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Fig 1.

(a) In-focus subimages cropped from a 9 plane z stack acquired from the camera resulting from deploying a Pixelflipper algorithm optimized SLM display pattern. The emission light is unevenly distributed in the subimages, with the central zeroth order subimage receiving most of the emission light, and (b) zoomed-in view of a 64 × 64 pixels2 region of the SLM displayed grating pattern which gives the multifocal subimages in (a).

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Fig 2.

Block diagram illustrating in situ iterative algorithm implementation.

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Fig 3.

(a) In-focus subimages cropped from a 9 plane z stack acquired resulting from deploying our in situ iteratively optimized SLM display pattern. The emission light striking the camera is more evenly distributed among the subimages compared to Fig 1(a), and (b) zoomed-in view of a 64 × 64 pixels2 region of the SLM displayed grating pattern which gives the images in (a).

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Fig 4.

(a) Boxplots of output M values resulting from grating patterns optimized using Pixflipper (500 iterations, Randomized (500 iterations), IFTA (500 iterations) and our in situ iterative algorithm (12 iterations), (b) image resulting from Pixelflipper algorithm generated pattern having M = 0.062, (c) image resulting from IFTA algorithm generated pattern (M = 0.211), and (d) image resulting from in-situ iterative algorithm generated pattern (M = 0.644).

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Fig 5.

Multifocal images of different regions of a microtubule stained Hela Kyoto Cells fixed sample with (a) Δz = 0.50 μm, and (b) Δz = 1.00 μm.

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