Lectin-biotin assay for slime present in in situ biofilm produced by Staphylococcus epidermidis using transmission electron microscopy (TEM)

B. A. Sanford, V. L. Thomas, S. J. Mattingly, M. A. Ramsay, M. M. Miller

Research output: Contribution to journalArticlepeer-review

23 Scopus citations

Abstract

A lectin-biotin assay was developed for use in the specific detection of slime produced by Staphylococcus epidermidis RP62A and M187sp11 grown in a chemically defined medium. Mature biofilm was formed on polyvinylchloride (PVC) disks using a combined chemostat-modified Robbins device (MRD) model system. Specimens fixed in situ were: 1) stained with ruthenium red; 2) reacted overnight with biotin-labeled lectins (WGA, succinyl-WGA, Con A, or APA) followed by treatment with gold-labeled extravidin; or 3) reacted with antibodies against S. epidermidis RP62A capsular polysaccharide/adhesin (PS/A) using an immunogold procedure. WGA and succinyl-WGA (S-WGA), which specifically bind N-acetylglucosamine, were shown by TEM to react only with slime, both cell-associated and exocellular. In contrast, Con A, APA and anti-PS/A reacted with the bacterial cell surface but did not react with slime. These results indicate the usefulness of WGA lectin as a specific marker for detection of the presence and distribution of slime matrix material in S. epidermidis biofilm.

Original languageEnglish (US)
Pages (from-to)156-161
Number of pages6
JournalJournal of Industrial Microbiology
Volume15
Issue number3
DOIs
StatePublished - Sep 1995

Keywords

  • biofilm
  • lectin marker
  • S. epidermidis
  • slime

ASJC Scopus subject areas

  • Applied Microbiology and Biotechnology

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