RE: [Histonet] Muscle striations on paraffin - Sarcomeres

From:"Tony Henwood"



Have you tried a PTAH stain?

Regards

Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC)
Laboratory Manager & Senior Scientist
Tel: 612 9845 3306
Fax: 612 9845 3318
the children's hospital at westmead 
Cnr Hawkesbury Road and Hainsworth Street, Westmead 
Locked Bag 4001, Westmead NSW 2145 




-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Carlos Augusto
Sent: Wednesday, 13 August 2008 6:21 AM
To: histonet@lists.utsouthwestern.edu
Subject: [Histonet] Muscle striations on paraffin - Sarcomeres


Dear histonetters

Here at the lab a university doctor have asked to do some muscle histology of scallop and oyster species.

She asked if the striations could be looked in a Masson´s Trichrome stain. She intends to measure the sarcomeres (the spaces between two z-lines) in the mollusc´s muscles, but I have had a lot of problems when the sections are stained.

1- In other species the striations are clearly visible (rabit, fish and chicken slides from the student´s histology couse were stained at the same time), but not in the molluscs.
2- Cells (muscle fibers) are very thin and even in 4 micrometer sections the fibers one over the others. So the microscopy is very "disrty" to look at.
3- We changed the stain for the Millingan´s Trichrome but only in one or two slides could be seen some signs of striation.


The tissues are fixed in 10% Neutral Buffered Formalin, washed, dehydrated in ethylic alcohols, cleared on hemo-de (xylol substitute-limonenes), infiltrated and embbeded with paraplast. Sections were done on a rotary microtome with disposable blades.


Any other way to look at the sarcomeres on paraffin processed tissues are very VERY appreciated.


Is there one way to know if there is a stain to differentiate sriated from smooth muscle cells?



Thanks in advance.
Carlos Aguilar Cruz
Laboratorio de Histología - Unidad Pichilingue
Universidad Autónoma de Baja California Sur.

Carretera al Sur KM 5.5 CP230080
La Paz, BCS, México

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