A clinical microchip for evaluation of single immune cells reveals high functional heterogeneity in phenotypically similar T cells
Open accessAbstract
Cellular immunity has an inherent high level of functional heterogeneity. Capturing the full spectrum of these functions requires analysis of large numbers of effector molecules from single cells. We report a microfluidic platform designed for highly multiplexed (more than ten proteins), reliable, sample-efficient (∼1 × 104 cells) and quantitative measurements of secreted proteins from single cells. We validated the platform by assessment of multiple inflammatory cytokines from lipopolysaccharide (LPS)-stimulated human macrophages and comparison to standard immunotechnologies. We applied the platform toward the ex vivo quantification of T cell polyfunctional diversity via the simultaneous measurement of a dozen effector molecules secreted from tumor antigen–specific cytotoxic T lymphocytes (CTLs) that were actively responding to tumor and compared against a cohort of healthy donor controls. We observed profound, yet focused, functional heterogeneity in active tumor antigen–specific CTLs, with the major functional phenotypes quantitatively identified. The platform represents a new and informative tool for immune monitoring and clinical assessment.
The claims and statistics of this paper haven’t been checked yet.
Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.
Integrity signals
Registry
Cites
Statistics
Claims the paper makes
The claims and statistics of this paper haven’t been checked yet.
Run the check — free, no account needed. It reads the open-access text and checks the sources it cites.
Self-check
Percentages, sums and reported differences re-computed from the paper’s own numbers.
The self-check needs the paper’s full text.
Cites
Coussens, L. M., & Werb, Z. (2002) · Nature · doi:10.1038/nature01322 · Cited by 15,111 · see citing works
Cluster analysis and display of genome-wide expression patterns
Eisen, M. B., Spellman, P. T., Brown, P. O., & Botstein, D. (1998) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.95.25.14863 · Cited by 12,617 · see citing works
Mantovani, A., Allavena, P., Sica, A., & Balkwill, F. (2008) · Nature · doi:10.1038/nature07205 · Cited by 11,399 · see citing works
Origin and physiological roles of inflammation
Medzhitov, R. (2008) · Nature · doi:10.1038/nature07201 · Cited by 6,618 · see citing works
Monocyte and macrophage heterogeneity
Gordon, S., & Taylor, P. R. (2005) · Nature Reviews Immunology · doi:10.1038/nri1733 · Cited by 5,190 · see citing works
Toll-like receptors in the induction of the innate immune response
Aderem, A. A., & Ulevitch, R. J. (2000) · Nature · doi:10.1038/35021228 · Cited by 3,047 · see citing works
Cancer regression in patients after transfer of genetically engineered lymphocytes
Morgan, R. A., Dudley, M. E., Wunderlich, J. R., Hughes, M. S., Yang, J. C., Sherry, R. M., Royal, R. E., Topalian, S. L., Kammula, U. S., Restifo, N. P., et al. (2006) · Science · doi:10.1126/science.1129003 · Cited by 2,603 · see citing works
Microfluidic Large-Scale Integration
Thorsen, T., Maerkl, S. J., & Quake, S. R. (2002) · Science · doi:10.1126/science.1076996 · Cited by 2,223 · see citing works
T-cell quality in memory and protection: implications for vaccine design
Seder, R. A., Darrah, P. A., & Roederer, M. (2008) · Nature reviews. Immunology · doi:10.1038/nri2274 · Cited by 1,541 · see citing works
Multifunctional TH1 cells define a correlate of vaccine-mediated protection against Leishmania major
Darrah, P. A., Patel, D. T., Luca, P. M. D., Lindsay, R. W. B., Davey, D. F., Flynn, B. J., Hoff, S. T., Andersen, P. L., Reed, S. G., Morris, S. L., et al. (2007) · Nature Medicine · doi:10.1038/nm1592 · Cited by 1,364 · see citing works
Characterization of circulating T cells specific for tumor-associated antigens in melanoma patients
Lee, P. P., Yee, C., Savage, P. A., Fong, L. H., Brockstedt, D. G., Weber, J. S., Johnson, D. L., Swetter, S. M., Thompson, J. A., Greenberg, P. D., et al. (1999) · Nature Medicine · doi:10.1038/9525 · Cited by 1,117 · see citing works
Phenotype and function of human T lymphocyte subsets: Consensus and issues
Appay, V., van Lier, R. A. W., Sallusto, F., & Roederer, M. (2008) · Cytometry Part A · doi:10.1002/cyto.a.20643 · Cited by 729 · cited 1× · see citing works
“Viral infection leads to activated T cells with a large variety of effector functions, as reflected by different cytokine profile”
Kaech, S. M., & Wherry, E. J. (2007) · Immunity · doi:10.1016/j.immuni.2007.08.007 · Cited by 619 · see citing works
Fan, R., Vermesh, O., Srivastava, A., Yen, B. K. H., Qin, L., Ahmad, H. M. N., Kwong, G. A., Liu, C.-C., Gould, J., Hood, L., et al. (2008) · Nature Biotechnology · doi:10.1038/nbt.1507 · Cited by 539 · see citing works
