Hello. I am working with the dye Sodium Green and I have spent the past two months attempting to get a change in fluorescence by using either the ionophore Gramicidin D, or Veratridine. I did not have any success with Gramicidin D even after trying several concentrations and time duration of treatment. I measured the fluorescence up to 30 minutes after addition of Gramicidin. I was finally able to achieve a change in fluorescence using Veratridine, but the problem is, I don't know if I will be able to perform the dye calibration with this reagent. Do you know if Veratridine functions in the same way as Gramicidin D does? Another problem I have run into is that in order to calibrate for the dye, it is necessary to suspend my cells (lymphoblasts) in solutions of different sodium concentrations. When I do this, my overall result is cell death. The highest concentration of sodium that I use is 145 mM, and for lower concentrations, I substitute for the sodium with potassium. As you may have been able to notice from my message, I have no experience with flow cytometry, but I am determined to accomplish this calibration for Sodium Green. So if you have experience with this dye, or with sodium measurement in general, and if you know any reasons as to why I may be encoutering the problems that I have mentioned previously, I would greatly appreciate your suggestions and comments. Thank you for your attention. David Lima University of Louisville Department of Psychiatry
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