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dc.contributor.authorRiddell, Andrewen
dc.contributor.authorGardner, Ruien
dc.contributor.authorPerez-Gonzalez, Alexisen
dc.contributor.authorLopes, Telmaen
dc.contributor.authorMartinez, Lolaen
dc.date.accessioned2015-05-21T12:33:43Z
dc.date.available2015-05-21T12:33:43Z
dc.date.issued2015-03-04en
dc.identifier.citationRiddell et al. Methods (2015) Vol. 82, pp. 64-73. pii: S1046-2023(15)00090-0. doi: 10.1016/j.ymeth.2015.02.017.en
dc.identifier.issn1046-2023
dc.identifier.urihttps://www.repository.cam.ac.uk/handle/1810/247902
dc.description.abstractSorting performance can be evaluated with regard to Purity, Yield and/or Recovery of the sorted fraction. Purity is a check on the quality of the sample and the sort decisions made by the instrument. Recovery and Yield definitions vary with some authors regarding both as how efficient the instrument is at sorting the target particles from the original sample, others distinguishing Recovery from Yield, where the former is used to describe the accuracy of the instrument’s sort count. Yield and Recovery are often neglected, mostly due to difficulties in their measurement. Purity of the sort product is often cited alone but is not sufficient to evaluate sorting performance. All of these three performance metrics require re-sampling of the sorted fraction. But, unlike Purity, calculating Yield and/or Recovery calls for the absolute counting of particles in the sorted fraction, which may not be feasible, particularly when dealing with rare populations and precious samples. In addition, the counting process itself involves large errors. Here we describe a new metric for evaluating instrument sort Recovery, defined as the number of particles sorted relative to the number of original particles to be sorted. This calculation requires only measuring the ratios of target and non-target populations in the original pre-sort sample and in the waste stream or center stream catch (CSC), avoiding re-sampling the sorted fraction and absolute counting. We called this new metric Rmax, since it corresponds to the maximum expected Recovery for a particular set of instrument parameters. Rmax is ideal to evaluate and troubleshoot the optimum drop-charge delay of the sorter, or any instrument related failures that will affect sort performance. It can be used as a daily quality control check but can be particularly useful to assess instrument performance before single-cell sorting experiments. Because we do not perturb the sort fraction we can calculate Rmax during the sort process, being especially valuable to check instrument performance during rare population sorts.
dc.description.sponsorshipAndrew Riddell is based at the Wellcome Trust-MRC Stem Cell Institute, Centre for Stem Cell Research University of Cambridge
dc.languageEnglishen
dc.language.isoenen
dc.publisherElsevier
dc.rightsAttribution 2.0 UK: England & Wales*
dc.rights.urihttp://creativecommons.org/licenses/by/2.0/uk/*
dc.subjectFlow cytometryen
dc.subjectRmaxen
dc.subjectRecoveryen
dc.subjectPurityen
dc.subjectYielden
dc.subjectDrop-charge delayen
dc.subjectSorten
dc.subjectInstrument performanceen
dc.subjectQuality controlen
dc.titleRmax: A systematic approach to evaluate instrument sort performance using center stream catchen
dc.typeArticle
dc.description.versionThis is the final published version. It first appeared at http://www.sciencedirect.com/science/article/pii/S1046202315000900en
prism.endingPage73
prism.publicationDate2015en
prism.publicationNameMethodsen
prism.startingPage64
prism.volume82en
dc.rioxxterms.funderWellcome Trust
dc.rioxxterms.funderMRC
dcterms.dateAccepted2015-02-25en
rioxxterms.versionofrecord10.1016/j.ymeth.2015.02.017en
rioxxterms.licenseref.urihttp://www.rioxx.net/licenses/all-rights-reserveden
rioxxterms.licenseref.startdate2015-03-04en
dc.identifier.eissn1095-9130
rioxxterms.typeJournal Article/Reviewen


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Attribution 2.0 UK: England & Wales
Except where otherwise noted, this item's licence is described as Attribution 2.0 UK: England & Wales