Self-citationPrecopio, M., Betts, M. R., Parrino, J., Price, D. A., Emma Gostick, D., Ambrozak, D. R., Asher, T. E., Douek, D. C., Harari, A., Pantaleo, G., et al. (2007) · The Journal of Experimental Medicine · doi:10.1084/jem.20062363 · Cited by 484 · see citing works
Pantaleo, G., & Koup, R. A. (2004) · Nature Medicine · doi:10.1038/nm0804-806 · Cited by 445 · see citing works
A live-cell assay to detect antigen-specific CD4+ T cells with diverse cytokine profiles
Chattopadhyay, P. K., Yu, J., & Roederer, M. (2005) · Nature Medicine · doi:10.1038/nm1293 · Cited by 321 · see citing works
Human CD8+ T-cell differentiation in response to viruses
van Lier, R. A. W., ten Berge, I. J. M., & Gamadia, L. E. (2003) · Nature Reviews Immunology · doi:10.1038/nri1254 · Cited by 297 · see citing works
Bachmann, M. F., Wolint, P., Schwarz, K., Jäger, P., & Oxenius, A. (2005) · The Journal of Immunology · doi:10.4049/jimmunol.175.7.4686 · Cited by 274 · see citing works
Bailey, R. C., Kwong, G. A., Radu, C. G., Witte, O. N., & Heath, J. R. (2007) · Journal of the American Chemical Society · doi:10.1021/ja065930i · Cited by 272 · see citing works
Self-citationFan, J., & Malik, A. B. (2003) · Nature Medicine · doi:10.1038/nm832 · Cited by 265 · see citing works
Kotecha, N., Flores, N. J., Irish, J. M., Simonds, E. F., Sakai, D. S., Archambeault, S., Díaz-Flores, E., Coram, M., Shannon, K., Nolan, G. P., et al. (2008) · Cancer Cell · doi:10.1016/j.ccr.2008.08.014 · Cited by 233 · see citing works
Dendritic Cell Vaccination Combined with CTLA4 Blockade in Patients with Metastatic Melanoma
Ribas, A., Comı́n-Anduix, B., Chmielowski, B., Jalil, J., de la Rocha, P., McCannel, T. A., Ochoa, M. T., Seja, E., Villanueva, A., Oseguera, D. K., et al. (2009) · Clinical Cancer Research · doi:10.1158/1078-0432.ccr-09-1254 · Cited by 217 · see citing works
Self-citationVaccination in Humans Generates Broad T Cell Cytokine Responses
Rosa, S. C. D., Lü, F. X.-S., Yu, J., Perfetto, S. P., Falloon, J., Moser, S., Evans, T. G., Koup, R. A., Miller, C. J., & Roederer, M. (2004) · The Journal of Immunology · doi:10.4049/jimmunol.173.9.5372 · Cited by 204 · see citing works
Systems biology of innate immunity
Zak, D. E., & Aderem, A. A. (2008) · Immunological Reviews · doi:10.1111/j.1600-065x.2008.00721.x · Cited by 153 · see citing works
Inflammation and Cancer: A Double-Edged Sword
Hagemann, T., Balkwill, F. R., & Lawrence, T. (2007) · Cancer Cell · doi:10.1016/j.ccr.2007.10.005 · Cited by 152 · see citing works
Makedonas, G., & Betts, M. R. (2006) · Springer Seminars in Immunopathology · doi:10.1007/s00281-006-0025-4 · Cited by 126 · see citing works
Attig, S., Hennenlotter, J., Pawelec, G., Klein, G., Koch, S. D., Pircher, H., Feyerabend, S., Wernet, D., Stenzl, A., Rammensee, H., et al. (2009) · Cancer Research · doi:10.1158/0008-5472.can-09-0852 · Cited by 91 · see citing works
Wang, J., Ahmad, H. M. N., Ma, C., Shi, Q., Vermesh, O., Vermesh, U., & Heath, J. R. (2010) · Lab on a Chip · doi:10.1039/c0lc00132e · Cited by 90 · see citing works
Self-citationModular Nucleic Acid Assembled p/MHC Microarrays for Multiplexed Sorting of Antigen-Specific T Cells
Kwong, G. A., Radu, C. G., Hwang, K., Shu, C. J., Ma, C., Koya, R. C., Comin-Anduix, B., Hadrup, S. R., Bailey, R. C., Witte, O. N., et al. (2009) · Journal of the American Chemical Society · doi:10.1021/ja9006707 · Cited by 88 · see citing works
Self-citationSong, K., Rabin, R. L., Hill, B. J., Rosa, S. C. D., Perfetto, S. P., Zhang, H. H., Foley, J. F., Reiner, J. S., Liu, J., Mattapallil, J. J., et al. (2005) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.0409720102 · Cited by 87 · see citing works
Comı́n-Anduix, B., Lee, Y., Jalil, J., Algazi, A. P., de la Rocha, P., Camacho, L. H., Bozon, V. A., Bulanhagui, C. A., Seja, E., Villanueva, A., et al. (2008) · Journal of Translational Medicine · doi:10.1186/1479-5876-6-22 · Cited by 82 · see citing works
Self-citationHeterogeneity of TLR-induced responses in dendritic cells: from innate to adaptive immunity
Re, F., & Strominger, J. L. (2004) · Immunobiology · doi:10.1016/j.imbio.2004.03.005 · Cited by 73 · see citing works
Protein Signaling Networks from Single Cell Fluctuations and Information Theory Profiling
Shin, Y. S., Remacle, F., Fan, R., Hwang, K., Wei, W., Ahmad, H. M. N., Levine, R. D., & Heath, J. R. (2011) · Biophysical Journal · doi:10.1016/j.bpj.2011.04.025 · Cited by 56 · see citing works
Self-citationShin, Y. S., Ahmad, H. M. N., Shi, Q., Kim, H., Pascal, T. A., Fan, R., Goddard, W. A., & Heath, J. R. (2010) · ChemPhysChem · doi:10.1002/cphc.201000528 · Cited by 52 · see citing works
Self-citationComı́n-Anduix, B., Gualberto, A., Glaspy, J. A., Seja, E., Ontiveros, M., Reardon, D., Renteria, R., Englahner, B., Economou, J. S., Gómez-Navarro, J., et al. (2006) · Clinical Cancer Research · doi:10.1158/1078-0432.ccr-05-0136 · Cited by 41 · see citing works
Self-citationT cell heterogeneity: firmly fixed, predominantly plastic or merely malleable?
