An evanescent approach for mitochondrial function assay of living cells.
Mitochondria are now known to function physiologically not only in the production of ATP as the major cellular energy source, but also in the regulation of intracellular signaling, in, for example, stress-induced apoptosis and buffering of cytosolic calcium. It should be noted, when interpreting mitochondrial studies in situ, that mitochondria within cells show heterogeneity in both function and location. We applied both conventional epifluorescence microscopy (EPIFM) and total-internal-reflection fluorescence microscopy (TIRFM) in this study. Image data taken from TIRFM are excellent and markedly different from those taken from EPIFM. We further investigated the physiological variations of mitochondrial functions using an EPIFM/TIRFM dual-imaging system. This system permits further analysis of functions of mitochondria and other organelles with more precision than is possible using a traditional platform.
Yang
De-Ming
DM
Department of Medical Research and Education, Taipei Veterans General Hospital, Taipei, Taiwan. dmyang@vghtpe.gov.tw
Huang
Chien-Chang
CC
Kao
Lung-Sen
LS
Lin
Chung-Chih
CC
Chi
Chin-Wen
CW
Lin
Hsia Yu
HY
Tsai
Din Ping
DP
Lee
Chen-Hsen
CH
Chiou
Arthur
A
D000818:Animals; D002118:Calcium / Q000378:metabolism*; D007089:Image Enhancement / Q000379:methods*; D018613:Microscopy, Confocal / Q000379:methods*; D008856:Microscopy, Fluorescence / Q000379:methods*; D018715:Microscopy, Video / Q000379:methods*; D008928:Mitochondria / Q000502:physiology* / Q000648:ultrastructure*; D016716:PC12 Cells; D051381:Rats
D002118:Calcium