A, representative images of unmixed fluorescence for (left to right) Venus,
Rhodamine 123 (Rh123), unmixing residuals and the merged final image. Spectra for Rh123 and Venus
fluorophores were obtained from Rh123-stained C57BL/6 islets and unstained GLU-Venus islets,
respectively. B, representative unmixed images of Rh123 before photobleaching (0
s), Venus expressing α cells before photobleaching, Rh123 immediately after photobleaching
(117 s) and Rh123 after fluorescence recovery (322 s) in a GLU-Venus islet. C,
percentage fluorescence recovery in α and β cells in a GLU-Venus islet. Data represent
the mean ± s.e.m. over three mice, with each data point averaged over four to 24
cells from three to six islets per mouse. D, fraction of total fluorescence
intensity recovered in α and β cells of a GLU-Venus islet. Data represent the mean
± s.e.m. fraction over three mice for each time-point, with each data point averaged
over four to nine cells from three or four islets per mouse.