O’Shea, J. J., Hunter, C. A., & Germain, R. N. (2008) · Nature Immunology · doi:10.1038/ni0508-450 · Cited by 32 · see citing works
Flagged first, then by citations
1 of the 38 references couldn’t be matched to a known paper.
Cited by
Sorted by citations
Primary, Adaptive, and Acquired Resistance to Cancer Immunotherapy
Sharma, P., Hu-Lieskovan, S., Wargo, J. A., & Ribas, A. (2017) · Cell · doi:10.1016/j.cell.2017.01.017 · Cited by 5,588 · see citing works
Self-citationFunctional and Biomimetic Materials for Engineering of the Three-Dimensional Cell Microenvironment
Huang, G., Li, F., Zhao, X., Ma, Y., Li, Y., Lin, M., Jin, G., Lu, T. J., Genin, G. M., & Xu, F. (2017) · Chemical Reviews · doi:10.1021/acs.chemrev.7b00094 · Cited by 825 · see citing works
Newell, E. W., Sigal, N., Bendall, S. C., Nolan, G. P., & Davis, M. M. (2012) · Immunity · doi:10.1016/j.immuni.2012.01.002 · Cited by 555 · see citing works
Single-cell analysis tools for drug discovery and development
Heath, J. R., Ribas, A., & Mischel, P. S. (2016) · Nature Reviews Drug Discovery · doi:10.1038/nrd.2015.16 · Cited by 473 · see citing works
Self-citationBiosensor technology: recent advances in threat agent detection and medicine
Kirsch, J. S., Siltanen, C. A., Zhou, Q., Revzin, A., & Simonian, A. L. (2013) · Chemical Society Reviews · doi:10.1039/c3cs60141b · Cited by 463 · see citing works
Single-cell technologies for monitoring immune systems
Chattopadhyay, P. K., Gierahn, T. M., Roederer, M., & Love, J. C. (2014) · Nature Immunology · doi:10.1038/ni.2796 · Cited by 368 · see citing works
Microfluidics: reframing biological enquiry
Duncombe, T. A., Tentori, A. M., & Herr, A. E. (2015) · Nature Reviews Molecular Cell Biology · doi:10.1038/nrm4041 · Cited by 347 · see citing works
Zhao, J. L., Ma, C., O’Connell, R. M., Mehta, A., DiLoreto, R., Heath, J. R., & Baltimore, D. (2014) · Cell stem cell · doi:10.1016/j.stem.2014.01.007 · Cited by 333 · see citing works
Self-citationPolyfunctional responses by human T cells result from sequential release of cytokines
Han, Q., Bagheri, N., Bradshaw, E. M., Hafler, D. A., Lauffenburger, D. A., & Love, J. C. (2011) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.1117194109 · Cited by 329 · see citing works
Scaling by shrinking: empowering single-cell 'omics' with microfluidic devices
Prakadan, S. M., Shalek, A. K., & Weitz, D. A. (2017) · Nature Reviews Genetics · doi:10.1038/nrg.2017.15 · Cited by 327 · see citing works
Multiplex Serum Cytokine Immunoassay Using Nanoplasmonic Biosensor Microarrays
Chen, P., Chung, M. T., McHugh, W. M., Nidetz, R., Li, Y., Fu, J., Cornell, T. T., Shanley, T. P., & Kurabayashi, K. (2015) · ACS Nano · doi:10.1021/acsnano.5b00396 · Cited by 322 · see citing works
Single-cell proteomic chip for profiling intracellular signaling pathways in single tumor cells
Shi, Q., Qin, L., Wei, W., Geng, F., Fan, R., Shin, Y. S., Guo, D., Hood, L., Mischel, P. S., & Heath, J. R. (2011) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.1110865109 · Cited by 318 · see citing works
Self-citationKleppe, M., Kwak, M., Koppikar, P., Riester, M., Keller, M., Bastian, L., Hricik, T., Bhagwat, N., McKenney, A. S., Papalexi, E., et al. (2015) · Cancer Discovery · doi:10.1158/2159-8290.cd-14-0736 · Cited by 317 · see citing works
Self-citationMicro Total Analysis Systems for Cell Biology and Biochemical Assays
Kovarik, M. L., Gach, P. C., Ornoff, D. M., Wang, Y., Balowski, J., Farrag, L., & Allbritton, N. L. (2011) · Analytical Chemistry · doi:10.1021/ac202611x · Cited by 268 · see citing works
Lu, Y., Xue, Q., Eisele, M. R., Sulistijo, E., Brower, K., Han, L., Amir, E., Pe'er, D., Miller-Jensen, K., & Fan, R. (2015) · Proceedings of the National Academy of Sciences of the United States of America · doi:10.1073/pnas.1416756112 · Cited by 246 · see citing works
Self-citationSystems Biology and P4 Medicine: Past, Present, and Future
Hood, L. (2013) · Rambam Maimonides Medical Journal · doi:10.5041/rmmj.10112 · Cited by 267 · see citing works
