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

Research output: Contribution to journalArticle

49 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

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Interleukin-1
Proteins
Immunoassay
Monocytes
Microscopy
Membranes
single cell proteins
Population

ASJC Scopus subject areas

  • General

Cite this

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

Real-time single-cell imaging of protein secretion. / Shirasaki, Yoshitaka; Yamagishi, Mai; Suzuki, Nobutake; Izawa, Kazushi; Nakahara, Asahi; Mizuno, Jun; Shoji, Shuichi; Heike, Toshio; Harada, Yoshie; Nishikomori, Ryuta; Ohara, Osamu.

In: Scientific Reports, Vol. 4, 4736, 22.04.2014.

Research output: Contribution to journalArticle

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, vol. 4, 4736. https://doi.org/10.1038/srep04736
Shirasaki Y, Yamagishi M, Suzuki N, Izawa K, Nakahara A, Mizuno J et al. Real-time single-cell imaging of protein secretion. Scientific Reports. 2014 Apr 22;4. 4736. https://doi.org/10.1038/srep04736
Shirasaki, Yoshitaka ; Yamagishi, Mai ; Suzuki, Nobutake ; Izawa, Kazushi ; Nakahara, Asahi ; Mizuno, Jun ; Shoji, Shuichi ; Heike, Toshio ; Harada, Yoshie ; Nishikomori, Ryuta ; Ohara, Osamu. / Real-time single-cell imaging of protein secretion. In: Scientific Reports. 2014 ; Vol. 4.
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