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Publication : Use of the Fluidigm C1 platform for RNA sequencing of single mouse pancreatic islet cells.

First Author  Xin Y Year  2016
Journal  Proc Natl Acad Sci U S A Volume  113
Issue  12 Pages  3293-8
PubMed ID  26951663 Mgi Jnum  J:356580
Mgi Id  MGI:7762672 Doi  10.1073/pnas.1602306113
Citation  Xin Y, et al. (2016) Use of the Fluidigm C1 platform for RNA sequencing of single mouse pancreatic islet cells. Proc Natl Acad Sci U S A 113(12):3293-8
abstractText  This study provides an assessment of the Fluidigm C1 platform for RNA sequencing of single mouse pancreatic islet cells. The system combines microfluidic technology and nanoliter-scale reactions. We sequenced 622 cells, allowing identification of 341 islet cells with high-quality gene expression profiles. The cells clustered into populations of alpha-cells (5%), beta-cells (92%), delta-cells (1%), and pancreatic polypeptide cells (2%). We identified cell-type-specific transcription factors and pathways primarily involved in nutrient sensing and oxidation and cell signaling. Unexpectedly, 281 cells had to be removed from the analysis due to low viability, low sequencing quality, or contamination resulting in the detection of more than one islet hormone. Collectively, we provide a resource for identification of high-quality gene expression datasets to help expand insights into genes and pathways characterizing islet cell types. We reveal limitations in the C1 Fluidigm cell capture process resulting in contaminated cells with altered gene expression patterns. This calls for caution when interpreting single-cell transcriptomics data using the C1 Fluidigm system.
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