Control, MIC level and MFC level of AuNPs treated C. albicans cell were harvested after 6 h expose time. The cells were washed with PBS, incubated with 5 mg/L rhodamine 123 (Rho123,Sigma,USA) at 37 0C for 30 min in the dark, and then washed twice with PBS. The cell fluorescence intensity at 535 nm was quanyified with a BD FACSCanto II system flow cytometer (Bectone Dickinson,USA). A total of 50,000 events were evaluated per experiment (n= 2), and the cellular debris was omitted from the analysis.
Dear Sajith Dananjaya,
In damaged cells there is an enhanced permeation of the dye inside the cell. This results in binding to a variety of species thus increasing the fluorescence intensity.
Please note, that NOT only rhodamine 123 does that, many other fluorescence dyes do the same.
For more information regarding this topic, please use the following link:
http://www.sciencedirect.com/science/article/pii/S0011224014001205
http://www.scielo.br/pdf/fb/v31n4/04.pdf
https://www.dojindo.com/Protocol/CellStaining_Protocol.pdf
Hoping this will be helpful,
Rafik
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