Title :
Single cell real time secretion assay using amorphous fluoropolymer microwell array
Author :
Shirasaki, Y. ; Nakahara, A. ; Shimura, N. ; Yamagishi, M. ; Mizuno, J. ; Ohara, O. ; Shoji, S.
Author_Institution :
RCAI, RIKEN, Wako, Japan
Abstract :
Real time assay of protein secretion from living single cells was realized by coupling total internal reflection fluorescence (TIRF) microscopy observation and an optically optimized microwell array device. This system was based on a sandwich immunosorbent assay. Individual cells were cultured on an inverted microscope in microwell arrays fabricated of an amorphous fluoropolymer, CYTOP, to be monitored by TIRF imaging. CYTOP have no interference for the TIRF imaging because the refractive index of CYTOP is almost same as that of water. Using this system, we could succeed in real time secretion assay of lipopolysaccharide stimulated mast cells and the secretion time course of IL-1β from a single MC/9 cell was obtained successfully.
Keywords :
bioMEMS; biochemistry; cellular biophysics; fluorescence; microfabrication; molecular biophysics; noncrystalline structure; optical microscopy; polymers; proteins; refractive index; IL-1β; amorphous fluoropolymer; amorphous fluoropolymer microwell array; coupling total internal reflection fluorescence microscopy; inverted microscope; lipopolysaccharide stimulated mast cells; optically optimized microwell array device; protein secretion; real time secretion assay; refractive index; sandwich immunosorbent assay; secretion time course; single MC/9 cell; single cell real time secretion assay; Biomedical monitoring; Immune system; Materials; Monitoring; Optical imaging; Optical waveguides; CYTOP; TIRF; secretion assay; single cell;
Conference_Titel :
Solid-State Sensors, Actuators and Microsystems Conference (TRANSDUCERS), 2011 16th International
Conference_Location :
Beijing
Print_ISBN :
978-1-4577-0157-3
DOI :
10.1109/TRANSDUCERS.2011.5969330
