Advise on cellular fluorescent assay - (Feb/24/2012 )

Hi all

Just wondering if anyone would have a thought on the following experiment.

We are segregating 2 cell types from a mixed culture using a fluorescent marker and an optical tweezers. Essentially, red-flu cells into one pot, and green-flu cells into another.

Can anyone recommend an assay that would then confirm the segregation based on the internal metabolism of the cells? I would rather not simply use a second fluorescent epitope marker as this would invalidate the need for the separation in the first place.

I was hoping to use a metabolic marker (for instance, the concentration of NADH and NADPH) between two cell lines, but I'm having difficulty finding 2 cell lines in which there is a confirmed measurable difference.

Metabolics isn't my area of expertise, and the assay itself is not very important as it is the separation technique that we are looking to highlight rather than intricate biology. Can anyone suggest a metabolic (preferably fluorescent) assay that would demonstrate a difference in activity between 2 cell lines?

Many thanks all

Hej,

I used to use Alamar Blue dye (Resazurin) to check the metabolic activity in cells. It worked good for me.

What are the two cell populations you are trying to separate? This is important to know to check if they have different metabolism.