AW: [Histonet] decalcifiers and Safranin O staining

Gudrun Lang gu.lang <@t>
Wed Dec 19 13:44:26 CST 2007

HCl has a acid-constant of -6 (strong acid) and formic acid has 3,75 (medium
strong acid).
Strong acids saponificate amidgroups resulting in an acid-group,
deamination. Aminogroups are the reactionpartners of the collagen-stains.
Therefore they can't bind to a deaminated collagen. Another function of a
strong acid is to break down the peptidlinkage and generally denaturate a
protein. The three-dimensional configuration of collagen could also
influence the staining affinity.

Doesn't Immunocal consist of formic acid and formaldehyd? I consider, that
the deamination-effect and the break down appear more on underfixed
specimen; and that formic acid isn't strong enough to disturb the
collagen-configuration in the same manner.
My thoughts about it, but no academic evidence.

Gudrun Lang
Biomed. Analytikerin
Akh Linz
Krankenhausstr. 9
4020 Linz
-----Ursprüngliche Nachricht-----
Von: histonet-bounces <@t>
[mailto:histonet-bounces <@t>] Im Auftrag von Michele
Gesendet: Mittwoch, 19. Dezember 2007 19:24
An: histonet <@t>
Betreff: [Histonet] decalcifiers and Safranin O staining

Can anyone explain the chemistry behind decalcifying agents and the
Safranin O procedure?...Specifically, why does using Immunocal (formic
acid) as a decalcifier yield beautiful Safranin O staining while RDO
(HCL) almost totally eliminates staining? Does HCL do something to the
proteoglycans that formic acid does not?




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