Real-time single-cell imaging of protein secretion

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

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54 Citations (Scopus)

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.

Original languageEnglish
Article number4736
JournalScientific reports
Volume4
DOIs
Publication statusPublished - 2014 Apr 22

ASJC Scopus subject areas

  • General

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    Shirasaki, Y., Yamagishi, M., Suzuki, N., Izawa, K., Nakahara, A., Mizuno, J., Shoji, S., Heike, T., Harada, Y., Nishikomori, R., & Ohara, O. (2014). Real-time single-cell imaging of protein secretion. Scientific reports, 4, [4736]. https://doi.org/10.1038/srep04736