Nematology , 2004, Vol. 6(3), 319-327 High-pressure freezing and freeze substitution of gravid Caenorhabditis elegans (Nematoda: Rhabditida) for transmission electron microscopy Myriam C LAEYS 1 , Dimitri V ANHECKE 2 , Marjolein C OUVREUR 1 , Tom T YTGAT 1 , August C OOMANS 1 and Gaëtan B ORGONIE 1 , ∗ 1 Ghent University, Department of Biologie, Ledeganckstraat 35, 9000 Ghent, Belgium 2 University Bern, Institute of Anatomy, Buehlstrasse 26, CH-3000 Bern 9, Switzerland Received: 22 September 2003; revised: 9 March 2004 Accepted for publication: 11 March 2004 Summary – Because chemical fixatives have a profound negative influence on tissue morphology and antigenicity, alternative fixation methods must be applied for some purposes. In this work we used high-pressure freezing (HPF) followed by freeze substitution to maximally preserve antigenicity and morphology. We developed a pipette method for bringing living Caenorhabditis elegans nematodes into the HPF recipient. Using cellulose tubes, it is possible to select individual nematodes for fixation. We were able to HPF complete adults and preserve the morphology in an enhanced fashion compared to chemically fixed tissue. Cellular organelles, especially mitochondria, were much better preserved. Uterine embryos protected by the intact eggshell were excellently preserved without the
Nematology – Brill
Published: Jan 1, 2004
Keywords: CRYO-TEM; ANTIGENICITY; ANTISERA; ULTRASTRUCTURE; MORPHOLOGY
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