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Science 315 (5808): 81-84

Copyright © 2007 by the American Association for the Advancement of Science

Counting Low-Copy Number Proteins in a Single Cell

Bo Huang,1* Hongkai Wu,1{dagger} Devaki Bhaya,2 Arthur Grossman,2 Sebastien Granier,3 Brian K. Kobilka,3 Richard N. Zare1{ddagger}

Abstract: 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 ({approx}60%) counting of molecules in micrometer-sized channels. We used this microfluidic device to quantify ß2 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.

1 Department of Chemistry, Stanford University, Stanford, CA 94305-5080, USA.
2 Department of Plant Biology, Carnegie Institution, Stanford, CA 94305, USA.
3 Department of Molecular and Cellular Physiology and Medicine, Stanford University, Stanford, CA 94305-5345, USA.

* Present address: Department of Chemistry and Chemical Biology, Harvard University, Cambridge, MA 02138, USA.

{dagger} Present address: Department of Chemistry, Tsinghua University, Beijing 100084, China.

{ddagger} To whom correspondence should be addressed. E-mail: zare{at}stanford.edu


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