Multimodal Spectral Imaging of Cells Using a Transmission Diffraction Grating on a Light Microscope

作者：Isailovic Dragan^*; Xu Yang; Copus Tyler; Saraswat Suraj; Nauli Surya M

来源：Applied Spectroscopy, 2011, 65(6): 575-583.

DOI：10.1366/10-06104

• Spectral imaging
• Single cell
• Transmission diffraction grating
• Light microscopy
• Spectroscopy
• Fluorescence
• Brightfield
• Darkfield
• Differential interference contrast
• DIC

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.

• 出版日期2011-6