Fluorescence lifetime biosensing with DNA microarrays and a CMOS-SPAD imager

Giraud, Gerard and Schulze, Holger and Li, Day-Uei and Bachmann, Till T. and Crain, Jason and Tyndall, David and Richardson, Justin and Walker, Richard and Stoppa, David and Charbon, Edoardo and Henderson, Robert and Arlt, Jochen (2010) Fluorescence lifetime biosensing with DNA microarrays and a CMOS-SPAD imager. Biomedical Optics Express. pp. 1302-1308. ISSN 2156-7085 (https://doi.org/10.1364/BOE.1.001302)

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Abstract

Fluorescence lifetime of dye molecules is a sensitive reporter on local microenvironment which is generally independent of fluorophores concentration and can be used as a means of discrimination between molecules with spectrally overlapping emission. It is therefore a potentially powerful multiplexed detection modality in biosensing but requires extremely low light level operation typical of biological analyte concentrations, long data acquisition periods and on-chip processing capability to realize these advantages. We report here fluorescence lifetime data obtained using a CMOS-SPAD imager in conjunction with DNA microarrays and TIRF excitation geometry. This enables acquisition of single photon arrival time histograms for a 320 pixel FLIM map within less than 26 seconds exposure time. From this, we resolve distinct lifetime signatures corresponding to dye-labelled HCV and quantum-dot-labelled HCMV nucleic acid targets at concentrations as low as 10 nM.

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