Chodon, T., Comı́n-Anduix, B., Chmielowski, B., Koya, R. C., Wu, Z., Auerbach, M., Ng, C., Avramis, E. V., Seja, E., Villanueva, A., et al. (2014) · Clinical Cancer Research · doi:10.1158/1078-0432.ccr-13-3017 · Cited by 259 · see citing works
Self-citationChemical Analysis of Single Cells
Lin, Y., Trouillon, R., Рустамовна, С. Г., & Ewing, A. G. (2011) · Analytical Chemistry · doi:10.1021/ac2009838 · Cited by 251 · see citing works
Harnessing Nanomaterials for Cancer Sonodynamic Immunotherapy
Liang, S., Yao, J., Liu, D., Rao, L., Chen, X. S., & Wang, Z. (2023) · Advanced Materials · doi:10.1002/adma.202211130 · Cited by 245 · see citing works
Single-cell analysis targeting the proteome
Labib, M., & Kelley, S. O. (2020) · Nature Reviews Chemistry · doi:10.1038/s41570-020-0162-7 · Cited by 233 · see citing works
Microfluidic modelling of the tumor microenvironment for anti-cancer drug development
Shang, M., Soon, R. H., Lim, C. T., Khoo, B. L., & Han, J. (2019) · Lab on a Chip · doi:10.1039/c8lc00970h · Cited by 227 · see citing works
Microfluidic single cell analysis: from promise to practice
Lecault, V., White, A. K., Singhal, A., & Hansen, C. L. (2012) · Current Opinion in Chemical Biology · doi:10.1016/j.cbpa.2012.03.022 · Cited by 228 · see citing works
Microfluidics: A New Tool for Modeling Cancer–Immune Interactions
Boussommier-Calleja, A., Li, R., Chen, M. B., Wong, S. C., & Kamm, R. D. (2016) · Trends in Cancer · doi:10.1016/j.trecan.2015.12.003 · Cited by 224 · see citing works
“cytokine profiling, transcriptomics and proteomics of immune cells[76] [77]) to diagnose and tailor drug treatment[65], as reviewed elsewhere[45].”
BackgroundKleppe, M., Koche, R., Zou, L., van Galen, P., Hill, C. E., Dong, L., De Groote, S., Papalexi, E., Hanasoge Somasundara, A. V., Cordner, K., et al. (2018) · Cancer Cell · doi:10.1016/j.ccell.2017.11.009 · Cited by 279 · see citing works
Single cells make big data: New challenges and opportunities in transcriptomics
Angerer, P., Simon, L. M., Tritschler, S., Wolf, F., Fischer, D. S., & Theis, F. J. (2017) · Current Opinion in Systems Biology · doi:10.1016/j.coisb.2017.07.004 · Cited by 213 · see citing works
Kovarik, M. L., Ornoff, D. M., Melvin, A. T., Dobes, N. C., Wang, Y., Dickinson, A. J., Gach, P. C., Shah, P. K., & Allbritton, N. L. (2012) · Analytical Chemistry · doi:10.1021/ac3031543 · Cited by 219 · see citing works
Heterogeneity in immune responses: from populations to single cells
Satija, R., & Shalek, A. K. (2014) · Trends in Immunology · doi:10.1016/j.it.2014.03.004 · Cited by 210 · see citing works
Microfluidics for Manipulating Cells
Mu, X., Zheng, W., Sun, J., Zhang, W., & Jiang, X. (2012) · Small · doi:10.1002/smll.201200996 · Cited by 214 · see citing works
Microfluidic Single-Cell Manipulation and Analysis: Methods and Applications
Luo, T., Fan, L., Zhu, R., & Sun, D. B. (2019) · Micromachines · doi:10.3390/mi10020104 · Cited by 201 · see citing works
Hydrogel microparticles for biosensing
Goff, G. C. L., Srinivas, R. L., Hill, W. A., & Doyle, P. S. (2015) · European Polymer Journal · doi:10.1016/j.eurpolymj.2015.02.022 · Cited by 201 · see citing works
Oyler‐Yaniv, A., Oyler, J. E., Whitlock, B. M., Liu, Z., Germain, R. N., Huse, M., Altan-Bonnet, G. Y., & Krichevsky, O. (2017) · Immunity · doi:10.1016/j.immuni.2017.03.011 · Cited by 198 · see citing works
Digital Quantification of Proteins and mRNA in Single Mammalian Cells
Albayrak, C., Jordi, C. A., Zechner, C., Lin, J., Bichsel, C. A., Khammash, M., & Tay, S. (2016) · Molecular Cell · doi:10.1016/j.molcel.2016.02.030 · Cited by 176 · see citing works
Guo, H., Zhu, P., Guo, F., Li, X., Wu, X., Fan, X., Wen, L., & Tang, F. (2015) · Nature Protocols · doi:10.1038/nprot.2015.039 · Cited by 188 · see citing works
Multiomic analysis of cytokines in immuno-oncology
Jurišić, V. (2020) · Expert Review of Proteomics · doi:10.1080/14789450.2020.1845654 · Cited by 185 · see citing works
Chip in a lab: microfluidics for next generation life science research
Streets, A. M., & Huang, Y. (2013) · Biomicrofluidics · doi:10.1063/1.4789751 · Cited by 180 · see citing works
Self-Folding Single Cell Grippers
