Although the contrast generated in transmitted light microscopy is due to the elastic scattering of light, multiple scattering scrambles the image and reduces overall visibility. To image both thin and thick samples, we turn to gradient light interference microscopy (GLIM) to simultaneously measure morphological parameters such as cell mass, volume, and surfaces as they change through time. Because GLIM combines multiple intensity images corresponding to controlled phase offsets between laterally sheared beams, incoherent contributions from multiple scattering are implicitly cancelled during the phase reconstruction procedure. As the interfering beams traverse near identical paths, they remain comparable in power and interfere with optimal contrast. This key property lets us obtain tomographic parameters from wide field z-scans after simple numerical processing. Here we show our results on reconstructing tomograms of bovine embryos, characterizing the time-lapse growth of HeLa cells in 3D, and preliminary results on imaging much larger specimen such as brain slices.
Mikhail E. Kandel, Ghazal N. Kouzehgarani, Tan H. Ngyuen, Martha U. Gillette, and Gabriel Popescu, "Label-free tomographic reconstruction of optically thick structures using GLIM (Conference Presentation)," Proc. SPIE 10074, Quantitative Phase Imaging III, 1007403 (Presented at SPIE BiOS: January 29, 2017; Published: 24 April 2017); https://doi.org/10.1117/12.2257046.5380600131001.
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