RRC ID 45463
Author Kurokawa H, Ito H, Inoue M, Tabata K, Sato Y, Yamagata K, Kizaka-Kondoh S, Kadonosono T, Yano S, Inoue M, Kamachi T.
Title High resolution imaging of intracellular oxygen concentration by phosphorescence lifetime.
Journal Sci Rep
Abstract Optical methods using phosphorescence quenching by oxygen are suitable for sequential monitoring and non-invasive measurements for oxygen concentration (OC) imaging within cells. Phosphorescence intensity measurement is widely used with phosphorescent dyes. These dyes are ubiquitously but heterogeneously distributed inside the whole cell. The distribution of phosphorescent dye is a major disadvantage in phosphorescence intensity measurement. We established OC imaging system for a single cell using phosphorescence lifetime and a laser scanning confocal microscope. This system had improved spatial resolution and reduced the measurement time with the high repetition rate of the laser. By the combination of ubiquitously distributed phosphorescent dye with this lifetime imaging microscope, we can visualize the OC inside the whole cell and spheroid. This system uses reversible phosphorescence quenching by oxygen, so it can measure successive OC changes from normoxia to anoxia. Lower regions of OC inside the cell colocalized with mitochondria. The time-dependent OC change in an insulin-producing cell line MIN6 by the glucose stimulation was successfully visualized. Assessing the detailed distribution and dynamics of OC inside cells achieved by the presented system will be useful to understanding a physiological and pathological oxygen metabolism.
Volume 5
Pages 10657
Published 2015-6-12
DOI 10.1038/srep10657
PII srep10657
PMID 26065366
PMC PMC4464287
MeSH Animals Cell Hypoxia / physiology Cell Line, Tumor Humans Mice Molecular Imaging / methods* Oxygen / metabolism*
IF 3.998
Times Cited 63
WOS Category BIOCHEMISTRY & MOLECULAR BIOLOGY
Resource
Human and Animal Cells Colon-26(RCB2657) MKN45(RCB1001)