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A compact long-wavelength fluorescence detection system and its application to flow injection immunoassay
journal contributionposted on 08.08.2016 by Sijung Hu, Martin T. French, Derek A. Palmer, Mark Evans, Sue M. Zhou, G. Hussain Sarpara, James N. Miller
Any type of content formally published in an academic journal, usually following a peer-review process.
A compact and robust long-wavelength fluorescence detection system (LWFDS) has been constructed using a pulsed diode laser as the light source and a photodiode detector. The detection limits for naphthofluorescein (NF) and the cyanine dyes Cy5 and Cy5.5 at pH 8.8, determined using the LWFDS, were 1.0 × 10-10, 2.3 × 10-12 and 6.2 × 10-11M, respectively. These values compared favourably with data from a commercial research grade fluorescence spectrometer. The LWFDS instrument was used as the detector for a competitive heterogeneous flow injection immunoassay for α-interferon with Cy5 as the label. A calibration graph usable over the range 0.5-10 μg ml-1 α-interferon was obtained. The instrument demonstrated its potential for rapid ( <200s per sample) multi-analyte assays and for field monitoring.
S.J.H. acknowledges financial support from Loughborough University and from Kalibrant Limited. We also thank Glaxo-Wellcome for gifts of α-interferon and its antibodies.
- Mechanical, Electrical and Manufacturing Engineering