[Histonet] Muscle striations on paraffin - Sarcomeres

From:"Carlos Augusto"

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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