What are the fluorescence excitation/emission maxima for the SYTO 9 dye and propidium iodide in the LIVE/DEAD FungaLight Yeast Viability Kit, for flow cytometry?
How do I prepare dead cell controls for LIVE/DEAD cell viability assays?
There are two easy options. One is to heat-inactivate the cells by placing at 60 degrees C for 20 minutes. The second is to subject the cells to 70% ethanol. Alcohol-fixed cells can be stored indefinitely in the freezer until use, potentially up to several years.
Centrifuge cells, pellet, and remove supernatant.
Fix cells: Add 10 mL ice cold 70% ETOH to a 15 mL tube containing the cell pellet, adding dropwise at first while vortexing, mix well.
Store in freezer until use.
When ready to use, wash twice and resuspend in buffer of choice.
Quantification of Saccharomyces cerevisiae viability using BacLight.
Authors:Zhang T, Fang HH
Journal:Biotechnol Lett
PubMed ID:15269525
Yeast viability can be accurately quantified using BacLight, a kit which so far has been used only for bacterial analysis. Upon staining, viable cells can be differentiated from non-viable ones by either confocal laser scanning microscopy (CLSM), epifluorescence microscopy, or flow cytometry. Using Saccharomyces cerevisiae as a model, viabilities quantified ... More