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Counting low-copy number proteins in a single cell
被引:312
作者:
Huang, Bo
Wu, Hongkai
Bhaya, Devaki
Grossman, Arthur
Granier, Sebastien
Kobilka, Brian K.
Zare, Richard N.
[1
]
机构:
[1] Stanford Univ, Dept Chem, Stanford, CA 94305 USA
[2] Carnegie Inst Washington, Dept Plant Biol, Stanford, CA 94305 USA
[3] Stanford Univ, Dept Mol & Cellular Physiol & Med, Stanford, CA 94305 USA
来源:
关键词:
D O I:
10.1126/science.1133992
中图分类号:
O [数理科学和化学];
P [天文学、地球科学];
Q [生物科学];
N [自然科学总论];
学科分类号:
07 ;
0710 ;
09 ;
摘要:
We have designed a microfluidic device in which we can manipulate, lyse, label, separate, and quantify the protein contents of a single cell using single-molecule fluorescence counting. Generic labeling of proteins is achieved through fluorescent-antibody binding. The use of cylindrical optics enables high-efficiency (approximate to 60%) counting of molecules in micrometer-sized channels. We used this microfluidic device to quantify beta(2) adrenergic receptors expressed in insect cells (SF9). We also analyzed phycobiliprotein contents in individual cyanobacterial cells ( Synechococcus sp. PCC 7942) and observed marked differences in the levels of specific complexes in cell populations that were grown under nitrogen-depleted conditions.
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页码:81 / 84
页数:4
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