Improved sectioning and ultrastructure of bacteria and animal cells embedded in Lowicryl

Journal of Electron Microscopy Technique
J C BenichouA Ryter

Abstract

Lowicryl K4M-embedded Gram-positive and Gram-negative bacteria have a tendency to separate between the cell surface and the resin. This often leads to distortion of bacteria and more especially of mycobacteria. We describe attempts made to overcome this technical problem. Different assays were made on Bacillus subtilis, Escherichia coli, and Mycobacterium avium: 1) Modification of the bacterial surface by coating of bacteria with proteinic compounds; 2) treatment of bacteria with metallic salts known to modify cell wall polysaccharides; and 3) comparison between Lowicryl K4M and HM20. Conditions have been found in which the separation of all bacterial species from the resin is abolished. The most important factor appeared to be the treatment of bacteria before dehydration, with 0.5% uranyl acetate for 30 min. The second most important factor, especially for M. avium and to a lower extent for Gram-negative bacteria, was the use of Lowicryl HM20. No differences were observed with Gram-positive bacteria between K4M and HM20. Pre-embedding in gelatin instead of agar improved sectioning of M. avium, but had no effects on the other bacterial species. These conditions applied to macrophages infected with Shigella dysenteriae or M. avi...Continue Reading

References

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Citations

Nov 11, 1999·FEMS Microbiology Letters·J P Müller, M Wagner
Sep 1, 1996·Parasitology·M K JonesD P McManus
Aug 1, 1991·Zentralblatt Für Bakteriologie : International Journal of Medical Microbiology·N RastogiH L David
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Jan 28, 1999·Journal of Bacteriology·S LandaisH Sakamoto
May 1, 1992·Infection and Immunity·V AnnaixG Tronchin

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