Microfluidic Devices for Single-cell Analysis
Yan Chen and Jiang F. Zhong
from: Lab-on-a-Chip Technology (Vol. 2): Biomolecular Separation and Analysis (Edited by: Keith E. Herold and Avraham Rasooly). Caister Academic Press, U.K. (2009)
Gene regulation is a continuous event. Differentiation/maturation of cells is orchestrated by sequentially expressing a series of genes within a cell after receiving signals from the microenvironment. Ultimately, the gene-gene interaction inside an individual cell determines the fate of that particular cell. Therefore, gene regulation should be studied at the single-cell level to understand how genes respond to environmental signals and subsequently produce proteins to regulate cellular activity. However, molecular biologists currently have limited ability to analyze the contents of a single-cell which typically are only several picoliters. Current devices and methods for biomedical research are designed for performing experiments at the microliter scale. Using those tools leads to significant material loss in single-cell analysis, and produces poor quality data.To overcome the technical barrier of single-cell analysis, we have designed and constructed microfluidic devices with computer-controlled microvalves and peristaltic pumps for biochemical analysis at the nanoliter scale. Carrying out biochemical reactions at the nanoliter scale significantly reduces material loss from small volumes of analyte such as a single-cell. Equipped with a thermal stage, our devices can extract mRNA from 32 or more individual cells and convert mRNA to cDNA within 3 hours with 5 fold higher efficiency than bulk assays. The microfluidic devices we developed can analyze single-cell contents precisely and produce reliable data. Those devices have the potential to transform single-cell analysis from a challenge and expensive task to a routine assay for molecular biologists read more ...