E3 Journal of Biotechnology and Pharmaceutical Research

E3 Journal of Biotechnology and Pharmaceutical Research Vol. 4 (4) pp. 61-67, April 2013; © E3 Journals; ISSN 2141-7474

Fluorescence investigation of discriminatory phagocytosis of live Escherichia coli and Lactobacillus rhamnosus by J774 Murine macrophage

Abstract

To investigate the phagocytic behavior of J774 murine macrophages in the presence of Lactobacillus rhamnosus and Escherichia coli, the bacteria were cultured and labeled with Fluorescein Isothiocyanate (FITC) and Rhodamine6G fluorescent dyes respectively and incubated with the macrophages. On ingestion, fluorescence micrographs of the culture were captured using fluorescein and Rhodamine filters respectively and processed using the Openlab software. The resultant images were synchronized with Imagehopper2 and analyzed. The fluorescence micrographs revealed that Murine macrophages selectively internalized E.coli in the presence of the probiotic, Lactobacillus rhamnosus. The results suggest a molecular probiotic recognition mechanism in the mammalian immune system with potentially therapeutic applications and emphasize the increasing importance of fluorescence labeling methods in live imaging of in-vitro studies

Keywords: Discriminatory phagocytosis, phagocytosis assay, fluorescence microscopy