Live dead bac light bacterialviability kits

The viability of the Yersinia strain during mineralization was monitored in MASE I medium using the Most Probable Number (MPN) method. For that, the cell density of the preculture used for mineralization was determined by cell counting with a Thoma chamber and the positive growth of serial dilutions (from non-diluted to dilution 10⁻⁸) was checked by optical microscopy.
Live/dead staining was also performed to distinguish cells with possible disrupted membranes from cells with probable intact membranes using LIVE/DEAD^® Bac Light^TM BacterialViability Kits (ThermoFisher Scientific), according to manufacturer’s instructions. Stained cells were observed by epifluorescence microscopy with a Zeiss Axiovert 200 microscope, equipped with an Hg-lamp. Acquisitions were made with the AxioVision software.

The P. aeruginosa strain (ATCC 9027) was cultured at 37°C until the bacterial cells reached the logarithmic phase, and then the bacterial cells were harvested and washed twice using the NB. The pellet was resuspended to ~10⁶ CFU·ml⁻¹ in the same buffer, after which the prepared bacteria were treated as mentioned above. After the bacteria were treated with all the antibacterial agents, all the bacteria were harvested and stained with SYTO 9 and propidium iodide (PI) in the ratio of 1:1 from the LIVE/DEAD® BacLight^TM Bacterial Viability Kits (Thermo Fisher Scientific). After mixing all the mixture thoroughly, the CytoFLEX flow cytometry (Beckman Coulter, California, USA) was used to test the cell membrane integrity and cell viability of the bacteria.