Rapid Frequency-Domain FLIM Spinning Disk Confocal Microscope: Lifetime Resolution, Image Improvement and Wavelet Analysis

被引：0
作者：
Chittanon Buranachai
Daichi Kamiyama
Akira Chiba
Benjamin D. Williams
Robert M. Clegg
机构：
[1] University of Illinois,Center of Biophysics and Computational Biology
[2] Urbana-Champaign,Department of Biology
[3] University of Miami,Cell & Struc Biology
[4] University of Illinois,Department of Physics
[5] Urbana-Champaign,undefined
[6] University of Illinois,undefined
[7] Urbana-Champaign,undefined
来源：
Journal of Fluorescence | 2008年 / 18卷
关键词：
FLIM; FLI; Lifetime imaging; Spinning Disk; Microscope; Polar plot; Wavelet; Morphology; Background subtraction; Denoising;
D O I：
暂无
中图分类号：
学科分类号：
摘要：
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.
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页码：929 / 942
页数：13
相关论文
共 122 条
  • [1] Gadella TWJ(1993)Fluorescence lifetime imaging microscopy (FLIM): Spatial resolution of microstructures on the nanosecond time scale Biophys Chemist 48 221-239
  • [2] Jovin TM(1995)Possibility of simultaneously measuring low and high calcium concentrations using Fura-2 and lifetime-based sensing Cell Calcium 18 64-75
  • [3] Clegg RM(2003)Imaging fluorescence lifetime modulation of a ruthenium-based dye in living cells: the potential for oxygen sensing J Phys D: Appl Phys 36 1689-1695
  • [4] Szmacinski H(2003)Fluorescence lifetime-resolved imaging: measuring lifetimes in an image Methods Enzymol 360 509-542
  • [5] Lakowicz JR(2005)Fluorescence Lifetime Imaging Microscopy (FLIM) Adv Biochem Eng Biotechnol 95 143-175
  • [6] Zhong W(2005)Time-resolved fluorescence microscopy Photochem Photobiol Sci 4 13-22
  • [7] Urayama P(1991)Time resolved imaging microscopy. Phosphorescence and delayed fluorescence imaging Biophys J 60 1374-1387
  • [8] Mycek MA(1991)Use of time-gated fluorescence imaging for diagnosis in biomedicine J Photochem Photobiol, B Biol 12 109-113
  • [9] Clegg RM(2002)Time-resolved fluorescence imaging in biology and medicine J Phys, D, Appl Phys 35 R61-R76
  • [10] Holub O(2003)Fluorescence lifetime imaging for the two-photon microscope: time-domain and frequency-domain methods J of Biomedical Optics 8 381-390
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