Journal
NATURE PROTOCOLS
Volume 10, Issue 2, Pages 316-333Publisher
NATURE PUBLISHING GROUP
DOI: 10.1038/nprot.2015.020
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Funding
- US National Institutes of Health (NIH) [U19 AI057229, U54CA149145, N01-HV-00242, 1U19AI100627, 5R01AI07372405, R01CA184968, 1 R33 CA183654, NRSA F32 GM093508-01, 152175.5041015.0412, R01 A1073724]
- NIH-the Baylor Research Institute [41000411217]
- NIH-Northrop Grumman Corp. [7500108142]
- California Institute for Regenerative Medicine (CIRM) [DR1-01477, RB2-01592]
- US Department of Defense [OC110674]
- European Commission [Health.2010.1.2-1]
- US Food and Drug Administration [HHSF223201210194C]
- Bill and Melinda Gates Foundation [OPP 1017093]
- Alliance for Lupus Research, the Lymphoma Research Foundation
- Entertainment Industry Foundation (National Women's Cancer Research Alliance grant)
- National Science Foundation CAREER award [MCB-1149728]
- Packard Fellowship for Science and Engineering
- Rachford and Carlota A. Harris Endowed Professorship
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Mass-tag cell barcoding (MCB) labels individual cell samples with unique combinatorial barcodes, after which they are pooled for processing and measurement as a single multiplexed sample. The MCB method eliminates variability between samples in antibody staining and instrument sensitivity, reduces antibody consumption and shortens instrument measurement time. Here we present an optimized MCB protocol. The use of palladium-based labeling reagents expands the number of measurement channels available for mass cytometry and reduces interference with lanthanide-based antibody measurement. An error-detecting combinatorial barcoding scheme allows cell doublets to be identified and removed from the analysis. A debarcoding algorithm that is single cell-based rather than population-based improves the accuracy and efficiency of sample deconvolution. This debarcoding algorithm has been packaged into software that allows rapid and unbiased sample deconvolution. The MCB procedure takes 3-4 h, not including sample acquisition time of similar to 1 h per million cells.
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