Cell Digestion Protocol - (Mar/04/2010 )
I'm working at a chemistry lab and our collaborators gave us a sample of a smoker's lung cells for us to characterize the carbon materials in the cell. We initially tried Raman spectroscopy, but (apparently) the lung cells have very high background fluorescence and we couldn't see anything.
So now we're trying to seperate the carbon material from the rest of the cell matter. We tried the freeze/thaw method to lyse the cells. But after centrifuging and taking a Raman of the pellet, we still have very high background fluorescence.
Does anyone have or know where I could find a mild cell digestion protocol that would degrade the cell matter without oxidizing the carbon material? A dilute solution of nitric acid?
This would be great help as I have little background in biology ...
If you the compounds you are looking at are not proteins or bound to protein you could use something like proteinase K in a mild detergent (e.g. 1% SDS) to non-specifically digest the protein and remove the lipids from the solution. These sorts of proceedures are commonly used to extract DNA or RNA from cell lysates.