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. 2004;39(3):301-5.
doi: 10.1111/j.1472-765X.2004.01589.x.

A fluorescence-based method for the detection of adhesive properties of lactic acid bacteria to Caco-2 cells

M A Bianchi  1 , D Del Rio, N Pellegrini, G Sansebastiano, E Neviani, F Brighenti
Affiliations

A fluorescence-based method for the detection of adhesive properties of lactic acid bacteria to Caco-2 cells

M A Bianchi et al. Lett Appl Microbiol. 2004.

Abstract

Aims: The ability of probiotic micro-organisms to adhere to the intestinal surface is regarded as a substantial advantage in terms of bacteria persistence in the gastrointestinal tract. The aim of the present study was the development of a method based on fluorescent staining of bacteria and subsequent spectrofluorimetric detection to quantify the adhesion of several strains of Lactobacillus and Bifidobacterium to Caco-2 cells.

Methods and results: Lactic acid bacteria strains were subjected to fluorescent staining using the viable probe carboxyfluorescein diacetate and subsequently incubated on Caco-2 monolayers. The adhesion of the micro-organisms was determined by spectrofluorimetry following the lysis of the attached bacterial cells and expressed as adhesion percentage. The values obtained for the micro-organisms tested ranged from 4% for Bifidobacterium infantis Bi1 to 10% for a Bifidobacterium mixture containing three different strains.

Conclusions: In the present study we successfully applied fluorescent labelling and fluorimetric detection to investigate the adhesive properties of some Lactobacillus and Bifidobacterium strains and a Bifidobacterium mixture to Caco-2 cells.

Significance and impact of the study: The results proved that fluorescent labelling is suitable for adhesion studies and provides a reliable and safer alternative to radioactive labelling.

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