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 language | English |
|---|---|
| Article number | 4736 |
| Journal | Scientific Reports |
| Volume | 4 |
| DOIs | |
| Publication status | Published - 2014 Apr 22 |
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ASJC Scopus subject areas
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Cite this
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 journal › Article
}
TY - JOUR
T1 - Real-time single-cell imaging of protein secretion
AU - Shirasaki, Yoshitaka
AU - Yamagishi, Mai
AU - Suzuki, Nobutake
AU - Izawa, Kazushi
AU - Nakahara, Asahi
AU - Mizuno, Jun
AU - Shoji, Shuichi
AU - Heike, Toshio
AU - Harada, Yoshie
AU - Nishikomori, Ryuta
AU - Ohara, Osamu
PY - 2014/4/22
Y1 - 2014/4/22
N2 - 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.
AB - 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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UR - http://www.scopus.com/inward/citedby.url?scp=84899063207&partnerID=8YFLogxK
U2 - 10.1038/srep04736
DO - 10.1038/srep04736
M3 - Article
C2 - 24751898
AN - SCOPUS:84899063207
VL - 4
JO - Scientific Reports
JF - Scientific Reports
SN - 2045-2322
M1 - 4736
ER -