Single-cell analysis by a scanning thermal lens microscope with a microchip: direct monitoring of cytochrome c distribution during apoptosis process.
We developed a microsystem for cell experiments consisting of a scanning thermal lens microscope detection system and a cell culture microchip. The microchip system was good for liquid control in microspace, and this results in secure cell stimulation and coincident in vivo observation of the cell responses. The system could detect nonfluorescent biological substances with extremely high sensitivity without any labeling materials and had a high spatial resolution of approximately 1 microm. This system was applied to monitoring of cytochrome c distribution in a neuroblastoma-glioma hybrid cell cultured in the microflask (1 mm x 10 mm x 0.1 mm; 1 microL) fabricated in a glass microchip. Cytochrome c release from mitochondria to cytosol during the apoptosis process was successfully monitored with this system. The cytochrome c detected with this system was estimated to be approximately 10 zmol. We concluded that the system was suitable for measuring the distribution of chemical substances in a single cell because the microchip is good for liquid handling in microspace and the thermal lens microscope has high sensitivity and spatial resolution.
Tamaki
Eiichiro
E
Department of Applied Chemistry, School of Engineering, The University of Tokyo, Bunkyo, Japan.
Sato
Kiichi
K
Tokeshi
Manabu
M
Sato
Kae
K
Aihara
Makoto
M
Kitamori
Takehiko
T
D017209:Apoptosis; D003574:Cytochrome c Group / Q000032:analysis / Q000378:metabolism*; D003584:Cytological Techniques / Q000295:instrumentation*; D003593:Cytoplasm / Q000378:metabolism; D006801:Humans; D008853:Microscopy / Q000295:instrumentation* / Q000379:methods; D008904:Miniaturization; D008928:Mitochondria / Q000378:metabolism; D009447:Neuroblastoma / Q000378:metabolism / Q000473:pathology / Q000648:ultrastructure; D021381:Protein Transport
D003574:Cytochrome c Group