Figure 4

AutoAO and FOCALS. (a) FLIM and OCM images of NE-4C neuroepithelial cells under different magnitudes of OAs applied to the DM. The first column shows the zoomed-in lifetime images corresponding to the white dashed box in the second column; the third and fourth columns show the OCM images and the DM patterns, respectively. (b) Histogram of the NAD(P)H fluorescence lifetime for OA magnitudes −0.4 (blue), 0.4 (gray), and 0 (orange). (c–e) Plot of (c) mean NAD(P)H fluorescence intensity per frame, (d) mean lifetime (in ps), and (e) number of pixels with a lifetime of 1500 ps or longer against OA magnitude. (f) FLIM, OCM, and PS-OCM (polarization ratio) images of a rat kidney 5 µm below the surface, acquired with a flat pattern on the DM, and 45 µm below the surface before and after aberration correction with AO, with three different zoomed-in regions of interest highlighted by the blue, orange, and cyan boxes. (g) Histogram of the NAD(P)H fluorescence lifetime for the kidney 45 µm below the surface before (blue) and after (orange) OA correction using AutoAO. (h) The DM pattern generated by the AutoAO algorithm. (i) FLIM, OCM, polarization ratio, and SHG images before and after AutoAO correction of a mouse muscle 40 µm below the surface. The orange dots correspond to pixels that have a lifetime of 1500 ps or longer. The white arrows indicate the scattering regions in the OCM image around which the orange dots are clustered. The magenta arrows highlight the regions where the fibrous structures are apparent in the SHG images and the polarization ratio is biased towards a particular state. (j) Histogram of the NAD(P)H fluorescence lifetime before (blue) and after (orange) OA correction using AutoAO. (k) The DM pattern generated by the AutoAO algorithm.