in the mixture gives a little more uniform of a stain.
Both depend on mordanting in iron alum followed by hematoxylin and then
differentiating in iron alum.
Not sure if Regaud's is commercially available but it is not that
difficult to make.
5% aqueous iron alum as mordanting and differentiating soln. (I prefer
to use a weaker iron alum solution for differentiation for better
control. When making this solution make sure that the crystals you use
are lilac in color as the crystals tend to have a yellow powder if kept
too long. I prefer to use iron alum solutions that are fresh although
the solution will keep for several weeks.
Hx stain is 1 gm hematoxylin, 10 ml. 90% ethanol, 80 ml. Distilled
water, 10 ml. glycerin. This need sto be some weaks old or else ripened
before use.
>From distilled water, mordant in iron alum soln. For 30 mins plus.
Rinse in several changes distilled water.
Stain in Regaud's soln. for 30-60 mins.
Better staining if you carry out the mordanting and staining at 50-60C.
Rinse in distilled water
Differentiate in iron alum.
Wash well in tap water etc.
Barry



Hello Histonetters,

 Can anybody help me out with a Heidenhain's Iron Hematoxylin Stain?
It's a special stain that directly demonstrates the parasite Entamoeba
Histolytica.
Can anybody guide me through where to purchase Regaud's Iron
Hematoxylin? One of the key reagents is hematoxylin puris?
Any help is appreciated.

Chris