Single embryo gene expression

The new Research Spotlight paper describes how the Stanford scientists focused their research efforts on investigating the role of Oct4, a pluripotency regulator in embryonic stem cells. To find out what the Oct4 pluripotency regulator does at the one- to two-cell stage, the researchers undertook Gene Expression Analysis on 42 different genes using Fluidigm's BioMark™ System for Genetic Analysis and then used Fluidigm's 48.48 dynamic array integrated fluidic circuit to validate differential gene expression between knockdown and control samples.

The Stanford University researchers found that Oct4, the master regulator of embryonic stem cell pluripotency, also has critical functions during reprogramming of the early mammalian embryo. The researchers acknowledge how the consistent, high quality single embryo data has allowed them to identify genes that are consistently differentially regulated and to find rare outlier embryos expressing unique transcriptomes.

Prior to using the BioMark high throughput system, which can test as little material as a single cell against 96 genes, the group used conventional RT-PCR practices. The results from these techniques were limited as the methods used samples comprised of pooled cells or embryos, generating relative gene expression that represents an average of all cells assayed. Additionally using conventional techniques the Stanford researchers had not been able to discern between genes that are consistently differentially regulated versus those with a tendency towards stochastic changes or identify rare embryos. The reseachers concluded what they had accomplished on the BioMark system in 3 weeks would take more than 9 months to accomplish with conventional PCR methods.

A copy of the Research Spotlight 'Single-embryo gene expression for early embryo development' is now available free to download from http://www.fluidigm.com/pdf/fldm/FLDM_MRKT00117.pdf The Fluidigm BioMark System uses integrated fluidic circuit technology to reduce complexity, improve throughput with nanolitre precision and provide significant cost-savings (less reagent and smaller samples) for high-throughput PCR applications. Fluidigm's 48.48 Dynamic Array is capable of performing 2,304 simultaneous real-time PCR experiments running gold-standard TaqMan® assays in nanolitre quantities.

Fluidigm develops, manufactures and markets proprietary Integrated Fluidic Circuit (IFC) systems that significantly improve productivity in life science research. Fluidigm's IFCs enable the simultaneous performance of thousands of sophisticated biochemical measurements in extremely minute volumes. These "integrated circuits for biology" are made possible by miniaturising and integrating liquid handling components on a single microfabricated device. Fluidigm's IFC systems, consisting of instrumentation, software and single-use IFCs, increase throughput, decrease costs and enhance sensitivity compared to conventional laboratory systems. Fluidigm products have not been cleared or approved by the Food and Drug Administration for use as a diagnostic and are only available for research use.