Abstract
The gram-positive coagulase-negative staphylococci are opportunistic organisms important in latent infection associated with prosthetic biomaterials. Their highly charged polysaccharide glycocalyx, or slime, aids in proliferation of bacteria on surfaces after an initial adhesion event, possibly at the time of surgical insertion. The cationic reagents, ruthenium red (RR) and alcian blue (AB), in en bloc procedures have aided visualization of the polysaccharide glycocalyx material in TEM or SEM applications. Further, enhancement of RR preservation or staining of slime by inclusion of lysine in a prefixation stage has been observed. The effect of lysine with and without RR and AB on preservation/staining of the staphylococci glycocalyx is investigated further in this study.Staphylococcus aureus ATCC 25923, Staphylococcus epidermidis RP62 (RP62) and Staphylococcus hominis SP2 (SP2) were cultured in trypticase soy broth for 18 hrs at 35°C, following recovery from frozen storage and plating on blood agar plates for 24 hrs. Cells with 75mM lysine in the prefixation were incubated for 20 minutes with 2.5% glutaraldehyde (GA), 2.5% GA and 0.075% RR, or 2.5% GA and 0.075% AB, respectively.
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Schedule a callMore From: Proceedings, annual meeting, Electron Microscopy Society of America
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