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An optical counting technique with vertical hydrodynamic focusing for biological cells

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dc.contributor.author Chiavaroli, Stefano
dc.contributor.author Newport, David
dc.contributor.author Woulfe, Bernie
dc.date.accessioned 2013-04-11T16:11:40Z
dc.date.available 2013-04-11T16:11:40Z
dc.date.issued 2010
dc.identifier.uri http://hdl.handle.net/10344/3032
dc.description peer-reviewed en_US
dc.description.abstract A barrier in scaling laboratory processes into automated microfluidic devices has been the transfer of lab based assays: where engineering meets biological protocol. One basic requirement is to reliably and accurately know the distribution and number of biological cells being dispensed. In this study, a novel optical counting technique to efficiently quantify the number of cells flowing into a microtube is presented. REH, B-lymphoid precursor leukaemia, are stained with a fluorescent dye and frames of moving cells are recorded using a CCD camera. The basic principle is to calculate the total fluorescence intensity of the image and to divide it by the average intensity of a single cell. This method allows counting the number of cells with an uncertainty +/- 5%, which compares favourably to the standard biological methodology, based on the manual Trypan Blue assay, which is destructive to the cells and presents an uncertainty in the order of 20%. The use of a microdevice for vertical hydrodynamic focusing, which can reduce the background noise of out of focus cells by concentrating the cells in a thin layer, has further improved the technique. CFD simulation and Confocal Laser Scanning Microscopy images have shown an 82% reduction in the vertical displacement of the cells. For the flow rates imposed during this study, a throughput of 100-200 cells/sec is achieved. en_US
dc.language.iso eng en_US
dc.publisher American Institute of Physics en_US
dc.relation.ispartofseries Biomicrofluidics;4(2), 024110
dc.relation.uri http://dx.doi.org/10.1063/1.3380598
dc.rights Copyright 2010 American Institute of Physics. This article may be downloaded for personal use only. Any other use requires prior permission of the author and the American Institute of Physics. The following article appeared in Biomicrofluidics, 2010, 4(2), 024110 and may be found at http://dx.doi.org/10.1063/1.3380598 en_US
dc.subject optical counting technique en_US
dc.subject engineering en_US
dc.subject biological cells en_US
dc.title An optical counting technique with vertical hydrodynamic focusing for biological cells en_US
dc.type info:eu-repo/semantics/article en_US
dc.type.supercollection all_ul_research en_US
dc.type.supercollection ul_published_reviewed en_US
dc.contributor.sponsor European Commission under the Marie Curie Early Stage Training Fellowship en_US
dc.contributor.sponsor Mid-Western Cancer Foundation, Limerick, Ireland en_US
dc.relation.projectid MEST-CT-2004-505101 en_US
dc.rights.accessrights info:eu-repo/semantics/openAccess en_US
dc.internal.rssid 1131913


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