Journal papers
Yi Xue; David Ren; Laura Waller
Three-dimensional bi-functional refractive index and fluorescence microscopy (BRIEF) Journal Article
In: Biomed. Opt. Express, vol. 13, no. 11, pp. 5900–5908, 2022.
Abstract | Links | BibTeX | Tags: Digital imaging; Fluorescence microscopy; Image quality; Imaging techniques; Optical imaging; Three dimensional imaging
@article{Xue:22,
title = {Three-dimensional bi-functional refractive index and fluorescence microscopy (BRIEF)},
author = {Yi Xue and David Ren and Laura Waller},
url = {https://opg.optica.org/boe/abstract.cfm?URI=boe-13-11-5900},
doi = {10.1364/BOE.456621},
year = {2022},
date = {2022-11-01},
journal = {Biomed. Opt. Express},
volume = {13},
number = {11},
pages = {5900--5908},
publisher = {Optica Publishing Group},
abstract = {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.},
keywords = {Digital imaging; Fluorescence microscopy; Image quality; Imaging techniques; Optical imaging; Three dimensional imaging},
pubstate = {published},
tppubtype = {article}
}
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.