Nanodroplets Based High Throughput Single-Cell PCR System

DESCRIPTION (provided by applicant): PCR System Chemical analysis of cellular contents, such as nucleic acids and proteins, is of significant interest to the biological, medical, and pharmaceutical communities. Detection of abnormal gene and protein in t he intracellular materials provide important clues for early disease diagnosis. The desire to perform precise molecular analysis on progressively smaller, pure cell populations has developed into the bourgeoning field of microg enomics. Unfortunately, investigators have tended to forgo the use of microgenomic technologies due to a lack of process control needed to ensure reliability of such studies. Key difficulties in microgenomic technologies are the difficult y in handling a minute amount of sample, inability to prepare and manipulate single cell, and inability to have high throughput capability. More importantly, it is extremely labor intensive and requires expensive equipment to isolate single cells and to perform amplification on each cell, so that the cell number that can be tested each time is very limited. Therefore, Maxwell Sensors Inc. proposes to develop a nanodrop based High Throughput Single Cell PCR (HTSC PCR) chip/system that will provide unprecedented information critical to the development of pathology, oncogenesis, and other processes of a desired target cell. The HTSC PCR system combines nanodroplet, microfluidics, and fluorescen t reverse transcription (RT) PCR technologies for performing single cell (SC) RT PCR in a quick, high throughput and cost effective fashion. This highly integrated platform is configured to precisely prepare and encapsulate a singl e cell in a nanodroplet, simultaneously perform hundreds or thousands of single cell PCR in nanoliter volumes, rapidly analyze the PCR results on chip, and open the road to single cell analyses of large populations for clinical diagnostics or biomedical research purpose. The combination of high fidelity manipulation of nanodroplets and the ability to perform nucleic acid amplification offers the possibility of developing powerful automated instruments.