ISSN: 2329-8901
Dairy Science Building, LSU AgCenter, Baton Rouge, LA 70803, USA
Research Article
Cell Wall Integrity and Protoplast Formation of the Probiotic Lactobacillus acidophilus through Fluorescent Staining and Fluorescence Microscopy
Author(s): Ryan Page, David Burk and Kayanush Aryana*
The identification of protoplast of bacterial cells has previously utilized phase contrast microscopy. This method
determines protoplast by their size and change in shape. A more verifiable method can be used utilizing fluorescent
stains that target the specific cellular components. The goal of this study was to utilize fluorescence microscopy
techniques to determine the presence or absence of bacterial cell walls in the probiotic Lactobacillus acidophilus, after
exposure to cell wall digestive enzymes. Bacterial cells were treated with different concentrations of lysozyme [0, 175,
250, 425 μg/ml] and were incubated at 37°C for ten minutes. Following lysozyme treatment cells were fluorescently
stained with different concentrations (1x, 2x, 10x, and 100x) of two fluorescent dyes, Wheat-germ agglutinin (WGA)
and Hoechst 33342. The WGA [CF®.. View More»