An EGFP-chimera of a GPI-anchored was expressed in HEK293 cells. The distribution of the protein in a membrane of living cells was imaged using Alba FLIM™. Cells were plated on a glass-bottom 35 mm Petri dish, kept in medium (HEPES buffer) and imaged at room temperature. Images were acquired using Alba FLIM™ equipped with the piezo-controlled stage using single photon excitation.
The Z-stack was acquired with single photon excitation at 488 nm. Emission was passed trough a pinhole of 50 μm. Images of 256x256 pixels were acquired with an integration time of 1.0 ms.
Courtesy of Drs. V. Caiolfa, N. Sidenius and F. Blasi at the Scientific Institute San Raffaele, Milano (Italy).