Fig. 3. FLIM images of HK cells labelled with ER flipper 4 before (a, c) and after (b, d) hyper-osmotic shock, showing average lifetimes τav (a, b) and τ1 (c, d) from triexponential reconvolution; scale bars = 10 μm. (e) Distribution of the photon counts associated with the τ1 component of 4 in HK cells after triexponential reconvolution of FLIM images before (c, τ1i) and after (d, τ1h) hyper-osmotic shock, showing decreasing lifetimes for τ1 (4d). (f) The dehydration factor dhi defined as total integrated photon counts for τ1 (Στ1) divided by Στ2 (i.e., dhi = area Στ1i/area Στ2i) for 4 in strongly hydrated ER (dhi < 2, turquoise) and 1 in weakly hydrated plasma membrane (dhi > 6, purple) of HK Kyoto cells under iso-osmotic conditions.
