Need help with GMA sectioning

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From:"Jacqueline F. Webb" <>
Date:Wed, 4 Aug 1999 16:42:35 -0500
Content-Type:text/plain; charset="us-ascii"

Hope someone out there can help.

We have been sectioning fish heads (portions of large ones or whole small
ones) for a study of sensory systems and swim bladder specializations in
butterflyfish.  These fish have a very thick stratum compactum in their
dermis, which is composed of fibrous collagen.  Infiltration has been good
(over 2 days), translucent tissue is the result. We also use a bit of
vacuum to get rid of bubbles. Polymerization is carried out in the frig for
larger blocks because of rising temperatures.  Sections at 5 um are fine,
but when we float them out on water (room temp or very warm) the plastic
stretches, the internal soft tissues stretch, but the collagen in the
dermis doesnt stretch. The result is ruffles radiating out from the tissue
and folds in the internal tissues. Staining and subsequent drying does not
change this situation.  This prevents any decent publishable photos from
being taken at lower magnifications (which is essential).  The tissue (a
marine fish) was fixed in 10% formalin in Seawater or in phosphate buffer
and decalcified (with radiographic confirmation) by Cal-Ex (Fisher)
overnight with 3-4 hours running tap water rinse.


Jackie Webb
Asst. Prof. Biology
Villanova University

Jacqueline F. Webb
Department of Biology
Villanova University
Villanova, PA  19085
610-519-7086 (office/lab)
610-519-7863 (FAX)

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