Removal of background signals from fluorescence thermometry measurements in PDMS microchannels using fluorescence lifetime imaging

Tom Robinson; Yolanda Schaerli; Robert Wootton; Florian Hollfelder; Christopher Dunsby; Geoff Baldwin; Mark Neil; Paul French; Andrew Demello

doi:10.1039/b913293g

Removal of background signals from fluorescence thermometry measurements in PDMS microchannels using fluorescence lifetime imaging

Tom Robinson, Yolanda Schaerli, Robert Wootton, Florian Hollfelder, Christopher Dunsby, Geoff Baldwin, Mark Neil, Paul French^*, Andrew Demello

^*Corresponding author for this work

School of Engineering

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

Abstract

We report a method for removing unwanted contributions to fluorescence signals from dyes absorbed in polydimethylsiloxane (PDMS) using fluorescence lifetime imaging microscopy (FLIM). By analysing experimental fluorescence decays using a bi-exponential decay model, we are able to discriminate between emission originating from dye molecules in free solution and those absorbed within the PDMS substrate. Simple image processing allows the unwanted background signal to be removed and thus enables a more accurate assessment of temperature. The efficacy of the approach is demonstrated by measuring temperature changes within a droplet-based PCR device.

Original language	English
Pages (from-to)	3437-3441
Number of pages	5
Journal	Lab on a chip
Volume	9
Issue number	23
DOIs	https://doi.org/10.1039/b913293g
Publication status	Published - 24 Sept 2009

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10.1039/b913293g

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abstract = "We report a method for removing unwanted contributions to fluorescence signals from dyes absorbed in polydimethylsiloxane (PDMS) using fluorescence lifetime imaging microscopy (FLIM). By analysing experimental fluorescence decays using a bi-exponential decay model, we are able to discriminate between emission originating from dye molecules in free solution and those absorbed within the PDMS substrate. Simple image processing allows the unwanted background signal to be removed and thus enables a more accurate assessment of temperature. The efficacy of the approach is demonstrated by measuring temperature changes within a droplet-based PCR device.",

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AU - Robinson, Tom

AU - Schaerli, Yolanda

AU - Wootton, Robert

AU - Hollfelder, Florian

AU - Dunsby, Christopher

AU - Baldwin, Geoff

AU - Neil, Mark

AU - French, Paul

AU - Demello, Andrew

PY - 2009/9/24

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N2 - We report a method for removing unwanted contributions to fluorescence signals from dyes absorbed in polydimethylsiloxane (PDMS) using fluorescence lifetime imaging microscopy (FLIM). By analysing experimental fluorescence decays using a bi-exponential decay model, we are able to discriminate between emission originating from dye molecules in free solution and those absorbed within the PDMS substrate. Simple image processing allows the unwanted background signal to be removed and thus enables a more accurate assessment of temperature. The efficacy of the approach is demonstrated by measuring temperature changes within a droplet-based PCR device.

AB - We report a method for removing unwanted contributions to fluorescence signals from dyes absorbed in polydimethylsiloxane (PDMS) using fluorescence lifetime imaging microscopy (FLIM). By analysing experimental fluorescence decays using a bi-exponential decay model, we are able to discriminate between emission originating from dye molecules in free solution and those absorbed within the PDMS substrate. Simple image processing allows the unwanted background signal to be removed and thus enables a more accurate assessment of temperature. The efficacy of the approach is demonstrated by measuring temperature changes within a droplet-based PCR device.

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JO - Lab on a chip

JF - Lab on a chip

IS - 23

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Removal of background signals from fluorescence thermometry measurements in PDMS microchannels using fluorescence lifetime imaging

Abstract

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