Cellular Barcodes for Efficiently Profiling Single-Cell Secretory Responses by Microengraving

We present a method that uses fluorescent cellular barcodes to increase the number of unique samples that can be analyzed simultaneously by microengraving, a nanowell array-based technique for quantifying the secretory responses of thousands of single cells in parallel. Using n different fluorescent...

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Main Authors: Gierahn, Todd Michael (Contributor), Forcier, Talitha L. (Contributor), Brefo, Mavis S. (Contributor), Yamanaka, Yvonne J. (Author), Szeto, Gregory (Author), Arnold, Kelly B. (Author), Lauffenburger, Douglas A (Author), Irvine, Darrell J (Author), Love, John C (Author)
Other Authors: Massachusetts Institute of Technology. Department of Biological Engineering (Contributor), Massachusetts Institute of Technology. Department of Chemical Engineering (Contributor), Massachusetts Institute of Technology. Department of Materials Science and Engineering (Contributor), Ragon Institute of MGH, MIT and Harvard (Contributor), Koch Institute for Integrative Cancer Research at MIT (Contributor), Love, J. Christopher (Contributor), Yamanaka, Yvonne Joy (Contributor), Szeto, Gregory Lee (Contributor), Benedict, Kelly (Contributor), Lauffenburger, Douglas A. (Contributor), Irvine, Darrell J. (Contributor)
Format: Article
Language:English
Published: American Chemical Society, 2013-07-26T15:15:15Z.
Subjects:
Online Access:Get fulltext
LEADER 03546 am a22005053u 4500
001 79706
042 |a dc 
100 1 0 |a Gierahn, Todd Michael  |e author 
100 1 0 |a Massachusetts Institute of Technology. Department of Biological Engineering  |e contributor 
100 1 0 |a Massachusetts Institute of Technology. Department of Chemical Engineering  |e contributor 
100 1 0 |a Massachusetts Institute of Technology. Department of Materials Science and Engineering  |e contributor 
100 1 0 |a Ragon Institute of MGH, MIT and Harvard  |e contributor 
100 1 0 |a Koch Institute for Integrative Cancer Research at MIT  |e contributor 
100 1 0 |a Love, J. Christopher  |e contributor 
100 1 0 |a Yamanaka, Yvonne Joy  |e contributor 
100 1 0 |a Szeto, Gregory Lee  |e contributor 
100 1 0 |a Gierahn, Todd Michael  |e contributor 
100 1 0 |a Forcier, Talitha L.  |e contributor 
100 1 0 |a Benedict, Kelly  |e contributor 
100 1 0 |a Brefo, Mavis S.  |e contributor 
100 1 0 |a Lauffenburger, Douglas A.  |e contributor 
100 1 0 |a Irvine, Darrell J.  |e contributor 
700 1 0 |a Forcier, Talitha L.  |e author 
700 1 0 |a Brefo, Mavis S.  |e author 
700 1 0 |a Yamanaka, Yvonne J.  |e author 
700 1 0 |a Szeto, Gregory  |e author 
700 1 0 |a Arnold, Kelly B.  |e author 
700 1 0 |a Lauffenburger, Douglas A  |e author 
700 1 0 |a Irvine, Darrell J  |e author 
700 1 0 |a Love, John C  |e author 
245 0 0 |a Cellular Barcodes for Efficiently Profiling Single-Cell Secretory Responses by Microengraving 
260 |b American Chemical Society,   |c 2013-07-26T15:15:15Z. 
856 |z Get fulltext  |u http://hdl.handle.net/1721.1/79706 
520 |a We present a method that uses fluorescent cellular barcodes to increase the number of unique samples that can be analyzed simultaneously by microengraving, a nanowell array-based technique for quantifying the secretory responses of thousands of single cells in parallel. Using n different fluorescent dyes to generate 2n unique cellular barcodes, we achieved a 2n-fold reduction in the number of arrays and quantity of reagents required per sample. The utility of this approach was demonstrated in three applications of interest in clinical and experimental immunology. Using barcoded human peripheral blood mononuclear cells and T cells, we constructed dose-response curves, profiled the secretory behavior of cells treated with mechanistically distinct stimuli, and tracked the secretory behaviors of different lineages of CD4+ T helper cells. In addition to increasing the number of samples analyzed by generating secretory profiles of single cells from multiple populations in a time- and reagent-efficient manner, we expect that cellular barcoding in combination with microengraving will facilitate unique experimental opportunities for quantitatively analyzing interactions among heterogeneous cells isolated in small groups (2-5 cells). 
520 |a Howard Hughes Medical Institute (Investigator) 
520 |a National Institute of Allergy and Infectious Diseases (U.S.) (Grant 1R56AI104274) 
520 |a National Institute of Allergy and Infectious Diseases (U.S.) (Grant 1U19AI089992) 
520 |a National Institute of Allergy and Infectious Diseases (U.S.) (Grant 5U01AI068618) 
520 |a National Science Foundation (U.S.) (Fellowship) 
520 |a Massachusetts Institute of Technology (Collamore-Rogers Fellowship) 
520 |a Ragon Institute of MGH, MIT and Harvard 
520 |a W. M. Keck Foundation 
546 |a en_US 
655 7 |a Article 
773 |t Analytical Chemistry