Figure 1.

Experimental setup of single-objective based multiplexed-multiplane FCS (soSPIM-FCS) measurements. (A) (i) Experimental setup of the soSPIM system. (ii) Closeup of the 45° micromirror angled to reflect the light-sheet beam orthogonal to the detection axis for imaging at two different depths. (B) Principle of multiplane FCS measurements. In multiplane imaging FCS, a sequential time image series is acquired at different z position in a cell and the time autocorrelation performed at every pixel of the plane to achieve multiplane diffusion and concentration maps. (C) Principle of using photoconvertible protein for FCS. (Left) The number of the photoconverted red molecule can be tuned in the observation volume by 405 nm laser excitation. (Right) Dendra2 protein in red form is expected to show increased ACF amplitude. To see this figure in color, go online.