Stochastic particle barcoding for single-cell tracking and multiparametric analysis.

This study presents stochastic particle barcoding (SPB), a method for tracking cell identity across bioanalytical platforms. In this approach, single cells or small collections of cells are co-encapsulated within an enzymatically-degradable hydrogel block along with a random collection of fluorescent beads, whose number, color, and position encode the identity of the cell, enabling samples to be transferred in bulk between single-cell assay platforms without losing the identity of individual cells. The application of SPB is demonstrated for transferring cells from a subnanoliter protein secretion/phenotyping array platform into a microtiter plate, with re-identification accuracies in the plate assay of 96±2%. Encapsulated cells are recovered by digesting the hydrogel, allowing subsequent genotyping and phenotyping of cell lysates. Finally, a model scaling is developed to illustrate how different parameters affect the accuracy of SPB and to motivate scaling of the method to thousands of unique blocks.

[1]  J. Christopher Love,et al.  Single-Cell Analysis Reveals Isotype-Specific Autoreactive B Cell Repertoires in Sjögren’s Syndrome , 2013, PloS one.

[2]  Stephen R. Quake,et al.  Genome-wide Single-Cell Analysis of Recombination Activity and De Novo Mutation Rates in Human Sperm , 2012, Cell.

[3]  O. Urakawa,et al.  Small - , 2007 .

[4]  L. Vermeulen,et al.  Cancer heterogeneity—a multifaceted view , 2013, EMBO reports.

[5]  P. Taylor,et al.  Macrophage heterogeneity and acute inflammation , 2011, European journal of immunology.

[6]  M P Murtaugh,et al.  Regulation of hypoxanthine phosphoribosyltransferase, glyceraldehyde-3-phosphate dehydrogenase and beta-actin mRNA expression in porcine immune cells and tissues. , 1998, Animal biotechnology.

[7]  Samuel Aparicio,et al.  High-throughput microfluidic single-cell RT-qPCR , 2011, Proceedings of the National Academy of Sciences.

[8]  J. C. Love,et al.  Screening individual hybridomas by microengraving to discover monoclonal antibodies , 2009, Nature Protocols.

[9]  Sailing He,et al.  Rapid Fabrication of Complex 3D Extracellular Microenvironments by Dynamic Optical Projection Stereolithography , 2012, Advanced materials.

[10]  P. Sorger,et al.  Non-genetic origins of cell-to-cell variability in TRAIL-induced apoptosis , 2009, Nature.

[11]  Helene Andersson Svahn,et al.  Droplet microfluidics--a tool for single-cell analysis. , 2012, Angewandte Chemie.

[12]  Chad A Mirkin,et al.  A fluorophore-based bio-barcode amplification assay for proteins. , 2006, Small.

[13]  D K Wood,et al.  A feasible approach to all-electronic digital labeling and readout for cell identification. , 2007, Lab on a chip.

[14]  P. Swain,et al.  Stochastic Gene Expression in a Single Cell , 2002, Science.

[15]  Rong Fan,et al.  A Clinical Microchip for Evaluation of Single Immune Cells Reveals High Functional Heterogeneity in Phenotypically Similar T Cells Nih Public Access Author Manuscript Design Rationale and Detection Limit of the Scbc Online Methods Microchip Fabrication On-chip Secretion Profiling Supplementary Mater , 2022 .

[16]  Jaume Esteve,et al.  Intracellular polysilicon barcodes for cell tracking. , 2009, Small.

[17]  J. Gershoni,et al.  Profiling the IgOme: Meeting the challenge , 2013, FEBS Letters.

[18]  P. Chattopadhyay,et al.  Seventeen-colour flow cytometry: unravelling the immune system , 2004, Nature Reviews Immunology.

[19]  J. C. Love,et al.  A microengraving method for rapid selection of single cells producing antigen-specific antibodies , 2006, Nature Biotechnology.

[20]  Daniel C Douek,et al.  Bias in the αβ T‐cell repertoire: implications for disease pathogenesis and vaccination , 2011, Immunology and cell biology.

[21]  J. Mattick,et al.  Genome research , 1990, Nature.

[22]  Min-Gon Kim,et al.  Addressable micropatterning of multiple proteins and cells by microscope projection photolithography based on a protein friendly photoresist. , 2010, Langmuir : the ACS journal of surfaces and colloids.

[23]  Junsang Doh,et al.  Photogenerated polyelectrolyte bilayers from an aqueous-processible photoresist for multicomponent protein patterning. , 2004, Journal of the American Chemical Society.

