Review of:A Nanoliter-Scale Nucleic Acid

Processor with Parallel ArchitectureTiffany Guo

October 10, 200820.309

J. W. Hong, et al. "A nanoliter-scale nucleic acid processor with parallel architecture," Nature Biotech. 22(4): pp. 435-439 (2004)

Single Cell Nucleic Acid Isolation

• Study presence (through DNA) or expression (through RNA) of specific genes– Identify cells of specific genotype for

experimentation– Offer clues to differential phenotypes under

specific conditions

• Study transcriptional variation between cells of same genotype

How It Works:

The Components

How it works:

Mixing

How it works: DNA Isolation by Affinity Column

How it works:

Elution

Parallel Architecture

Microfluidics

• Process requires minimal numbers of cells– Eliminate overnight culturing

steps– Study expression on the

single cell level– Study expression of slow

growing cells more easily

Application: Variable Expression of Motility Genes in Tumor Cells

W. Wang, et al. “Tumor cells caught in the act of invading: their strategy for enhanced cell motility,” TRENDS in Cell Bio. 15(3): pp. 138-145 (2005)

“There is, however, the possibility that these expression patterns reflect the noninvasive cells that make up the bulk of the tumor mass and that the expression signature of invasive tumor cells – arguably the motile population essential for metasasis – might be masked.”

“If a specific combination of gene expression patterns can be correlated with metastic ability, however, then such analyses can be predictive of metastatic outcome…”

Future Directions: Micro PCRE.A. Ottesen, et al. “Microfluidic Digital PCR Enables Multigene Analysis of Individual Environmental Bacteria, Science, 314: pp. 1464-1467 (2006)

QUESTIONS?