Counting Low-Copy Number Proteins in a Single Cell

Bo Huang; Hongkai Wu; Devaki Bhaya; Arthur Grossman; Sebastien Granier; Brian K. Kobilka; Richard N. Zare

doi:10.1126/science.1133992

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Counting Low-Copy Number Proteins in a Single Cell

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Counting Low-Copy Number Proteins in a Single Cell

Bo Huang, Hongkai Wu, Devaki Bhaya, Arthur Grossman, Sebastien Granier, Brian K. Kobilka et Richard N. Zare

Science (American Association for the Advancement of Science), Vol.315(5808), pp.81-84

05/01/2007

DOI: https://doi.org/10.1126/science.1133992

PMID: 17204646

Résumé

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 (≈60%) counting of molecules in micrometer-sized channels. We used this microfluidic device to quantify β 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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Titre: Counting Low-Copy Number Proteins in a Single Cell
Créateurs - sans rôle: Bo Huang - Department of Chemistry, Stanford University, Stanford, CA 94305-5080, USA., Department of Plant Biology, Carnegie Institution, Stanford, CA 94305, USA., Department of Molecular and Cellular Physiology and Medicine, Stanford University, Stanford, CA 94305-5345, USA
Hongkai Wu - Department of Chemistry, Stanford University, Stanford, CA 94305-5080, USA., Department of Plant Biology, Carnegie Institution, Stanford, CA 94305, USA., Department of Molecular and Cellular Physiology and Medicine, Stanford University, Stanford, CA 94305-5345, USA
Devaki Bhaya - Department of Chemistry, Stanford University, Stanford, CA 94305-5080, USA., Department of Plant Biology, Carnegie Institution, Stanford, CA 94305, USA., Department of Molecular and Cellular Physiology and Medicine, Stanford University, Stanford, CA 94305-5345, USA
Arthur Grossman - Department of Chemistry, Stanford University, Stanford, CA 94305-5080, USA., Department of Plant Biology, Carnegie Institution, Stanford, CA 94305, USA., Department of Molecular and Cellular Physiology and Medicine, Stanford University, Stanford, CA 94305-5345, USA
Sebastien Granier - Department of Chemistry, Stanford University, Stanford, CA 94305-5080, USA., Department of Plant Biology, Carnegie Institution, Stanford, CA 94305, USA., Department of Molecular and Cellular Physiology and Medicine, Stanford University, Stanford, CA 94305-5345, USA
Brian K. Kobilka - Stanford University
Richard N. Zare - Department of Chemistry, Stanford University, Stanford, CA 94305-5080, USA., Department of Plant Biology, Carnegie Institution, Stanford, CA 94305, USA., Department of Molecular and Cellular Physiology and Medicine, Stanford University, Stanford, CA 94305-5345, USA
Détails de publication: Science (American Association for the Advancement of Science), Vol.315(5808), pp.81-84
Nombre de pages: 4
Identifiants: 99139573809311
Unité académique: Institut de Génomique Fonctionnelle - IGF
Langue: English
Type de ressource: Journal article