Multimodal Spectral Imaging of Cells Using a Transmission Diffraction Grating on a Light Microscope
A multimodal methodology for spectral imaging of cells is presented. The spectral imaging setup uses a transmission diffraction grating on a light microscope to concurrently record spectral images of cells and cellular organelles by fluorescence, darkfield, brightfield, and differential
interference contrast (DIC) spectral microscopy. Initially, the setup was applied for fluorescence spectral imaging of yeast and mammalian cells labeled with multiple fluorophores. Fluorescence signals originating from fluorescently labeled biomolecules in cells were collected through triple
or single filter cubes, separated by the grating, and imaged using a charge-coupled device (CCD) camera. Cellular components such as nuclei, cytoskeleton, and mitochondria were spatially separated by the fluorescence spectra of the fluorophores present in them, providing detailed multi-colored
spectral images of cells. Additionally, the grating-based spectral microscope enabled measurement of scattering and absorption spectra of unlabeled cells and stained tissue sections using darkfield and brightfield or DIC spectral microscopy, respectively. The presented spectral imaging methodology
provides a readily affordable approach for multimodal spectral characterization of biological cells and other specimens.
DIFFERENTIAL INTERFERENCE CONTRAST;
INDEX HEADINGS: SPECTRAL IMAGING;
TRANSMISSION DIFFRACTION GRATING
Document Type: Research Article
Department of Chemistry, University of Toledo, Toledo, Ohio 43606, USA. Dragan.Isailovic@utoledo.edu
Department of Chemistry, University of Toledo, Toledo, Ohio 43606, USA
Department of Chemistry, University of Toledo, Toledo, Ohio 43606, USA; Findlay High School, Findlay, OH 45840, USA
Department of Pharmacology, University of Toledo, Toledo, Ohio 43606, USA
Publication date: June 1, 2011
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