A novel flash detection algorithm for single molecule counting with TIRF microscopy

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dc.contributor.authorRadford, J.
dc.contributor.authorWang, L.
dc.contributor.authorLi, J.
dc.contributor.authorCoad, B.
dc.contributor.authorMcFarland, C.
dc.contributor.authorNordon, R.
dc.contributor.conference2013 IEEE 10th International Symposium on Biomedical Imaging (ISBI 2013) (7 Apr 2013 - 11 Apr 2013 : San Francisco, California)
dc.date.issued2013
dc.description.abstractA 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.
dc.description.statementofresponsibilityJoseph Radford, Liyuan Wang, JingJing Li, Bryan R. Coad, Clive D. McFarland, Robert E. Nordon
dc.identifier.citationProceedings / IEEE International Symposium on Biomedical Imaging: from nano to macro. IEEE International Symposium on Biomedical Imaging, 2013, pp.1066-1069
dc.identifier.doi10.1109/ISBI.2013.6556662
dc.identifier.isbn9781467364546
dc.identifier.issn1945-7928
dc.identifier.issn1945-8452
dc.identifier.orcidCoad, B. [0000-0003-3297-3977]
dc.identifier.urihttp://hdl.handle.net/2440/108641
dc.language.isoen
dc.publisherIEEE
dc.relation.granthttp://purl.org/au-research/grants/nhmrc/631931
dc.relation.ispartofseriesIEEE International Symposium on Biomedical Imaging
dc.rights© 2013 IEEE
dc.source.urihttps://doi.org/10.1109/isbi.2013.6556662
dc.subjectTIRF-M, single molecule imaging, protein adsorption, flash detection
dc.titleA novel flash detection algorithm for single molecule counting with TIRF microscopy
dc.typeConference paper
pubs.publication-statusPublished

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