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FP5

POLLINATOR PARASITES Informe resumido

Project ID: QLK5-CT-2002-00741
Financiado con arreglo a: FP5-LIFE QUALITY
País: Sweden

Improved protocols for TEM analysis of terrestrial Microsporidia in insects

Ultrastructural morphology of terrestrial Microsporidia is often difficult to describe because fixations and sectioning for transmission electron microscopy does not always yield optimal results. To improve our descriptions of pathology, gross morphology and ultrastructure of Microsporidia, needed species descriptions in this group efforts will be made to improve the standard protocol used in the project:

For transmission electron microscopy, infected tissue to be embedded for TEM will be fixed using 2.5 % glutaraldehyde (v/v) in 0.2M sodium cacodylate buffer, pH 7.4, for 12 - 48 h and kept refrigerated (+7 to 8oC) during prefixation. The specimens will be post-fixed for 2h in 2 % OsO4 (w/v) in 0.1 M S-colloidine buffer after washing in Na-cacodylate buffer.

The tissue pieces are then dehydrated in a series of ethanol solutions of ascending concentration, followed by fixation/washing/soaking in a propylene oxide solution. After dehydration, the tissue pieces are then embedded in Epoxy resin (Agar 100) by routine procedures for TEM.

Reported by

Department of Cell and Organism Biology, University of Lund
Helgonavaegen 3
223 62 Lund
Sweden
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