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Nat Methods. 2012 Nov;9(11):1101-6. doi: 10.1038/nmeth.2185. Epub 2012 Sep 30.

A microfluidic device and computational platform for high-throughput live imaging of gene expression.

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  • 1Department of Biology, Duke University, Durham, North Carolina, USA.

Abstract

To fully describe gene expression dynamics requires the ability to quantitatively capture expression in individual cells over time. Automated systems for acquiring and analyzing real-time images are needed to obtain unbiased data across many samples and conditions. We developed a microfluidics device, the RootArray, in which 64 Arabidopsis thaliana seedlings can be grown and their roots imaged by confocal microscopy over several days without manual intervention. To achieve high throughput, we decoupled acquisition from analysis. In the acquisition phase, we obtain images at low resolution and segment to identify regions of interest. Coordinates are communicated to the microscope to record the regions of interest at high resolution. In the analysis phase, we reconstruct three-dimensional objects from stitched high-resolution images and extract quantitative measurements from a virtual medial section of the root. We tracked hundreds of roots to capture detailed expression patterns of 12 transgenic reporter lines under different conditions.

PMID:
23023597
[PubMed - indexed for MEDLINE]
PMCID:
PMC3492502
Free PMC Article

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