[Histonet] MacNeal's tetrachrome

Gayle Callis gayle.callis <@t> bresnan.net
Wed Aug 13 11:32:29 CDT 2008

I will forward the method to you.  Do NOT purchase this stain as a 
commercial preparation.  It works best at an inhouse preparation. You will 
need to try the stain on the bone, as timing can vary with age of patient be 
it human or animal, also species, and bone type.  HOever if you are doing LR 
white, just how thick are the sections?   We did this stain on thick, slab 
sections as a surface stain.  You may not need to etch the bone for a really 
thin section (?)

Etching is done by immersing the bone into 1% formic acid (made from from 
stock 88%), for 30 sec to 1 min.  Using a sonicator inproves etching, but 
the section must be rinsed well after etching with running tap water, then 
dried followed by staining.  If the section is super thin, then timing can 
be reduced plus concentration of the formic acid to 0.5%.

I will forward to you after this message.

Gayle M. Callis
Bozeman MT

----- Original Message ----- 
From: "Orla Gallagher" <o.m.gallagher <@t> sheffield.ac.uk>
To: <histonet <@t> lists.utsouthwestern.edu>
Sent: Wednesday, August 13, 2008 10:01 AM
Subject: [Histonet] MacNeal's tetrachrome

> Dear Histonetters,
> Our lab would like to try staining some undecalcified LR White-
> embedded bone sections with MacNeal's tetrachrome stain. I've
> found a recipe on the Histonet archive which mentions making up
> the stain from scratch but doesn't say how long to stain for or how
> to treat the sections after staining. The recipe also mentions
> etching in formic acid before staining. Has anyone got some advice
> as I haven't tried any etching techniques before?
> Many thanks,
> Orla
> ******************************
> Ms. Orla Gallagher
> Academic Unit of Bone Biology
> D Floor (DU20) Medical School
> Henry Wellcome Laboratories for Medical Research
> University of Sheffield
> Beech Hill Road
> Sheffield S10 2RX
> www.sheffield.ac.uk
> E-mail:o.m.gallagher <@t> sheffield.ac.uk
> Tel:   0114 271 3783(lab)
>       0114 271 3337(office)
> Fax:   0114 271 1711
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