Holographic Microscopy for Cell Imaging

The holographic microscopy setup was similar to that used previously (Singh et al., 2018 (link); Kühn et al., 2018 (link); Thornton et al., 2020 (link)), with a few modifications. The samples were imaged on a Nikon Eclipse E600 upright microscope. The illumination source was a single-mode fibre-coupled laser diode with peak emission at 642 nm. The end of the fibre was mounted below the specimen stage using a custom adaptor and delivered a total of 15 mW of optical power to the sample. A Mikrotron MC-1362 monochrome camera was used to acquire videos of 3000 frames, at a frame rate of 50 Hz and with an exposure time of 100 μs. A 10× magnification bright-field lens with numerical aperture of 0.3 was used to acquire data at a video resolution of 1024 × 1024 pixels², corresponding to a field of view measuring 1.44 × 1.44 mm². The raw videos were saved as uncompressed, 8-bit AVI files. Three biological replicates of each condition were prepared, with five technical replicates from each of these.