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Title of Journal: Biotechnol Lett

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Abbravation: Biotechnology Letters

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Springer Netherlands

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DOI

10.1007/bfb0027417

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ISSN

1573-6776

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Singlecolour flow cytometric assay to determine N

Authors: Ajit Thakur Abeyat Zaman Jeff Hummel Kim Jones Gonzalo Hortelano
Publish Date: 2011/12/21
Volume: 34, Issue: 3, Pages: 447-453
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Abstract

A flow cytometrybased cytotoxicity FCC assay was developed using a single fluorophore calceinacetoxymethyl diacetylester calceinAM to measure NK cellmediated cytotoxicity Nonadherent human K562 and U937 target cells were individually labelled with calceinAM and coincubated with effector NK cells to measure calcein loss and therefore calculate target cell cytotoxicity This FCC assay also provided a measure of sample viability Notably cell viability measured by traditional calcein/7aminoactinomycin D 7AAD double labelling and Trypan Blue methods were comparable to the viability calculated using calceinloss FCC This FCC assay may also be used with various effector and target cell types and as a multiparameter tool to measure viability and immunophenotype cells for tissue engineering purposesWe would like to thank Dr Heather Sheardown Dr Karen Mossman Dr Ali Ashkar Dr Firoz Mian Dr Iran Rashedi Glenn McClung Ruchira Sengupta Faheem Dinath and Dave Morrison for providing us with materials and technical support and to the Natural Science and Engineering Research Council NSERC for their funding to AT and KJ

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