title: Real-time single-cell imaging of protein secretion.
abstract: Protein secretion, a key intercellular event for transducing cellular signals, is thought to be strictly regulated. However, secretion dynamics at the single-cell level have not yet been clarified because intercellular heterogeneity results in an averaging response from the bulk cell population. To address this issue, we developed a novel assay platform for real-time imaging of protein secretion at single-cell resolution by a sandwich immunoassay monitored by total internal reflection microscopy in sub-nanolitre-sized microwell arrays. Real-time secretion imaging on the platform at 1-min time intervals allowed successful detection of the heterogeneous onset time of nonclassical IL-1β secretion from monocytes after external stimulation. The platform also helped in elucidating the chronological relationship between loss of membrane integrity and IL-1β secretion. The study results indicate that this unique monitoring platform will serve as a new and powerful tool for analysing protein secretion dynamics with simultaneous monitoring of intracellular events by live-cell imaging.
authors.affiliation: RIKEN Center for Integrative Medical Sciences (IMS-RCAI), 1-7-22 Suehiro-cho Tsurumi-ku, Yokohama, Kanagawa 230-0045, Japan.
authors.forename: Yoshitaka
authors.initials: Y
authors.lastname: Shirasaki
authors.affiliation: RIKEN Center for Integrative Medical Sciences (IMS-RCAI), 1-7-22 Suehiro-cho Tsurumi-ku, Yokohama, Kanagawa 230-0045, Japan.
authors.forename: Mai
authors.initials: M
authors.lastname: Yamagishi
authors.affiliation: RIKEN Center for Integrative Medical Sciences (IMS-RCAI), 1-7-22 Suehiro-cho Tsurumi-ku, Yokohama, Kanagawa 230-0045, Japan.
authors.forename: Nobutake
authors.initials: N
authors.lastname: Suzuki
authors.affiliation: Department of Pediatrics, Kyoto University Graduate School of Medicine, 54 Shogoin-Kawahara-cho Sakyo-ku, Kyoto 606-8507, Japan.
authors.forename: Kazushi
authors.initials: K
authors.lastname: Izawa
authors.affiliation: Faculty of Science and Engineering, Waseda University, Okubo 3-4-1, Shinjuku, Tokyo 169-8555, Japan.
authors.forename: Asahi
authors.initials: A
authors.lastname: Nakahara
authors.affiliation: Faculty of Science and Engineering, Waseda University, Okubo 3-4-1, Shinjuku, Tokyo 169-8555, Japan.
authors.forename: Jun
authors.initials: J
authors.lastname: Mizuno
authors.affiliation: Faculty of Science and Engineering, Waseda University, Okubo 3-4-1, Shinjuku, Tokyo 169-8555, Japan.
authors.forename: Shuichi
authors.initials: S
authors.lastname: Shoji
authors.affiliation: Department of Pediatrics, Kyoto University Graduate School of Medicine, 54 Shogoin-Kawahara-cho Sakyo-ku, Kyoto 606-8507, Japan.
authors.forename: Toshio
authors.initials: T
authors.lastname: Heike
authors.affiliation: Institute for Integrated Cell-Material Sciences (WPI-iCeMS), Kyoto University Graduate School of Biostudies, Yoshida-Honmachi, Sakyo-ku, Kyoto 606-8501, Japan.
authors.forename: Yoshie
authors.initials: Y
authors.lastname: Harada
authors.affiliation: Department of Pediatrics, Kyoto University Graduate School of Medicine, 54 Shogoin-Kawahara-cho Sakyo-ku, Kyoto 606-8507, Japan.
authors.forename: Ryuta
authors.initials: R
authors.lastname: Nishikomori
authors.affiliation: 1] RIKEN Center for Integrative Medical Sciences (IMS-RCAI), 1-7-22 Suehiro-cho Tsurumi-ku, Yokohama, Kanagawa 230-0045, Japan [2] Department of Human Genome Research, Kazusa DNA Research Institute, 2-6-7 Kazusa-Kamatari, Kisarazu, Chiba 292-0818, Japan.
authors.forename: Osamu
authors.initials: O
authors.lastname: Ohara
mesh_terms: D002462:Cell Membrane / Q000378:metabolism; D002478:Cells, Cultured; D015200:Fluoroimmunoassay; D006801:Humans; D053583:Interleukin-1beta / Q000378:metabolism; D015850:Interleukin-6 / Q000378:metabolism; D008856:Microscopy, Fluorescence; D057054:Molecular Imaging / Q000379:methods*; D009000:Monocytes / Q000378:metabolism; D021381:Protein Transport; D059010:Single-Cell Analysis / Q000379:methods*
chemical_list: D053583:Interleukin-1beta; D015850:Interleukin-6
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