Issue 23, 2009
From the journal:

Lab on a Chip

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

Tom Robinson,ae   Yolanda Schaerli,b   Robert Wootton,b   Florian Hollfelder,b   Christopher Dunsby,c   Geoff Baldwin,d   Mark Neil,c   Paul French*c  and  Andrew deMello*e  

* Corresponding authors

a Chemical Biology Centre, Imperial College London, Exhibition Road, London, UK

b Department of Biochemistry, University of Cambridge, 80 Tennis Court Road, Cambridge, UK

c Photonics Group, Department of Physics, Exhibition Road, London, UK
E-mail: paul.french@imperial.ac.uk

d Division of Molecular Biosciences, Exhibition Road, London, UK

e Department of Chemistry, Exhibition Road, London, UK
E-mail: a.demello@imperial.ac.uk

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.

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

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Article information

DOI
https://doi.org/10.1039/B913293G
Article type
Technical Note
Submitted
06 Jul 2009
Accepted
25 Aug 2009
First published
24 Sep 2009

Lab Chip, 2009,9, 3437-3441

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

T. Robinson, Y. Schaerli, R. Wootton, F. Hollfelder, C. Dunsby, G. Baldwin, M. Neil, P. French and A. deMello, Lab Chip, 2009, 9, 3437 DOI: 10.1039/B913293G

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