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
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.
引用
收藏
页码:81 / 84
页数:4
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