Alpha Hemolysin Induces an Increase of Erythrocytes Calcium: A FLIM 2-Photon Phasor Analysis Approach
Figure 2
Separation of auto fluorescence from CaG-1 signal in RRBC.
Phasor representation of FLIM images acquired on RRBC incubated with and without CaG-1. For comparison purposes, the same power and the same integration time were used to obtain the images from both samples. The auto fluorescence corresponds to the signal obtained from the RRBC without CaG-1, Data were analyzed calculating the phasors at the basic laser frequency (80 MHz) (A) and at the second harmonic of it (160 MHz) (B). Using the second harmonic the cluster of phasors from the auto fluorescence can be separated from the CaG-1 phasors and the concentration of Ca+2 inside the RRBC can be determined.