Quantitative comparison between full-spectrum and filter-based imaging in hyperspectral fluorescence microscopy

L. Gao, N. Hagen, T. S. Tkaczyk

Research output: Contribution to journalArticlepeer-review

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 languageEnglish (US)
Pages (from-to)113-123
Number of pages11
JournalJournal of Microscopy
Volume246
Issue number2
DOIs
StatePublished - May 2012
Externally publishedYes

Keywords

  • Filterless illumination
  • Hyperspectral fluorescence microscopy
  • Spectroscopy

ASJC Scopus subject areas

  • Pathology and Forensic Medicine
  • Histology

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