[24]  G. Church,et al.  Barcoding cells using cell-surface programmable DNA-binding domains , 2013, Nature Methods.

[25]  D. Hayes,et al.  Circulating tumour cells: insights into tumour heterogeneity , 2013, Journal of internal medicine.

[26]  Shahar Alon,et al.  Barcoding bias in high-throughput multiplex sequencing of miRNA. , 2011, Genome research.

[27]  Ikuo Fujii,et al.  An automated system for high-throughput single cell-based breeding , 2013, Scientific Reports.

[28]  Navin Varadarajan,et al.  Rapid, efficient functional characterization and recovery of HIV-specific human CD8+ T cells using microengraving , 2012, Proceedings of the National Academy of Sciences.

[29]  T. Golub,et al.  A method for high-throughput gene expression signature analysis , 2006, Genome Biology.

[30]  Dhananjay Dendukuri,et al.  Continuous-flow lithography for high-throughput microparticle synthesis , 2006, Nature materials.

[31]  Alex Rhee,et al.  Facile and rapid one-step mass preparation of quantum-dot barcodes. , 2008, Angewandte Chemie.

[32]  Boris E. Burakov,et al.  Advanced Materials , 2019, Springer Proceedings in Physics.

[33]  Mario Roederer,et al.  Single-cell technologies for monitoring immune systems , 2014, Nature Immunology.

[34]  Niels W. Hanson,et al.  A programmable droplet-based microfluidic device applied to multiparameter analysis of single microbes and microbial communities , 2012, Proceedings of the National Academy of Sciences.

[35]  N. Perrimon,et al.  Droplet microfluidic technology for single-cell high-throughput screening , 2009, Proceedings of the National Academy of Sciences.

[36]  J. Christopher Love,et al.  Generation and Screening of Pichia pastoris Strains with Enhanced Protein Production by Use of Microengraving , 2011, Applied and Environmental Microbiology.

[37]  A. Schmid,et al.  Single-cell analysis in biotechnology, systems biology, and biocatalysis. , 2012, Annual review of chemical and biomolecular engineering.

[38]  D. Irvine,et al.  Composition-tunable properties of amphiphilic comb copolymers containing protected methacrylic acid groups for multicomponent protein patterning. , 2006, Langmuir : the ACS journal of surfaces and colloids.

[39]  J Christopher Love,et al.  Cellular barcodes for efficiently profiling single-cell secretory responses by microengraving. , 2012, Analytical chemistry.

[40]  Stephen R Quake,et al.  Whole-genome molecular haplotyping of single cells , 2011, Nature Biotechnology.

[41]  E. Hall,et al.  The nature of biotechnology. , 1988, Journal of biomedical engineering.

[42]  References , 1971 .

[43]  Genbank,et al.  APPLIED AND ENVIRONMENTAL MICROBIOLOGY , 2008, Applied and Environmental Microbiology.

[44]  O. Bagasra,et al.  Proceedings of the National Academy of Sciences , 1914, Science.

[45]  J. West,et al.  Proteolytically Degradable Hydrogels with a Fluorogenic Substrate for Studies of Cellular Proteolytic Activity and Migration , 2005, Biotechnology progress.

[46]  Melinda Fitzgerald,et al.  Immunol. Cell Biol. , 1995 .

[47]  J. C. Love,et al.  Systematic Single-Cell Analysis of Pichia pastoris Reveals Secretory Capacity Limits Productivity , 2012, PloS one.

[48]  R. Williams,et al.  Journal of American Chemical Society , 1979 .

[49]  J. Christopher Love,et al.  Integrated process design for single‐cell analytical technologies , 2010 .

[50]  A Leonard,et al.  Internal medicine. , 1980, Journal of medical education.

[51]  R. Rosenfeld Nature , 2009, Otolaryngology--head and neck surgery : official journal of American Academy of Otolaryngology-Head and Neck Surgery.

[52]  J. C. Love,et al.  Profiling antibody responses by multiparametric analysis of primary B cells , 2008, Proceedings of the National Academy of Sciences.

[53]  Garry P Nolan,et al.  Fluorescent cell barcoding in flow cytometry allows high-throughput drug screening and signaling profiling , 2006, Nature Methods.

[54]  S. Arii,et al.  Phenotype and functional identity of GM-CSF-independent dendritic cells generated by long-term propagation of DC progenitor cells in bone marrow cells and skin Langerhans cells. , 2005 .

[55]  J Christopher Love,et al.  Development and optimization of a process for automated recovery of single cells identified by microengraving , 2010, Biotechnology progress.