Heat and Chemical Treatments Affect the Viability, Morphology, and Physiology of Staphylococcus aureus and Its Subsequent Antibody Labeling for Flow Cytometric Analysis

Author:

Kennedy Deirdre1,Cronin Ultan P.2ORCID,Piterina Anna3,Wilkinson Martin G.2

Affiliation:

1. Food Chemistry and Technology Department, Teagasc Food Research Centre, Moorepark, Fermoy, County Cork, Ireland

2. Department of Biological Sciences, University of Limerick, Limerick, Ireland

3. Dairy Processing Technology Centre, University of Limerick, Limerick, Ireland

Abstract

FCM data indicated that cells conventionally considered to be dead and which would not give rise to CFU in a plate count assay, e.g., cells heated to 80°C, were labeled by antibody staining. This finding suggests that without the inclusion of a live/dead discriminating dye, these cells would be erroneously detected as viable within an FCM assay. Reductions in antibody staining due to physicochemical treatment were strain related, reflecting the complexity of the phenomenon under study and illustrating that substantial validation of any new antibody detection-based method, including physiological staining and cell sorting, should be undertaken. Researchers should be aware of physicochemical treatments causing false-negative results: in this study, freeze-thawing severely reduced antibody binding without affecting the viability of a substantial percentage of cells. Scanning electron microscopy carried out on treated cells revealed a range of morphological changes resulting from physicochemical treatments which may have hindered antibody binding.

Funder

Department of Agriculture, Food and the Marine

Publisher

American Society for Microbiology

Subject

Ecology,Applied Microbiology and Biotechnology,Food Science,Biotechnology

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