NER: On-Chip Analysis of Individual Cells - Single-Molecule Counting of Rare Proteins

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0508531ZareThis proposal was received in response to Nanoscale Science and Engineering initiative, NSF 04-043, category NER.The applicant will combine single molecule detection with microfluidic manipulation to obtain absolute quantification of proteins within a single cell. The methods proposed will have the sensitivity required to determine the presence of proteins at low copy numbers, that is, one to a thousand copies per cell. Moreover, this goal can be achieved without (PCR-like) amplification, which for proteins is very difficult if not impossible. Currently, no such methods exist having these capabilities. The intellectual merit of this approach is that it provides the ultimate analytical tool for quantifying rare-copy-number proteins. It can detect as little as a single copy in a cell, and it provides an absolute count of the copies present.In addition, this one-year proposal offers the training of graduate students in state-of-the-art techniques that are at the forefront of nanoscale scientific research.This proposal will explore the development of new techniques, which, if successful, is expected to be widely adopted by others.