Extended Data Fig. 4: Signal-to-noise in vitro analysis of the PTEN sensor.
From: Genetically encoded biosensor for fluorescence lifetime imaging of PTEN dynamics in the intact brain
a-b Representative pseudo-colored images of fluorescent lifetime summed for different frame duration, for G-PTEN before and after 3 hr of 50 μM TBB application. Representative images represent experiments repeated independently at least 3 times. Scale bar; 50 μm. c-d Standard deviation of signal-to-noise of mEGFP-PTEN-sREACh before and after TBB, as a function of number of photons from each cell. Each point represents individual cells for frame-to-frame (left) or individual experiments over one field of view for cell-to-cell (right) variation. The binding fraction’s theoretical change caused by shot noise is represented by the black curve (see methods for equation). e-f Standard deviation of signal-to-noise of G-PTEN before and after TBB, as a function of the number of photons from each cell. Each point represents individual cells for frame-to-frame (left) or individual experiments over one field of view for cell-to-cell (right). The binding fraction’s theoretical change caused by shot noise is represented by the black curve.
