title: Removal of background signals from fluorescence thermometry measurements in PDMS microchannels using fluorescence lifetime imaging.
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.
authors.affiliation: Chemical Biology Centre, Imperial College London, Exhibition Road, London, SW7 2AZ, UK.
authors.forename: Tom
authors.initials: T
authors.lastname: Robinson
authors.forename: Yolanda
authors.initials: Y
authors.lastname: Schaerli
authors.forename: Robert
authors.initials: R
authors.lastname: Wootton
authors.forename: Florian
authors.initials: F
authors.lastname: Hollfelder
authors.forename: Christopher
authors.initials: C
authors.lastname: Dunsby
authors.forename: Geoff
authors.initials: G
authors.lastname: Baldwin
authors.forename: Mark
authors.initials: M
authors.lastname: Neil
authors.forename: Paul
authors.initials: P
authors.lastname: French
authors.forename: Andrew
authors.initials: A
authors.lastname: deMello
mesh_terms: D000327:Adsorption; D004129:Dimethylpolysiloxanes / Q000737:chemistry*; D004867:Equipment Design; D005453:Fluorescence; D005456:Fluorescent Dyes / Q000032:analysis*; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation* / Q000379:methods; D018613:Microscopy, Confocal / Q000379:methods; D012235:Rhodamines / Q000032:analysis*; D013696:Temperature
chemical_list: D004129:Dimethylpolysiloxanes; D005456:Fluorescent Dyes; D012235:Rhodamines; C029773:rhodamine B