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

Publication ,  Journal Article
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 in: Nat Methods
November 2012

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.

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Published In

Nat Methods

DOI

EISSN

1548-7105

Publication Date

November 2012

Volume

9

Issue

11

Start / End Page

1101 / 1106

Location

United States

Related Subject Headings

  • Plant Roots
  • Microscopy, Confocal
  • Microfluidic Analytical Techniques
  • Gene Expression Regulation, Plant
  • Developmental Biology
  • Arabidopsis
  • 31 Biological sciences
  • 11 Medical and Health Sciences
  • 10 Technology
  • 06 Biological Sciences
 

Citation

APA
Chicago
ICMJE
MLA
NLM
Busch, W., Moore, B. T., Martsberger, B., Mace, D. L., Twigg, R. W., Jung, J., … Benfey, P. N. (2012). A microfluidic device and computational platform for high-throughput live imaging of gene expression. Nat Methods, 9(11), 1101–1106. https://doi.org/10.1038/nmeth.2185
Busch, Wolfgang, Brad T. Moore, Bradley Martsberger, Daniel L. Mace, Richard W. Twigg, Jee Jung, Iulian Pruteanu-Malinici, et al. “A microfluidic device and computational platform for high-throughput live imaging of gene expression.Nat Methods 9, no. 11 (November 2012): 1101–6. https://doi.org/10.1038/nmeth.2185.
Busch W, Moore BT, Martsberger B, Mace DL, Twigg RW, Jung J, et al. A microfluidic device and computational platform for high-throughput live imaging of gene expression. Nat Methods. 2012 Nov;9(11):1101–6.
Busch, Wolfgang, et al. “A microfluidic device and computational platform for high-throughput live imaging of gene expression.Nat Methods, vol. 9, no. 11, Nov. 2012, pp. 1101–06. Pubmed, doi:10.1038/nmeth.2185.
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. A microfluidic device and computational platform for high-throughput live imaging of gene expression. Nat Methods. 2012 Nov;9(11):1101–1106.

Published In

Nat Methods

DOI

EISSN

1548-7105

Publication Date

November 2012

Volume

9

Issue

11

Start / End Page

1101 / 1106

Location

United States

Related Subject Headings

  • Plant Roots
  • Microscopy, Confocal
  • Microfluidic Analytical Techniques
  • Gene Expression Regulation, Plant
  • Developmental Biology
  • Arabidopsis
  • 31 Biological sciences
  • 11 Medical and Health Sciences
  • 10 Technology
  • 06 Biological Sciences