High-throughput multiparameter analysis of individual mitochondria.
Mitochondria are one of the most important organelles responsible for cellular energy metabolism and apoptosis regulation. However, single-mitochondrion analysis is challenging, because of their small sizes and the low content of organelle constituents. Here, we report the development of a sensitive and versatile platform for high-throughput multiparameter analysis of individual mitochondria. Employing specific fluorescent staining with a laboratory-built high-sensitivity flow cytometer (HSFCM), we demonstrate the simultaneous detection of side scatter, cardiolipin, and mitochondria DNA (mtDNA) of a single mitochondrion. Simultaneous measurements of side scatter, porin, and cytochrome c of individual mitochondria are reported for the first time. Correlation analysis among multiple attributes on an organelle-by-organelle basis could provide a more definitive assessment of the purity, structure integrity, and apoptosis-related proteins of isolated mitochondria than bulk measurement. This work represents a significant advancement in single-mitochondrion analysis. We believe that the HSFCM holds great potential for studying apoptotic signal transduction pathways at the single-mitochondrion level.
The Key Laboratory of Analytical Science, Department of Chemical Biology, College of Chemistry and Chemical Engineering, Xiamen University, Xiamen, Fujian 361005, People's Republic of China.
Shuyue
S
Zhang
Shaobin
S
Zhu
Lingling
L
Yang
Yan
Y
Zheng
Min
M
Gao
Shuo
S
Wang
Jin-zhang
JZ
Zeng
Xiaomei
X
Yan
D002308:Cardiolipins / Q000032:analysis; D045304:Cytochromes c / Q000032:analysis; D004272:DNA, Mitochondrial / Q000032:analysis; D005434:Flow Cytometry; D016650:Fluorescein-5-isothiocyanate / Q000737:chemistry; D031541:Fluorescence Resonance Energy Transfer; D006367:HeLa Cells; D006801:Humans; D008928:Mitochondria / Q000378:metabolism*; D053758:Nanoparticles / Q000737:chemistry; D012822:Silicon Dioxide / Q000737:chemistry
D002308:Cardiolipins; D004272:DNA, Mitochondrial; D012822:Silicon Dioxide; D045304:Cytochromes c; D016650:Fluorescein-5-isothiocyanate