The cells are deposited into nanopen chambers where they proliferate and can be tested using multiple assays. The Beacon system developed by Berkeley Lights combines optics and nanofluidics for processing and deep profiling of single cells. In this study, single B-cells from rat, mouse and human were sequenced using the 10x Genomics Chromium system to obtain paired full-length antibody variable regions, demonstrating that the method is incredibly effective for rapid antibody discovery. Immune profiling of full-length V(D)J sequences for paired B-cell or T-cell receptors is also possible, as shown in Goldstein et al. This technology allows transcriptome sequencing of thousands of individual cells, making it possible to study immune cell populations and identify cellular subtypes in the sample. 10x Genomics Chromium workflows, from sample to barcoded cDNA. All generated cDNA from a single cell share common 10x barcodes.įigure 1. ![]() The cell inside a GEM undergoes lysis and reverse transcription to generate 10x barcoded cDNA, which is then transferred to a tube for amplification, library construction and parallel sequencing (Fig. A gel bead in emulsion (GEM) with unique 10x barcodes, sequencing adapters and primers is used to encapsulate each cell. The 10x Genomics Chromium system makes use of droplet microfluidics to sequence mRNA of single cells. Furthermore, we discuss some of the challenges that arise when analyzing single-cell data, and we feature how the PipeBio Bioinformatics platform supports and processes data exported from single-cell sequencing platforms. Here, we highlight two of the main single-cell sequencing technologies based on microfluidics available nowadays: the 10x Genomics Chromium system and the Beacon system. Microfluidic-based assays are able to dissect major immune populations in unprecedented speed, retrieve antibody chain pairs accurately and recover the processed sequences for downstream analysis. The isolated RNA is labeled with cell-specific barcodes, so the different sequences can be traced back to the original cell after being sequenced. These methods use special miniaturized platforms which enable the compartmentalization of single cells in either wells or droplets. In the past years, several transcriptomic technologies based on microfluidic assays have emerged, enabling accurate and high-throughput sequencing of single cells. The sorted cells are subsequently sequenced using Sanger sequencing or NGS. For RNA sequencing of the whole transcriptome, Fluorescence-activated Cell Sorting (FACS) has been used to sort distinct immune populations and to isolate single ASC based on cell surface markers or staining. ![]() qPCR can measure multiple genes simultaneously from hundreds of cells, but the process is tedious and lacks whole-transcriptome analysis. One of the first technologies used to study gene expression in single cells was qPCR. Ultimately, these approaches accelerate the development of antibody-based therapeutics by directly isolating and sequencing antibody-secreting cells (ASCs) to select antigen-specific antibodies. Single-cell methods offer a deep insight into the complex biology of the immune system and enable the precise study of cell secretion, migration, cell-to-cell interaction and mRNA sequencing. With the current advancements in antibody drug discovery, there is an increasing demand for sequencing technologies to provide efficient tools able to characterize single immunological cells.
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