Malachowski, K., Jamal, M., Jin, Q., Polat, B., Morris, C. J., & Gracias, D. H. (2014) · Nano Letters · doi:10.1021/nl500136a · Cited by 177 · see citing works
Vistain, L. F., & Tay, S. (2021) · Trends in Biochemical Sciences · doi:10.1016/j.tibs.2021.01.013 · Cited by 177 · see citing works
High-Throughput Secretomic Analysis of Single Cells to Assess Functional Cellular Heterogeneity
Lu, Y., Chen, J. J. F., Mu, L., Xue, Q., Wu, Y., Wu, P.-H., Li, J., Vortmeyer, A. O., Miller‐Jensen, K., Wirtz, D., et al. (2013) · Analytical Chemistry · doi:10.1021/ac400082e · Cited by 173 · see citing works
Self-citationChemical Analysis of Single Cells
Trouillon, R., Passarelli, M. K., Wang, J., Kurczy, M. E., & Ewing, A. G. (2012) · Analytical Chemistry · doi:10.1021/ac303290s · Cited by 170 · see citing works
Wu, M., & Singh, A. K. (2011) · Current Opinion in Biotechnology · doi:10.1016/j.copbio.2011.11.023 · Cited by 167 · see citing works
Recent advances in single cell manipulation and biochemical analysis on microfluidics
Gao, D., Jin, F., Zhou, M., & Jiang, Y. (2018) · The Analyst · doi:10.1039/c8an01186a · Cited by 162 · see citing works
Functional TCR T cell screening using single-cell droplet microfluidics
Ségaliny, A. I., Li, G., Kong, L., Ren, C., Chen, X., Wang, J. K., Baltimore, D., Wu, G., & Zhao, W. (2018) · Lab on a Chip · doi:10.1039/c8lc00818c · Cited by 162 · see citing works
Recent advances in the use of microfluidic technologies for single cell analysis
Murphy, T. W., Zhang, Q., Naler, L. B., Ma, S., & Lu, C. (2017) · The Analyst · doi:10.1039/c7an01346a · Cited by 156 · see citing works
Nanotechnology: emerging tools for biology and medicinePreprint
Wong, I. Y., Bhatia, S. N., & Toner, M. (2013) · Genes & Development · doi:10.1101/gad.226837.113 · Cited by 150 · see citing works
Materials for Microfluidic Immunoassays: A Review
Mou, L., & Jiang, X. (2017) · Advanced Healthcare Materials · doi:10.1002/adhm.201601403 · Cited by 147 · see citing works
Ma, C., Cheung, A. F., Chodon, T., Koya, R. C., Wu, Z., Ng, C., Avramis, E. V., Cochran, A. J., Witte, O. N., Baltimore, D., et al. (2013) · Cancer Discovery · doi:10.1158/2159-8290.cd-12-0383 · Cited by 144 · see citing works
Self-citationXue, Q., Lu, Y., Eisele, M. R., Sulistijo, E. S., Khan, N., Fan, R., & Miller-Jensen, K. (2015) · Science Signaling · doi:10.1126/scisignal.aaa2155 · Cited by 143 · see citing works
Self-citationLiu, Y., Kwa, T., & Revzin, A. (2012) · Biomaterials · doi:10.1016/j.biomaterials.2012.06.089 · Cited by 141 · see citing works
Multiplexed profiling of single-cell extracellular vesicles secretion
Ji, Y., Qi, D., Li, L., Su, H., Li, X., Luo, Y., Sun, B., Zhang, F., Lin, B., Liu, T., et al. (2019) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.1814348116 · Cited by 138 · see citing works
Future of Digital Assays to Resolve Clinical Heterogeneity of Single Extracellular Vesicles
Morales, R. T., & Ko, J. (2022) · ACS Nano · doi:10.1021/acsnano.2c04337 · Cited by 135 · see citing works
Zhu, H., Zou, G., Wang, N., Zhuang, M., Xiong, W., & Huang, G. (2017) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.1615557114 · Cited by 133 · see citing works
Magnetophoretic circuits for digital control of single particles and cells
Lim, B., Reddy, V., Hu, X., Kim, K., Jadhav, M., Abedini‐Nassab, R., Noh, Y., Lim, Y. T., Yellen, B. B., & Kim, C. (2014) · Nature Communications · doi:10.1038/ncomms4846 · Cited by 131 · see citing works
Making a big thing of a small cell – recent advances in single cell analysis
Galler, K., Bräutigam, K., Große, C., Popp, J., & Neugebauer, U. (2014) · The Analyst · doi:10.1039/c3an01939j · Cited by 121 · see citing works
Digital microfluidic immunocytochemistry in single cells
Ng, A. H. C., Chamberlain, M. D., Situ, H., Lee, V., & Wheeler, A. R. (2015) · Nature Communications · doi:10.1038/ncomms8513 · Cited by 123 · see citing works
Real-time single-cell imaging of protein secretion
Shirasaki, Y., Yamagishi, M., Suzuki, N., Izawa, K., Nakahara, A., Mizuno, J., Shoji, S., Heike, T., Harada, Y., Nishikomori, R., et al. (2014) · Scientific Reports · doi:10.1038/srep04736 · Cited by 119 · see citing works
