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Flow cytometric analysis of S. epidermidis and its biofilm components

JH Jian Hu
NJ Ning Zhang, Jr.
LL Lifang Li
NS Ning Zhang, Sr.
YM Yanfen Ma
CZ Chedong Zhao
QW Qian Wu
YL Ying Li
NH Nianan He
XW Xiaoqin Wang
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Overnight bacterial culture was centrifuged to collect the planktonic S. epidermidis. After washing thrice with PBS, the collected bacteria were fully suspended and diluted to 0.5 McIntosh turbidity unit in fluorescent staining solution containing 0.67 μmol/L 5-CFDA and 0.33 mmol/L CTC (in normal saline) before flow cytometric analysis (Cantoll, Becton Dickinson and Company, New Jersey, USA).

For obtaining biofilm cells, overnight cultured biofilm was washed with PBS for removal of non-adherent bacteria. After ultrasonic or UTMD treatment, the biofilm was fully scraped and cells were collected after centrifugation. After washing thrice with PBS, the collected biofilm components were fully suspended and diluted to 0.5 McIntosh turbidity unit in fluorescent staining solution containing 5-CFDA and CTC before flow cytometric analysis.

Copyright and License information: The Author(s) ©2017
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