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Real-time single-cell imaging of protein secretion

by Nishikomori, Ryuta , Heike, Toshio , Harada, Yoshie , Mizuno, Jun , Izawa, Kazushi , Shirasaki, Yoshitaka , Suzuki, Nobutake , Yamagishi, Mai , Ohara, Osamu , Nakahara, Asahi , Shoji, Shuichi

in 631/1647/277 / 631/250/1933 / 631/553/2706 / 631/80/2373 / Bacterial proteins / Cell Membrane - metabolism / Cells, Cultured / Fluoroimmunoassay / Heterogeneity / Humanities and Social Sciences / Humans / IL-1β / Interleukin-1beta - metabolism / Interleukin-1beta - secretion / Interleukin-6 - metabolism / Interleukin-6 - secretion / Microscopy, Fluorescence / Molecular Imaging - methods / Monocytes / Monocytes - metabolism / multidisciplinary / Protein Transport / Science / Single-Cell Analysis - methods / T cell receptors

2014

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Real-time single-cell imaging of protein secretion

by Nishikomori, Ryuta , Heike, Toshio , Harada, Yoshie , Mizuno, Jun , Izawa, Kazushi , Shirasaki, Yoshitaka , Suzuki, Nobutake , Yamagishi, Mai , Ohara, Osamu , Nakahara, Asahi , Shoji, Shuichi

2014

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Real-time single-cell imaging of protein secretion

by Nishikomori, Ryuta , Heike, Toshio , Harada, Yoshie , Mizuno, Jun , Izawa, Kazushi , Shirasaki, Yoshitaka , Suzuki, Nobutake , Yamagishi, Mai , Ohara, Osamu , Nakahara, Asahi , Shoji, Shuichi

2014

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Journal Article

Real-time single-cell imaging of protein secretion

Nishikomori, Ryuta,

Heike, Toshio,

Harada, Yoshie,

Mizuno, Jun,

Izawa, Kazushi,

Shirasaki, Yoshitaka,

Suzuki, Nobutake,

Yamagishi, Mai,

Ohara, Osamu,

Nakahara, Asahi,

Shoji, Shuichi

2014

Overview

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.

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Publisher

Nature Publishing Group UK,Nature Publishing Group

Subject

/ Cell Membrane - metabolism

/ Cells, Cultured