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

Published

Journal Article

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.

Full Text

Duke Authors

Cited Authors

  • Busch, W; Moore, BT; Martsberger, B; Mace, DL; Twigg, RW; Jung, J; Pruteanu-Malinici, I; Kennedy, SJ; Fricke, GK; Clark, RL; Ohler, U; Benfey, PN

Published Date

  • November 2012

Published In

Volume / Issue

  • 9 / 11

Start / End Page

  • 1101 - 1106

PubMed ID

  • 23023597

Pubmed Central ID

  • 23023597

Electronic International Standard Serial Number (EISSN)

  • 1548-7105

Digital Object Identifier (DOI)

  • 10.1038/nmeth.2185

Language

  • eng

Conference Location

  • United States