Growing out yeast spheroplasts - (Jun/10/2008 )

Stains which attach to chitin in the cell wall are readily available and could be used to analyze spheroplast populations isolated by simple centrifugation after enzyme treatment/wall disruption . Try Wheat Germ Agglutinin (WGA), calcofluor white or congo red for starters, although there are others. Analysis of the spheroplast fraction with these stains in conjunction with flow cytometry (or even fluorimetry) would give an idea of how many intact cells you have (fluorescence = chitin = cell wall = intact cells). I dont know if the spheroplasts would withstand treatment, but even if they were disrupted, this would still be effective in showing the presence/absence of intact cells. As long as there are no intact cells you should be confident in your isolation method, and even if there are some, you should be able to quantify using this method and use the info to statistically interpret results.