title: Quantitative comparison between full-spectrum and filter-based imaging in hyperspectral fluorescence microscopy.
abstract: We implement a filterless illumination scheme on a hyperspectral fluorescence microscope to achieve full-range spectral imaging. The microscope employs polarisation filtering, spatial filtering and spectral unmixing filtering to replace the role of traditional filters. Quantitative comparisons between full-spectrum and filter-based microscopy are provided in the context of signal dynamic range and accuracy of measured fluorophores' emission spectra. To show potential applications, a five-colour cell immunofluorescence imaging experiment is theoretically simulated. Simulation results indicate that the use of proposed full-spectrum imaging technique may result in three times improvement in signal dynamic range compared to that can be achieved in the filter-based imaging.
authors.affiliation: Department of Bioengineering, Rice University, 6100 Main Street, Houston, TX 77005, USA.
authors.forename: L
authors.initials: L
authors.lastname: Gao
authors.forename: N
authors.initials: N
authors.lastname: Hagen
authors.forename: T S
authors.initials: TS
authors.lastname: Tkaczyk
mesh_terms: D000818:Animals; D002417:Cattle; D042783:Endothelial Cells / Q000648:ultrastructure*; D005374:Filtration / Q000295:instrumentation*; D005456:Fluorescent Dyes / Q000378:metabolism*; D021621:Imaging, Three-Dimensional / Q000379:methods*; D008856:Microscopy, Fluorescence / Q000295:instrumentation / Q000379:methods*; D011651:Pulmonary Artery / Q000166:cytology*; D013050:Spectrometry, Fluorescence / Q000295:instrumentation / Q000379:methods
chemical_list: D005456:Fluorescent Dyes