High-speed confocal fluorescence lifetime imaging microscopy (FLIM) with the analog mean delay (AMD) method

Youngjae Won; Sucbei Moon; Wenzhong Yang; Donguk Kim; Won Taek Han; Dug Young Kim

doi:10.1364/OE.19.003396

High-speed confocal fluorescence lifetime imaging microscopy (FLIM) with the analog mean delay (AMD) method

Youngjae Won, Sucbei Moon, Wenzhong Yang, Donguk Kim, Won Taek Han, Dug Young Kim

Department of Physics

Research output: Contribution to journal › Article › peer-review

44 Citations (Scopus)

Abstract

We demonstrate a high-speed confocal fluorescence lifetime imaging microscopy (FLIM) whose accuracy and photon economy are as good as that of a time-correlated single photon counting (TCSPC). It is based on a new lifetime determination scheme, the analog mean delay (AMD) method. Due to the technical advantages of multiple fluorescence photon detection capability, accurate lifetime determination scheme and high photon detection efficiency, the AMD method can be the most effective method for high-speed confocal FLIM. The feasibility of real-time confocal FLIM with the AMD method has been demonstrated by observing the dynamic reaction of calcium channels in a RBL-2H3 cell with respect to 4αPDD stimulus. We have achieved the photon detection rate of 125 times faster than a conventional TCSPC based system in this experiment.

Original language	English
Pages (from-to)	3396-3405
Number of pages	10
Journal	Optics Express
Volume	19
Issue number	4
DOIs	https://doi.org/10.1364/OE.19.003396
Publication status	Published - 2011 Feb 14

All Science Journal Classification (ASJC) codes

Atomic and Molecular Physics, and Optics

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abstract = "We demonstrate a high-speed confocal fluorescence lifetime imaging microscopy (FLIM) whose accuracy and photon economy are as good as that of a time-correlated single photon counting (TCSPC). It is based on a new lifetime determination scheme, the analog mean delay (AMD) method. Due to the technical advantages of multiple fluorescence photon detection capability, accurate lifetime determination scheme and high photon detection efficiency, the AMD method can be the most effective method for high-speed confocal FLIM. The feasibility of real-time confocal FLIM with the AMD method has been demonstrated by observing the dynamic reaction of calcium channels in a RBL-2H3 cell with respect to 4αPDD stimulus. We have achieved the photon detection rate of 125 times faster than a conventional TCSPC based system in this experiment.",

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AU - Won, Youngjae

AU - Moon, Sucbei

AU - Yang, Wenzhong

AU - Kim, Donguk

AU - Han, Won Taek

AU - Kim, Dug Young

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AB - We demonstrate a high-speed confocal fluorescence lifetime imaging microscopy (FLIM) whose accuracy and photon economy are as good as that of a time-correlated single photon counting (TCSPC). It is based on a new lifetime determination scheme, the analog mean delay (AMD) method. Due to the technical advantages of multiple fluorescence photon detection capability, accurate lifetime determination scheme and high photon detection efficiency, the AMD method can be the most effective method for high-speed confocal FLIM. The feasibility of real-time confocal FLIM with the AMD method has been demonstrated by observing the dynamic reaction of calcium channels in a RBL-2H3 cell with respect to 4αPDD stimulus. We have achieved the photon detection rate of 125 times faster than a conventional TCSPC based system in this experiment.

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High-speed confocal fluorescence lifetime imaging microscopy (FLIM) with the analog mean delay (AMD) method

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