Journal of Fluorescence

, Volume 18, Issue 5, pp 929–942

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
Original Paper

DOI: 10.1007/s10895-008-0332-3

Cite this article as:
Buranachai, C., Kamiyama, D., Chiba, A. et al. J Fluoresc (2008) 18: 929. doi:10.1007/s10895-008-0332-3

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.

Keywords

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

Copyright information

© Springer Science+Business Media, LLC 2008

Authors and Affiliations

  • Chittanon Buranachai
    • 1
  • Daichi Kamiyama
    • 2
  • Akira Chiba
    • 2
  • Benjamin D. Williams
    • 3
  • Robert M. Clegg
    • 4
  1. 1.Center of Biophysics and Computational BiologyUniversity of Illinois, Urbana-ChampaignUrbanaUSA
  2. 2.Department of BiologyUniversity of MiamiCoral GablesUSA
  3. 3.Cell & Struc BiologyUniversity of Illinois, Urbana-ChampaignUrbanaUSA
  4. 4.Department of PhysicsUniversity of Illinois, Urbana-ChampaignUrbanaUSA

Personalised recommendations