Dear Amanvir!

Please you look at this article :

Article Automated analysis of contractile force and Ca2+ transients ...

Measurement of contractile force and F340-to-F380 ratio.

Hi,

Please go through the following link:

https://www.ionoptix.com/resource/interpolated-numerator-method/

There could be many reasons for not getting a calcium signal:

1) saturation of all the calcium due to overloading with fura2

2) just be sure, add ionomycin or digitonin to see if a transient is seen. Or use EGTA to chelate the calcium signal.

3) as per the attached article, if isobestic measurements are not made frequently (every 30 seconds), there will be error prone results.

Thanks,

Alpana