Re: shrinkage
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From: | Neuropathology <Neuropath.Frenchay@dial.pipex.com> |
To: | Histonet <histonet@pathology.swmed.edu> |
Reply-To: | |
Content-Type: | text/plain; charset=iso-8859-1 |
Having, due to gross carelessness, dropped a muscle biopsy into formalin I
have to disagree. When I hauled it back out, after a few minutes, the
fibres had shrunk considerably.
Andy Shand
----- Original Message -----
From: J. A. Kiernan <jkiernan@julian.uwo.ca>
To: Jennings, Margaret A. <jennings@mayo.edu>
Cc: Histonet <histonet@pathology.swmed.edu>
Sent: Sunday, August 27, 2000 2:47 AM
Subject: Re: shrinkage
> On Fri, 25 Aug 2000, Jennings, Margaret A. wrote:
>
> > What is the formula for shrinkage when using 10% neutral buffered
formalin
> > to fix a tissue sample?
>
> Aqueous formaldehyde (buffered or otherwise, with or without salts
> to make it isotonic) does not change the volume of a specimen. For
> details and references, read J.R.Baker's Principles of Biological
> Microtechnique, a great classic in the field of fixation and staining.
>
> > If I start with a tissue a microns x b microns x c
> > microns what do I end up with? Is there a time factor for additional
shrink?
>
> The shrinkage of a formaldehyde-fixed block occurs during dehydration,
> clearing, paraffin embedding and cutting and mounting the sections.
> Linear dimensions in the plane of the section are typically 70 to 75%
> of the the "fresh" sizes, with much variation among different organs.
>
> If you cut cryostat sections and collect them onto slides or coverslips
> they should be approximately life-size in the plane of the slide. The
> vertical dimension shrinks with drying. Thick frozen sections, variously
> processed before mounting onto slides, may go through large and
> uncontrolable size changes. Flattening the section onto the slide with
> a brush, prior to air-drying, can cause conspicuous expansion of the
> area of a section.
>
> The only way to know the amount of shrinkage is to measure the fresh
> specimen and its equivalent dimensions in the stained and mounted
> sections.
>
> > Is there also a formula for rate of penetration? Formalin infiltrates y
> > microns per t? What about other fixatives any formulas available?
>
> The formula for depth penetrated is useful only for fixatives that
> instantly cause a visible change as they diffuse into the specimen.
> These are the coagulants, such as alcohol and mercuric chloride.
> Baker's book says a lot about this, and provides data not published
> elsewhere, as far as I know. Baker explained, with examples, that
> in the case of formaldehyde the diffusion formula could not be
> used, because the small molecules penetrated specimens rapidly but
> reacted quite slowly to produce fixation, and the fixation did
> not cause a visible change like that brought about by coagulant
> fixatives.
>
> Specimens should be fixed in formaldehyde (especially if neutral
> and buffered, which slows down the reactions with protein) for
> at least 24 hours to obtain reasonable fixation. For full
> formalin fixation you need at least a week. Specimens dehydrated
> after only a few hours in formaldehyde are fixed principally by
> alcohol. Frozen sections cut after incomplete formaldehyde fixation
> are just that: incompletely fixed. Often that's what's wanted,
> especially in research work with the CNS.
>
> John A. Kiernan,
> Department of Anatomy & Cell Biology,
> The University of Western Ontario,
> LONDON, Canada N6A 5C1
>
>
>
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