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

被引：35

作者：

Buranachai, Chittanon ^{[2
]}

Kamiyama, Daichi ^{[3
]}

Chiba, Akira ^{[3
]}

Williams, Benjamin D.

Clegg, Robert M. ^{[1
]}

机构：

[1] Univ Illinois, Dept Phys, Loomis Lab, Urbana, IL 61801 USA

[2] Univ Illinois, Ctr Biophys & Computat Biol, Loomis Lab, Urbana, IL 61801 USA

[3] Univ Miami, Dept Biol, Coral Gables, FL 33124 USA

来源：

JOURNAL OF FLUORESCENCE | 2008年 / 18卷 / 05期

关键词：

FLIM; FLI; lifetime imaging; spinning disk; microscope; polar plot; wavelet; morphology; background subtraction; denoising;

D O I：

10.1007/s10895-008-0332-3

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

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.

引用

页码：929 / 942

页数：14

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