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.
Original language | English (US) |
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Pages (from-to) | 113-123 |
Number of pages | 11 |
Journal | Journal of Microscopy |
Volume | 246 |
Issue number | 2 |
DOIs | |
State | Published - May 2012 |
Externally published | Yes |
Keywords
- Filterless illumination
- Hyperspectral fluorescence microscopy
- Spectroscopy
ASJC Scopus subject areas
- Pathology and Forensic Medicine
- Histology