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Ge J, Prasongtanakij S, Wood DK, et al. CometChip: a high-throughput 96-well platform for measuring DNA damage in microarrayed human cells. J Vis Exp. 2014;(92):e50607doi: 10.3791/50607.
Ge, J., Prasongtanakij, S., Wood, D. K., Weingeist, D. M., Fessler, J., Navasummrit, P., Ruchirawat, M., & Engelward, B. P. (2014). CometChip: a high-throughput 96-well platform for measuring DNA damage in microarrayed human cells. Journal of visualized experiments : JoVE, (92), e50607. https://doi.org/10.3791/50607
Ge, Jing, et al. "CometChip: a high-throughput 96-well platform for measuring DNA damage in microarrayed human cells." Journal of visualized experiments : JoVE vol. ,92 (2014): e50607. doi: https://doi.org/10.3791/50607
Ge J, Prasongtanakij S, Wood DK, Weingeist DM, Fessler J, Navasummrit P, Ruchirawat M, Engelward BP. CometChip: a high-throughput 96-well platform for measuring DNA damage in microarrayed human cells. J Vis Exp. 2014 Oct 18;(92):e50607. doi: 10.3791/50607. PMID: 25350601; PMCID: PMC4407627.
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J Vis Exp. 2014 Oct 18;(92):e50607. doi: 10.3791/50607.

CometChip: a high-throughput 96-well platform for measuring DNA damage in microarrayed human cells.

Journal of visualized experiments : JoVE

Jing Ge, Somsak Prasongtanakij, David K Wood, David M Weingeist, Jessica Fessler, Panida Navasummrit, Mathuros Ruchirawat, Bevin P Engelward

Affiliations

  1. Department of Biological Engineering, Massachusetts Institute of Technology; [email protected].
  2. Environmental Toxicology, Chulabhorn Graduate Institute.
  3. Department of Biomedical Engineering, University of Minnesota.
  4. Department of Biological Engineering, Massachusetts Institute of Technology.

PMID: 25350601 PMCID: PMC4407627 DOI: 10.3791/50607

Abstract

DNA damaging agents can promote aging, disease and cancer and they are ubiquitous in the environment and produced within human cells as normal cellular metabolites. Ironically, at high doses DNA damaging agents are also used to treat cancer. The ability to quantify DNA damage responses is thus critical in the public health, pharmaceutical and clinical domains. Here, we describe a novel platform that exploits microfabrication techniques to pattern cells in a fixed microarray. The 'CometChip' is based upon the well-established single cell gel electrophoresis assay (a.k.a. the comet assay), which estimates the level of DNA damage by evaluating the extent of DNA migration through a matrix in an electrical field. The type of damage measured by this assay includes abasic sites, crosslinks, and strand breaks. Instead of being randomly dispersed in agarose in the traditional assay, cells are captured into an agarose microwell array by gravity. The platform also expands from the size of a standard microscope slide to a 96-well format, enabling parallel processing. Here we describe the protocols of using the chip to evaluate DNA damage caused by known genotoxic agents and the cellular repair response followed after exposure. Through the integration of biological and engineering principles, this method potentiates robust and sensitive measurements of DNA damage in human cells and provides the necessary throughput for genotoxicity testing, drug development, epidemiological studies and clinical assays.

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