R. Doyle, I. Ofek's Adhesion of Microbial Pathogens [no cover, index] PDF

By R. Doyle, I. Ofek

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Krogfelt, L. K. Poulsen, and S. Molin, Infect. Immun. 61, 5029 (1993). P. Panigrahi, G. T. DeTolla, and J. G. , Infect. Immun. 60, 4938 (1992). 52 G E N E R A L METHODS FOR A D H E S I O N TO A N I M A L CELLS [5] Procedure Covalent Binding of D-Mannose to Sepharose Beads" 1. 5 g of epoxy-activated Sepharose 6B (Pharmacia, Uppsala, Sweden) in 100 ml of distilled water for 15 min. 2. Wash the gel material thoroughly on a sintered glass funnel (pore size G 3) with a total of 350 ml of distilled water.

I. Gallin, W. M. Nauseet, and R. K. ), "LaboratoryManual of Neutrophil Function," p. 103. Raven, New York, 1986. 0 × 109 bacteria/ml Superoxide dismutase (Sigma) Catalase (Sigma) Procedure 1. One milliliter of the mast cell suspension is sedimented by centrifugation (600 g for 10 min), and the pellet is suspended in 900/~1 of luminol (final concentration, 10 ~M). 2. The mixture is transferred to a scintillation vial to which 100/A of the bacterial suspension is added. 3. The vial is swirled for 30 sec, placed into the/3-liquid scintillation counter, and counted for 1-min periods continuously for 30 min.

Although several techniques could be employed to study the adhesion of bacteria to mast cells, the method employing microscopy is particularly useful, because it reveals the characteristic patterns of bacterial adhesion on the mast cell surface as well as the heterogeneity in receptivity among mast cells. 2%, w/v) HBSS Procedure 1. One hundred microliters of the mast cell suspension is mixed with an equal volume of the bacterial suspension (mast cell-bacteria ratio, 1 : 100) in a microcentrifuge polypropylene tube.

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Adhesion of Microbial Pathogens [no cover, index] by R. Doyle, I. Ofek


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