Microfluidic Single‐Cell Omics Analysis
Xu, X., Wang, J., Wu, L., Guo, J., Song, Y., Tian, T., Wang, W., Zhu, Z., & Yang, C. J. (2019) · Small · doi:10.1002/smll.201903905 · Cited by 120 · see citing works
Integrated Proteome Analysis Device for Fast Single-Cell Protein Profiling
Shao, X., Wang, X., Guan, S. U., Lin, H., Yan, G., Gao, M., Deng, C., & Zhang, X. (2018) · Analytical Chemistry · doi:10.1021/acs.analchem.8b03692 · Cited by 115 · see citing works
Remotely Activated Protein-Producing Nanoparticles
Schroeder, A., Goldberg, M. S., Kastrup, C. J., Wang, Y., Jiang, S., Joseph, B. J., Levins, C. G., Kannan, S. T., Langer, R., & Anderson, D. G. (2012) · Nano Letters · doi:10.1021/nl2036047 · Cited by 114 · see citing works
Recent Developments in Microfluidics for Cell Studies
Xiong, B., Ren, K., Shu, Y., Chen, Y., Shen, B., & Wu, H. (2014) · Advanced Materials · doi:10.1002/adma.201305348 · Cited by 112 · see citing works
Deng, Y., Zhang, Y., Sun, S., Wang, Z., Wang, M., Yu, B., Czajkowsky, D. M., Liu, B., Li, Y., Wei, W., et al. (2014) · Scientific Reports · doi:10.1038/srep07499 · Cited by 109 · see citing works
Self-citationBlom, K. G., Braun, M., Ivarsson, M. A., Gonzalez, V. D., Falconer, K., Moll, M., Ljunggren, H., Michaëlsson, J., & Sandberg, J. K. (2013) · The Journal of Immunology · doi:10.4049/jimmunol.1202234 · Cited by 108 · see citing works
Advances in Proteomic Techniques for Cytokine Analysis: Focus on Melanoma Research
Skalníková, H. K., Čížková, J., Červenka, J., & Vodička, P. (2017) · International Journal of Molecular Sciences · doi:10.3390/ijms18122697 · Cited by 105 · see citing works
Immune cell profiling in cancer: molecular approaches to cell-specific identification
Lyons, Y. A., Wu, S. Y., Overwijk, W. W., Baggerly, K., & Sood, A. K. (2017) · npj Precision Oncology · doi:10.1038/s41698-017-0031-0 · Cited by 104 · see citing works
Analytical techniques for multiplex analysis of protein biomarkers
van Gool, A. V., Corrales, F., Čolović, M., Krstić, D., Oliver-Martos, B., Martínez-Cáceres, E., Jakaša, I., Gajski, G., Brun, V., Kyriacou, K., et al. (2020) · Espert Review of Proteomics · doi:10.1080/14789450.2020.1763174 · Cited by 94 · see citing works
Son, K. J., Rahimian, A., Shin, D., Siltanen, C. A., Patel, T. C., & Revzin, A. (2015) · The Analyst · doi:10.1039/c5an01648g · Cited by 102 · see citing works
IL-33 fine tunes mast cell degranulation and chemokine production at the single-cell level
Joulia, R., L'Faqihi, F.-E., Valitutti, S., & Espinosa, É. (2016) · Journal of Allergy and Clinical Immunology · doi:10.1016/j.jaci.2016.09.049 · Cited by 101 · see citing works
Lü, Y., Ko, M. E., Cheng, H., Zhu, R., Xue, M., Wang, J. K., Lee, J. W., Frankiw, L., Xu, A. M., Wong, S. K., et al. (2020) · Nature Communications · doi:10.1038/s41467-020-15956-9 · Cited by 100 · see citing works
Self-citationRecent developments in microfluidic large scale integration
Araci, I. E., & Brisk, P. (2013) · Current Opinion in Biotechnology · doi:10.1016/j.copbio.2013.08.014 · Cited by 98 · see citing works
Microfluidic single-cell analysis for systems immunology
Junkin, M., & Tay, S. (2014) · Lab on a Chip · doi:10.1039/c3lc51182k · Cited by 96 · see citing works
Minzioni, P., Osellame, R., Sada, C., Zhao, S., Omenetto, F. G., Gylfason, K. B., Haraldsson, T., Zhang, Y., Özcan, A., Wax, A. P., et al. (2017) · Journal of Optics · doi:10.1088/2040-8986/aa783b · Cited by 94 · see citing works
Indel-correcting DNA barcodes for high-throughput sequencing
Hawkins, J. A., Jones, S. K., Finkelstein, I. J., & Press, W. H. (2018) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.1802640115 · Cited by 92 · see citing works
The Current Trends of Biosensors in Tissue Engineering
Li, Y., & Lee, I.-C. (2020) · Biosensors · doi:10.3390/bios10080088 · Cited by 53 · see citing works
Systems Approaches to Biology and Disease Enable Translational Systems Medicine
Hood, L., & Tian, Q. (2012) · Genomics Proteomics & Bioinformatics · doi:10.1016/j.gpb.2012.08.004 · Cited by 90 · see citing works
