Counting low-copy number proteins in a single cell

Huang, Bo; Wu, Hongkai; Bhaya, Devaki; Grossman, Arthur; Granier, Sebastien; Kobilka, Brian K.; Zare, Richard N.
2007
SCIENCE
DOI
10.1126/science.1133992
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.