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Single-Cell Quantitative Analysis of DNA Incorporation and Protein Expression in Microwells

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Abstract:

Conventional biochemical assays are performed via an averaging procedure with the lysate of a large number of target cells; however, the averaged data lose information regarding the heterogeneity of individual cells. For quantitative assay of single cells, it is necessary to isolate single cells, extract cellular components and detect extremely small amounts of molecules from the individual cells.

We developed new system combining a microfabricated lab-on-chip device and fluorescence correlation spectroscopy. The system features a simple protocol to isolate single-cells and detect small amounts of specific fluorescent molecules extracted from a single cell. Our results indicate that numbers of transfected DNA molecules were rather equal for each single cell; however, the expression rate of protein varied in each single cell.



Keywords: Cy5-DNA; Fluorescence correlation spectroscopy; PDMS chip; cell lysis; cellular heterogeneity; diffusion constant; expression rate; single cell isolation

Document Type: Research Article

DOI: http://dx.doi.org/10.2174/138920110790725393

Publication date: January 1, 2010

More about this publication?
  • Current Pharmaceutical Biotechnology aims to cover all the latest and outstanding developments in Pharmaceutical Biotechnology. Each issue of the journal contains a series of timely in-depth reviews written by leaders in the field covering a range of current topics in both pre-clinical and clinical areas of Pharmaceutical Biotechnology. Current Pharmaceutical Biotechnology is an essential journal for academic, clinical, government and pharmaceutical scientists who wish to be kept informed and up-to-date with the latest and most important developments.
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