Supplementary Figure 8: Optical sectioning by DIFF and HiLo.
From: Pan-neuronal calcium imaging with cellular resolution in freely swimming zebrafish
The raw images required for DIFF and HiLo optical sectioning were collected with the following interleaved sequence: I A (DIFF structured image A and HiLo structured image I s), followed by I u (HiLo unstructured image), followed by I B (DIFF structured image B), repeated across an entire imaging session. All images were collected with the same camera gain (21 dB). To ensure all images had the same average brightness, light source was attenuated by two-fold for I u relative to the structured images (to counteract the two-fold increase in the number of “on” DMD pixels). DIFF optical sectioning was performed using I A and I B. HiLo optical sectioning was performed using I s and I u. Top row, DIFF algorithm applied to I A and I B (Methods and Supplementary Notes). Middle and bottom rows, HiLo algorithm48 applied to I s and I u with shot noise correction enabled (middle) or disabled (bottom). Left column, single optically sectioned images. Middle and left columns, temporally averaged images across 5 timepoints (middle) and 20 timepoints (left). To avoid activity related changes in fluorescence, data shown were collected from an anaesthetized 5 dpf larval zebrafish.