High-throughput spatiotemporal monitoring of single-cell secretions via plasmonic microwell arrays
Ansaryan, S., Liu, Y., Li, X., Economou, A. M., Eberhardt, C. S., Jandus, C., & Altug, H. (2023) · Nature Biomedical Engineering · doi:10.1038/s41551-023-01017-1 · Cited by 89 · see citing works
Abate‐Daga, D., Hanada, K., Davis, J. L., Yang, J. C.-H., Rosenberg, S. A., & Morgan, R. A. (2013) · Blood · doi:10.1182/blood-2013-04-495531 · Cited by 89 · see citing works
Single cells in confined volumes: microchambers and microdroplets
Hümmer, D., Kurth, F., Naredi‐Rainer, N., & Dittrich, P. S. (2015) · Lab on a Chip · doi:10.1039/c5lc01314c · Cited by 88 · see citing works
Su, Y., Shi, Q., & Wei, W. (2017) · PROTEOMICS · doi:10.1002/pmic.201600267 · Cited by 86 · see citing works
Self-citationThe Resistance Mechanisms of Lung Cancer Immunotherapy
Wang, F., Wang, S., & Zhou, Q. H. (2020) · Frontiers in Oncology · doi:10.3389/fonc.2020.568059 · Cited by 88 · see citing works
Eyer, K., Stratz, S., Kuhn, P., Küster, S. K., & Dittrich, P. S. (2013) · Analytical Chemistry · doi:10.1021/ac303628j · Cited by 88 · see citing works
Microfluidic single-cell technology in immunology and antibody screening
Seah, S., Hu, H., & Merten, C. A. (2017) · Molecular Aspects of Medicine · doi:10.1016/j.mam.2017.09.004 · Cited by 86 · see citing works
Evaluating Cellular Polyfunctionality with a Novel Polyfunctionality Index
Larsen, M., Sauce, D., Arnaud, L., Fastenackels, S., Appay, V., & Gorochov, G. (2012) · PLoS ONE · doi:10.1371/journal.pone.0042403 · Cited by 86 · see citing works
Microfluidics as an Enabling Technology for Personalized Cancer Therapy
Mathur, L., Ballinger, M., Utharala, R., & Merten, C. A. (2019) · Small · doi:10.1002/smll.201904321 · Cited by 83 · see citing works
Zhou, Y., Shao, N., de Castro, R. B., Zhang, P., Ma, Y., Liu, X., Huang, F., Wang, R.-F., & Qin, L. (2020) · Cell Reports · doi:10.1016/j.celrep.2020.107574 · Cited by 84 · see citing works
Advancing single-cell proteomics and metabolomics with microfluidic technologies
Liu, Y., Chen, X., Zhang, Y., & Liu, J. (2018) · The Analyst · doi:10.1039/c8an01503a · Cited by 80 · see citing works
Organs-on-chip monitoring: sensors and other strategies
Kilic, T., Navaee, F., Stradolini, F., Renaud, P., & Carrara, S. (2018) · Microphysiological Systems · doi:10.21037/mps.2018.01.01 · Cited by 96 · see citing works
Microfluidic Biochips for Single‐Cell Isolation and Single‐Cell Analysis of Multiomics and Exosomes
Wang, C., Qiu, J., Liu, M., Wang, Y., Yu, Y., Liu, H., Zhang, Y., & Han, L. G. (2024) · Advanced Science · doi:10.1002/advs.202401263 · Cited by 81 · see citing works
Torres, A. J., Hill, A. S., & Love, J. C. (2014) · Analytical Chemistry · doi:10.1021/ac4030297 · Cited by 77 · see citing works
Microfluidic and Paper-Based Devices for Disease Detection and Diagnostic Research
Campbell, J. M., Balhoff, J. B., Landwehr, G. M., Rahman, S. M., Vaithiyanathan, M., & Melvin, A. T. (2018) · International Journal of Molecular Sciences · doi:10.3390/ijms19092731 · Cited by 77 · see citing works
How single-cell immunology is benefiting from microfluidic technologies
Jammes, F., & Maerkl, S. J. (2020) · Microsystems & Nanoengineering · doi:10.1038/s41378-020-0140-8 · Cited by 73 · see citing works
Liu, Y., Matharu, Z., Howland, M. C., Revzin, A., & Simonian, A. L. (2012) · Analytical and Bioanalytical Chemistry · doi:10.1007/s00216-012-6149-6 · Cited by 75 · see citing works
Yu, J., Zhou, J., Sutherland, A., Wei, W., Shin, Y. S., Xue, M., & Heath, J. R. (2014) · Annual Review of Analytical Chemistry · doi:10.1146/annurev-anchem-071213-020323 · Cited by 72 · see citing works
Self-citationDeep Profiling of Cellular Heterogeneity by Emerging Single‐Cell Proteomic Technologies
Yang, L., George, J., & Wang, J. (2019) · PROTEOMICS · doi:10.1002/pmic.201900226 · Cited by 70 · see citing works
Fluorescent analysis of bioactive molecules in single cells based on microfluidic chips
Fan, Y., Dong, D., Li, Q., Si, H., Pei, H., Lu, L., & Tang, B. (2018) · Lab on a Chip · doi:10.1039/c7lc01333g · Cited by 71 · see citing works
