Rapid frequency-domain flim spinning disk confocal microscope: Lifetime resolution, image improvement and wavelet analysis

Chittanon Buranachai, Daichi Kamiyama, Akira Chiba, Benjamin D. Williams, Robert M. Clegg

Research output: Contribution to journalArticle

Abstract

A spinning disk confocal attachment is added to a full-field real-time frequency-domain fluorescence lifetime-resolved imaging microscope (FLIM). This provides confocal 3-D imaging while retaining all the characteristics of the normal 2-D FLIM. The spinning disk arrangement allows us to retain the speed of the normal 2-D full field FLIM while gaining true 3-D resolution. We also introduce the use of wavelet image transformations into the FLIM analysis. Wavelets prove useful for selecting objects according to their morphology, denoising and background subtraction. The performance of the instrument and the analysis routines are tested with quantitative physical samples and examples are presented with complex biological samples.

Original languageEnglish (US)
Pages (from-to)929-942
Number of pages14
JournalJournal of Fluorescence
Volume18
Issue number5
DOIs
StatePublished - Sep 1 2008

Keywords

  • Background subtraction
  • Denoising
  • FLI
  • FLIM
  • Lifetime imaging
  • Microscope
  • Morphology
  • Polar plot
  • Spinning Disk
  • Wavelet

ASJC Scopus subject areas

  • Biochemistry
  • Clinical Psychology
  • Social Sciences (miscellaneous)
  • Sociology and Political Science
  • Spectroscopy
  • Clinical Biochemistry
  • Law

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