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Type: Conference paper
Title: A novel flash detection algorithm for single molecule counting with TIRF microscopy
Author: Radford, J.
Wang, L.
Li, J.
Coad, B.
McFarland, C.
Nordon, R.
Citation: Proceedings / IEEE International Symposium on Biomedical Imaging: from nano to macro. IEEE International Symposium on Biomedical Imaging, 2013, pp.1066-1069
Publisher: IEEE
Issue Date: 2013
Series/Report no.: IEEE International Symposium on Biomedical Imaging
ISBN: 9781467364546
ISSN: 1945-7928
Conference Name: 2013 IEEE 10th International Symposium on Biomedical Imaging (ISBI 2013) (7 Apr 2013 - 11 Apr 2013 : San Francisco, California)
Statement of
Joseph Radford, Liyuan Wang, JingJing Li, Bryan R. Coad, Clive D. McFarland, Robert E. Nordon
Abstract: A novel algorithm, Adjacent Pixel Temporal Intensity Correlation (APTIC), was developed to detect single fluorescent molecules by their stochastic emission patterns; photoblinking and photobleaching. The algorithm was evaluated using simulated image data and Total Internal Reflection Fluorescence Microscopy (TIRF-M) to count the number fluorescently labelled protein molecules adsorbed onto glass substrates modified by Radio Frequency Glow Discharge (RFGD) deposition. By selecting an appropriate correlation threshold, the algorithm was capable of detecting synthetic flashes with a signal-to-noise ratio (SNR) as low as 2.0 with 90% sensitivity. The methodology holds great promise for mapping the amount and distribution of biomolecules on surfaces.
Keywords: TIRF-M, single molecule imaging, protein adsorption, flash detection
Rights: © 2013 IEEE
DOI: 10.1109/ISBI.2013.6556662
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