Integrating Immunology and Microfluidics for Single Immune Cell Analysis
Sinha, N., Subedi, N., & Tel, J. (2018) · Frontiers in Immunology · doi:10.3389/fimmu.2018.02373 · Cited by 71 · see citing works
Microfluidics towards single cell resolution protein analysis
Chen, P., Chen, D., Li, S., Ou, X., & Liu, B. (2019) · TrAC Trends in Analytical Chemistry · doi:10.1016/j.trac.2019.06.022 · Cited by 70 · see citing works
Kim, J., Cho, H., Han, S., & Han, K.-H. (2016) · Analytical Chemistry · doi:10.1021/acs.analchem.6b00570 · Cited by 70 · see citing works
An integrated microfluidic platform for in situ cellular cytokine secretion immunophenotyping
Huang, N., Chen, W., Oh, B., Cornell, T. T., Shanley, T. P., Fu, J., & Kurabayashi, K. (2012) · Lab on a Chip · doi:10.1039/c2lc40619e · Cited by 69 · see citing works
Domain-swapped T cell receptors improve the safety of TCR gene therapy
Bethune, M. T., Gee, M. H., Bunse, M., Lee, M. S., Gschweng, E. H., Pagadala, M. S., Zhou, J., Cheng, D., Heath, J. R., Kohn, D. B., et al. (2016) · eLife · doi:10.7554/elife.19095 · Cited by 69 · see citing works
Self-citationSharafeldin, M., Chen, T., Özkaya, G. U., Choudhary, D., Molinolo, A., Gutkind, J. S., & Rusling, J. F. (2020) · Biosensors and Bioelectronics · doi:10.1016/j.bios.2020.112681 · Cited by 69 · see citing works
Chen, P., Huang, N., Chung, M. T., Cornell, T. T., & Kurabayashi, K. (2015) · Advanced Drug Delivery Reviews · doi:10.1016/j.addr.2015.09.005 · Cited by 68 · see citing works
- Authors
- Ma, C., Fan, R., Ahmad, H., Shi, Q., Comin-Anduix, B., Chodon, T., Koya, R. C., Liu, C.-C., Kwong, G. A., Radu, C. G., Ribas, A., & Heath, J. R.
- Affiliations
- California Institute of Technology (US)
- University of California, Los Angeles (US)
- Corresponding author
- Heath, James R
- ORCID
- Venue
- Nature Medicine
- Published
- 2011-05-22
- Volume
- 17
- Issue
- 6
- Pages
- 738-743
- ISSN
- 1078-8956 (ISSN-L)
- 1546-170X
- PMC
- PMC3681612
- OpenAlex
- W2166180044
- Semantic Scholar
- e7dd5bf60b18f6727615e597d7f083225c89ae27
- Cited by
- 441
- References
- 38
- Citations by year
- 2026: 6 · 2025: 11 · 2024: 19 · 2023: 31 · 2022: 28 · 2021: 18 · 2020: 29 · 2019: 42 · 2018: 47 · 2017: 41 · 2016: 28 · 2015: 27 · 2014: 35 · 2013: 40 · 2012: 29
- FWCI
- 14.82
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Licence
- www.springer.com/tdm (tdm)
- Locations
- Nature Medicine (publishedVersion)
- PubMed (publishedVersion)
- Europe PMC (PubMed Central) (acceptedVersion)
- PubMed Central (acceptedVersion)
- CaltechAUTHORS (California Institute of Technology) (submittedVersion)
- Funders
- California Institute for Regenerative Medicine
- Melanoma Research Alliance
- California Institute of Technology
- Jean Perkins Foundation
- National Institutes of Health (5u54ca119347-03, AI28697, CA16042, CA136759)
- National Cancer Institute (K99CA136759, CA-16 042, 5U54 CA119347, U54-CA119347, AI28697, CA 119347, P30CA016042)
- National Institute of Allergy and Infectious Diseases (AI-28 697, P30AI028697)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Advanced Biosensing Techniques and Applications
- Physical Sciences › Engineering › Biomedical Engineering › Microfluidic and Capillary Electrophoresis Applications
- Physical Sciences › Engineering › Biomedical Engineering › Microfluidic and Bio-sensing Technologies
- Keywords
- T cell functional heterogeneity, cytotoxic T lymphocytes, tumor antigen-specific T cells, inflammatory cytokines, microfluidic platform, immune monitoring
- MeSH terms
- Humans, Lipopolysaccharides, Macrophages, Neoplasms, Phenotype, T-Lymphocytes, T-Lymphocytes, Cytotoxic, Cytokines, Microchip Analytical Procedures, Lab-On-A-Chip Devices
About these checks
Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.
Statistics and arithmetic are recomputed, not judged.
Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.
What counts as a claim ›How we measure confidence ›How we check this ›
Are you an author, or do you see a problem with this page? Request a review
