8/13/2009 - ANNOUNCEMENT
Multiplexed labeling of samples with cell tracking dyes facilitates rapid and accurate internally controlled calcium flux measurement by flow cytometry.
Guo FuCorresponding Author Contact Information, a, E-mail The Corresponding Author and Nicholas R.J. GascoigneCorresponding Author Contact Information, a, E-mail The Corresponding Author
aDepartment of Immunology and Microbial Science, IMM1, The Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, California 92037, USA
Received 9 May 2009;
revised 21 July 2009;
accepted 21 July 2009.
Available online 30 July 2009.
Abstract
Calcium flux measurement is a crucial assay in lymphocyte activation. However, with the currently well established flow cytometric methods, it is a tedious procedure that is difficult to control to avoid variation between samples. This leads to unwanted sources of error that can make it problematic to interpret the results. Here we present an improved method that allows different cell populations to be tested in the same sample. Samples are pre-labeled with CFSE or Cy5 then mixed and stimulated to induce calcium flux. This facilitates more rapid and accurate measurement of calcium flux and also dramatically reduces the cost and effort required for this type of assay.
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