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Comprehensive Profiling of miRNAs in Stem Cells Using Microfluidics.
Fluidigm Europe has announced the availability of a new customer technical poster** entitled 'Comprehensive profiling of microRNAs in murine hematopoietic stem cells using a microfluidics approach' that reports upon the pioneering work of researchers at the BC Cancer Agency and University of British Columbia (Vancouver, Canada).
MicroRNAs (miRNAs) have been shown to be key regulators of hematopoiesis. However, the detailed profile of miRNA expression throughout hematopoietic development has not been charted, owing primarily to challenges in obtaining highly purified samples of stem and progenitor cells as well as profiling hundreds of miRNAs from low cell numbers. To overcome these limitations, the new technical poster describes how microfluidic technologies were combined with a 288-plex real time PCR approach, allowing the detection of 288 miRNAs in small numbers (c. 3000) of cells.
The poster describes how over twenty unique murine hematopoietic cell populations were isolated through FACS sorting, including hematopoietic stem cells (HSCs), myeloid and lymphoid progenitor cells and mature lineage populations. Several thousand nanolitre volume quantitative PCR reactions were performed using Fluidigm BioMark™ 48.48 Dynamic Arrays and miRNA specific TaqMan® probes. For each miRNA, synthetic miRNA standards were used to determine the absolute number of miRNA molecules per cell. 133 of the 288 miRNAs tested were detected across all cell populations.
The researchers conclude that Fluidigm's dynamic array provides a rapid, sensitive and cost-effective method for miRNA profiling in rare cell populations. The sensitivity of Fluidigm's dynamic array is shown to detect heterogeneity in miRNA expression at the single cell level in purified populations.
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 miniaturizing and integrating liquid handling components on a single microfluidic device (IFC). Fluidigm's systems, consisting of instrumentation, software and single-use chips, 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.
For more information, please visit www.fluidigm.com
A copy of the technical poster is available on request by contacting biomarkeurope@fluidigm.com
** Nominated as best poster at RNAi Europe, September 2009.
* Fluidigm, the Fluidigm logo, Topaz, BioMark, Dynamic Array, Digital Array, Access Array and NanoFlex are trademarks or registered trademarks of Fluidigm Corporation. All other trademarks are the property of their respective owners.
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