[Histonet] Losing sections.

[Alicia Sedo]

Hello Histonet,

I've been trying a Van Gieson stain on cryosections of mouse cardiac tissues,
that have previously been stained for lacZ expression and are sitting in 4% PBS
Formaldehyde.

I have been losing the sections from the slides at the Van Gieson solution step.
I am omitting the nuclear stain, instead going from washing in tap water to
rinsing in distilled, then into the Van Gieson.
I got the protocol (Staining of Collagen- on Frozen sections) from Histonet.
The stain is 0.5% Aqueous Acid Fuchsin-15ml, Saturated Picric Acid-75ml,
Distilled water-50ml...with 250microlitres of concentrated Nitric acid, which
was recommended to sharpen the differentiation of the colours.

The sections lift off and curl up after a few seconds, the slides are homemade
silane coated and have withstood all other procedures so far. I have tried the
stain without the Nitric acid but it still happens. I have previously tried
this Van Gieson on FFPE cardiac tissue on the same slides, with good results.

Am I missing something obvious that I don't know about? I am not a Histologist
only a researcher in an University lab. I hope someone out there can point out
what I am doing wrong!

Alicia Sedo.

Cardiovascular Research
Level 11 Worsley Building,
Clarendon Way
University of Leeds.
LS2-9JT.