Removal of background signals from fluorescence thermometry measurements in PDMS microchannels using fluorescence lifetime imaging.
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.
Robinson
Tom
T
Chemical Biology Centre, Imperial College London, Exhibition Road, London, SW7 2AZ, UK.
Schaerli
Yolanda
Y
Wootton
Robert
R
Hollfelder
Florian
F
Dunsby
Christopher
C
Baldwin
Geoff
G
Neil
Mark
M
French
Paul
P
deMello
Andrew
A
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
D004129:Dimethylpolysiloxanes; D005456:Fluorescent Dyes; D012235:Rhodamines; C029773:rhodamine B