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

doi:10.1007/s10895-008-0332-3

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 journal › Article › peer-review

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 language	English (US)
Pages (from-to)	929-942
Number of pages	14
Journal	Journal of Fluorescence
Volume	18
Issue number	5
DOIs	https://doi.org/10.1007/s10895-008-0332-3
State	Published - Sep 2008

Keywords

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

ASJC Scopus subject areas

Biochemistry
Spectroscopy
Clinical Biochemistry

Online availability

10.1007/s10895-008-0332-3

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

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

author = "Chittanon Buranachai and Daichi Kamiyama and Akira Chiba and Williams, {Benjamin D.} and Clegg, {Robert M.}",

note = "Funding Information: Acknowledgments We thank Glen Redford for his valuable contributions to the non-confocal version of the frequency domain full field FLIM, and his original work on the polar plot. We appreciate discussions with Bryan Spring about wavelets. The work presented here has been partially supported by the NIH grant (PHS 5 P41 RRO3155) and by start-up funds from the UIUC Physics Department (RMC).",

year = "2008",

month = sep,

doi = "10.1007/s10895-008-0332-3",

language = "English (US)",

volume = "18",

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journal = "Journal of Fluorescence",

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T2 - Lifetime resolution, image improvement and wavelet analysis

AU - Buranachai, Chittanon

AU - Kamiyama, Daichi

AU - Chiba, Akira

AU - Williams, Benjamin D.

AU - Clegg, Robert M.

N1 - Funding Information: Acknowledgments We thank Glen Redford for his valuable contributions to the non-confocal version of the frequency domain full field FLIM, and his original work on the polar plot. We appreciate discussions with Bryan Spring about wavelets. The work presented here has been partially supported by the NIH grant (PHS 5 P41 RRO3155) and by start-up funds from the UIUC Physics Department (RMC).

PY - 2008/9

Y1 - 2008/9

N2 - 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.

AB - 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.

KW - Background subtraction

KW - Denoising

KW - FLI

KW - FLIM

KW - Lifetime imaging

KW - Microscope

KW - Morphology

KW - Polar plot

KW - Spinning Disk

KW - Wavelet

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

Abstract

Keywords

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