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Automated Fluorescence Detection and Imaging of RNA Species in Live Cells
EP22396
Poster Title: Automated Fluorescence Detection and Imaging of RNA Species in Live Cells
Submitted on 08 Oct 2014
Author(s): Paul Held1, Victor Koong2, Don Weldon2, and Peter Banks1
Affiliations: 1BioTek Instruments, Inc. 2EMD Millipore
This poster was presented at MipTec 2014
Poster Views: 1,790
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Poster Information
Abstract: disease directed research relies on the ability to rapidly make assay determinations on large numbers of samples. At the same time phenotypic information is also desired to assess the true cellular response. Towards that end, having multiple fluorescent probes capable of simultaneously detecting different cellular RNAs in live cells are of particular importance.

Here we describe the use of a combination microplate reader and imager to detect changes in RNA levels using a series of gold nanoparticle fluorescent probes (SmartFlare™ Probes from EMD Millipore). The multi-mode microplate reader is capable of digital microscopy and conventional microplate detection. Probes consist of a gold nanoparticle conjugated with multiple copies of double stranded oligonucleotides. The longer strand is complementary to the RNA target where as the shorter reporter strand contains a fluorescent molecule (CY3 or CY5) that is quenched when in proximity to the gold core. With exposure to the target RNA the reporter strand is displaced as the target strand binds to its complimentary strand located on the probe. Displacement removes the proximity associated quenching resulting in fluorescence. The degree of fluorescence is dependent on the amount of target RNA present.
Summary: The determination of intracellular RNA levels is a critical component in elucidating the cellular responses of living cells to external stimuli. Many of the techniques traditionally used for RNA quantitation involve transfection, laborious sample preparation and RNA amplification, which can preclude large sample numbers. References: Report abuse »
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