Confocal Laser Scanning Microscopy Is Appropriate to Detect Viability of Enterococcus faecalis in Infected Dentin

Ronald Ordinola Zapata, Clovis M. Bramante, Ivaldo Gomes de Moraes, Norberti Bernardineli, Thais Helena Gasparoto, Marcia S.Z. Graeff, Ana Paula Campanelli, Roberto Brandão Garcia

Research output: Contribution to journalArticlepeer-review

88 Scopus citations

Abstract

The purpose of this study was to explore the potential of confocal laser scanning microscopy (CLSM) for in situ identification of live and dead Enterococcus faecalis in infected dentin. Eight cylindrical dentin specimens were infected with Enterococcus faecalis in BHI for 21 days. After the experimental period, the specimens were stained with fluorescein diacetate (FDA) and propidium iodide (PI) or acridine orange (0.01%) and analyzed by CLSM. Two noninfected dentin specimens were used as negative controls. CLSM analysis shows that the discrimination between viable (green) and dead (red) bacteria in infected dentinal tubules could be observed after staining with FDA/PI. Acridine orange was able to show metabolic activity of the E. faecalis cells inside the dentinal tubules showed by its red fluorescence. The viability of bacteria in infected dentin can be determined in situ by CLSM. FDA/PI and acridine orange are useful for this technique.

Original languageEnglish (US)
Pages (from-to)1198-1201
Number of pages4
JournalJournal of Endodontics
Volume34
Issue number10
DOIs
StatePublished - Oct 2008
Externally publishedYes

Bibliographical note

Funding Information:
Supported by FAPESP (2007/01838-7).

Copyright:
Copyright 2009 Elsevier B.V., All rights reserved.

Keywords

  • Confocal laser scanning microscopy
  • Enterococcus faecalis
  • dentin
  • viability

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