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Applied and Environmental Microbiology, August 1998, p. 3059-3062, Vol. 64, No. 8
0099-2240/98/$00.00+0

Estimation of the State of the Bacterial Cell Wall by Fluorescent In Situ Hybridization

Elena Bidnenko,1 Carine Mercier,2 Josselyne Tremblay,2 Patrick Tailliez,2 and Saulius Kulakauskas2,*

Génétique Microbienne1 and Unité de Recherches Laitières et de Génétique Appliquée,2 Institut National de la Recherche Agronomique, Domaine de Vilvert, 78352 Jouy-en-Josas, France

Received 20 January 1998/Accepted 8 June 1998

Fluorescent in situ hybridization (FISH) is now a widely used method for identification of bacteria at the single-cell level. With gram-positive bacteria, the thick peptidoglycan layer of a cell wall presents a barrier for entry of horseradish peroxidase (HRP)-labeled probes. Therefore, such probes do not give any signal in FISH unless cells are first treated with enzymes which hydrolyze the peptidoglycan. We explored this feature of FISH to detect cells which have undergone permeabilization due to expression of autolytic enzymes. Our results indicate that FISH performed with HRP-labeled probes provides a sensitive method to estimate the states of cell walls of individual gram-positive bacteria.

* Corresponding author. Mailing address: Unité de Recherches Laitières et de Génétique Appliquée, Institut National de la Recherche Agronomique, Domain de Vilvert, 78352 Jouy-en-Josas, France. Phone: (33) 01 34 65 20 73. Fax: (33) 01 34 65 20 65. E-mail: kulakaus{at}biotec.jouy.inra.fr.

Applied and Environmental Microbiology, August 1998, p. 3059-3062, Vol. 64, No. 8
0099-2240/98/$00.00+0


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