FRET imaging microscopy

Brian Herman; Mao Sun; Atsushi Masuda; Hans Gerritsen; Victoria Centonze

doi:10.1117/12.470683

FRET imaging microscopy

Brian Herman, Mao Sun, Atsushi Masuda, Hans Gerritsen, Victoria Centonze

Biomedical Engineering

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

Abstract

Fluorescence Resonance Energy Transfer (FRET) Microscopy has been finding substantial utility in the measurement of a number of biological processes. Most microscopic techniques that have been developed to monitor FRET measure changes in the donor and acceptor emission or fluorescent lifetime of the donor. These include measurements of sensitized emission, acceptor photobleaching and fluorescent lifetime imaging (FLI). However, which of these approaches is the best for a given experimental situation and for use with multiphoton microscopy is not clear. Using mutant GFP FRET caspase-2 substrate targeted to mitochondria, we compare FRET efficiencies measured using sensitized emission, acceptor photobleaching and FLI.

Original language	English (US)
Pages (from-to)	96-105
Number of pages	10
Journal	Proceedings of SPIE - The International Society for Optical Engineering
Volume	4620
DOIs	https://doi.org/10.1117/12.470683
State	Published - Jan 1 2002

Keywords

Acceptor photobleaching and fluorescent lifetime imaging (FLI)
Apoptosis
Fluorescence resonance energy transfer (FRET)

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10.1117/12.470683

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title = "FRET imaging microscopy",

abstract = "Fluorescence Resonance Energy Transfer (FRET) Microscopy has been finding substantial utility in the measurement of a number of biological processes. Most microscopic techniques that have been developed to monitor FRET measure changes in the donor and acceptor emission or fluorescent lifetime of the donor. These include measurements of sensitized emission, acceptor photobleaching and fluorescent lifetime imaging (FLI). However, which of these approaches is the best for a given experimental situation and for use with multiphoton microscopy is not clear. Using mutant GFP FRET caspase-2 substrate targeted to mitochondria, we compare FRET efficiencies measured using sensitized emission, acceptor photobleaching and FLI.",

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AU - Herman, Brian

AU - Sun, Mao

AU - Masuda, Atsushi

AU - Gerritsen, Hans

AU - Centonze, Victoria

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N2 - Fluorescence Resonance Energy Transfer (FRET) Microscopy has been finding substantial utility in the measurement of a number of biological processes. Most microscopic techniques that have been developed to monitor FRET measure changes in the donor and acceptor emission or fluorescent lifetime of the donor. These include measurements of sensitized emission, acceptor photobleaching and fluorescent lifetime imaging (FLI). However, which of these approaches is the best for a given experimental situation and for use with multiphoton microscopy is not clear. Using mutant GFP FRET caspase-2 substrate targeted to mitochondria, we compare FRET efficiencies measured using sensitized emission, acceptor photobleaching and FLI.

AB - Fluorescence Resonance Energy Transfer (FRET) Microscopy has been finding substantial utility in the measurement of a number of biological processes. Most microscopic techniques that have been developed to monitor FRET measure changes in the donor and acceptor emission or fluorescent lifetime of the donor. These include measurements of sensitized emission, acceptor photobleaching and fluorescent lifetime imaging (FLI). However, which of these approaches is the best for a given experimental situation and for use with multiphoton microscopy is not clear. Using mutant GFP FRET caspase-2 substrate targeted to mitochondria, we compare FRET efficiencies measured using sensitized emission, acceptor photobleaching and FLI.

KW - Acceptor photobleaching and fluorescent lifetime imaging (FLI)

KW - Apoptosis

KW - Fluorescence resonance energy transfer (FRET)

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JO - Proceedings of SPIE - The International Society for Optical Engineering

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FRET imaging microscopy

Abstract

Keywords

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