 |
 |
 |
 |
>Raymond, CHANG asked about fluo-3 and calcium measurements.
We try to useFluo-3. In our studies, the cells were incubated in a temp- and
pH controlled chamber and we hope to measure the Ca2+ up to 90 mins, i.e. to
take the cells into the flow every 5 minutes for measurement. We wonder
the dye is suitable for such long time incubation at 37C. Is there anyone
who has such experience?
I have used Fluo-3 to measure Ca2+ in thymocytes over 80 minutes. This method
involved pre-incubating the cells for 10 mins at 2e7 cell/mL with dye at
6ug/mL of cells at 37oC. The sample was then diluted to 1e6 cell/mL and run
collecting data at 5 sec intervals. It worked very well. Larger cells may
need a higher concentration of fluo-3. I used thapsigargin at 10nM or 100nM
as a positive control each day to make sure the set up was still OK. It should
yield a large increase in intracellular calcium within a few minutes.
Jo Beaver
|
|
|
|