title: Vertical flow array chips reliably identify cell types from single-cell mRNA sequencing experiments.
abstract: Single-cell mRNA sequencing offers an unbiased approach to dissecting cell types as functional units in multicellular tissues. However, highly reliable cell typing based on single-cell gene expression analysis remains challenging because of the lack of methods for efficient sample preparation for high-throughput sequencing and evaluating the statistical reliability of the acquired cell types. Here, we present a highly efficient nucleic reaction chip (a vertical flow array chip (VFAC)) that uses porous materials to reduce measurement noise and improve throughput without a substantial increase in reagent. We also present a probabilistic evaluation method for cell typing depending on the amount of measurement noise. Applying the VFACs to 2580 monocytes provides 1967 single-cell expressions for 47 genes, including low-expression genes such as transcription factors. The statistical method can distinguish two cell types with probabilistic quality values, with the measurement noise level being considered for the first time. This approach enables the identification of various sub-types of cells in tissues and provides a foundation for subsequent analyses.
authors.affiliation: Hitachi, Ltd., Research &Development Group 1-280, Higashi-koigakubo, kokubunji-shi, Tokyo, Japan.
authors.forename: Masataka
authors.initials: M
authors.lastname: Shirai
authors.affiliation: Hitachi, Ltd., Research &Development Group 1-280, Higashi-koigakubo, kokubunji-shi, Tokyo, Japan.
authors.forename: Koji
authors.initials: K
authors.lastname: Arikawa
authors.affiliation: Hitachi, Ltd., Research &Development Group 1-280, Higashi-koigakubo, kokubunji-shi, Tokyo, Japan.
authors.forename: Kiyomi
authors.initials: K
authors.lastname: Taniguchi
authors.affiliation: Hitachi, Ltd., Research &Development Group 1-280, Higashi-koigakubo, kokubunji-shi, Tokyo, Japan.
authors.forename: Maiko
authors.initials: M
authors.lastname: Tanabe
authors.affiliation: Hitachi, Ltd., Research &Development Group 1-280, Higashi-koigakubo, kokubunji-shi, Tokyo, Japan.
authors.forename: Tomoyuki
authors.initials: T
authors.lastname: Sakai
mesh_terms: D003584:Cytological Techniques / Q000379:methods*; D020869:Gene Expression Profiling / Q000379:methods; D006801:Humans; D046228:Microarray Analysis / Q000379:methods; D009000:Monocytes / Q000145:classification*; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D012333:RNA, Messenger / Q000235:genetics*; D017423:Sequence Analysis, RNA / Q000379:methods*; D059010:Single-Cell Analysis / Q000379:methods*; D000074084:THP-1 Cells
chemical_list: D012333:RNA, Messenger