Refractive-index (RI) is an inherent optical property of materials that can provide important biochemical and biophysical information about a biological sample. Optical-diffraction-tomography (ODT) is a current standard to obtain quantitative three-dimensional RI distributions, by measuring optical fields diffracted from the sample by rotated illumination beams. This method for ODT also synthetically enlarges the microscope’s lateral spatial-frequency support, and thus images the RI distribution with lateral resolution beyond the microscope's coherent diffraction limit. Fluorescence microscopy offers a complementary set of biological insights by offering imaging capabilities with molecular specificity. Analogous to ODT, super-resolution fluorescence techniques can offer these insights at spatial resolutions beyond the microscope's incoherent diffraction limit. Unfortunately, such super-resolution techniques are generally incompatible with ODT and a generalized sub-diffraction technique has been difficult to find, which hinders a cohesive, high-resolution, multimodal analysis of biological samples. We experimentally introduce, for the first time to our knowledge, a novel, high resolution, optical system that uses structured illumination (SI) to enable 3D sub-diffraction resolution imaging for both fluorescence and RI. We demonstrate sub-diffraction resolution, multimodal SI imaging of HT29 and MCF7 cells fluorescently stained for F-actin, such that the 3D RI and fluorescent distributions may offer unique, but complementary, insights into the biological samples.
Shwetadwip Chowdhury and Joseph A. Izatt, "Structured illumination for 3D subdiffraction reconstruction of refractive-index and fluorescence (Conference Presentation)," Proc. SPIE 10074, Quantitative Phase Imaging III, 100740N (Presented at SPIE BiOS: January 30, 2017; Published: 24 April 2017); https://doi.org/10.1117/12.2253408.5380600132001.
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