2020/11/23 by Yi Xue, Xue, Yi, David Ren +3 · 2 citations
Biochemistry, Genetics and Molecular Biology · Engineering · #Advanced Fluorescence Microscopy Techniques #Biological Physics (physics.bio-ph) #FOS: Physical sciences #Optical Coherence Tomography Applications #Optics (physics.optics) #Photoacoustic and Ultrasonic Imaging
paper · pdf · doi:10.48550/arxiv.2011.11574
openalex publication_date 2020/11/23 · openalex created_date 2025/10/10 · openalex updated_date 2026/07/28
Fluorescence microscopy is a powerful tool for imaging biological samples with molecular specificity. In contrast, phase microscopy provides label-free measurement of the sample's refractive index (RI), which is an intrinsic optical property that quantitatively relates to cell morphology, mass, and stiffness. Conventional imaging techniques measure either the labeled fluorescence (functional) information or the label-free RI (structural) information, though it may be valuable to have both. For example, biological tissues have heterogeneous RI distributions, causing sample-induced scattering that degrades the fluorescence image quality. When both fluorescence and 3D RI are measured, one can use the RI information to digitally correct multiple-scattering effects in the fluorescence image. Here, we develop a new computational multi-modal imaging method based on epi-mode microscopy that reconstructs both 3D fluorescence and 3D RI from a single dataset. We acquire dozens of fluorescence images, each 'illuminated' by a single fluorophore, then solve an inverse problem with a multiple-scattering forward model. We experimentally demonstrate our method for epi-mode 3D RI imaging and digital correction of multiple-scattering effects in fluorescence images.