Developing a High-Throughput In Vitro DNA Damage Assay

The Engelward Lab in collaboration with Sangeeta Bhatia's Lab have developed a high throughput DNA damage analysis platform. Each well of a 96-well plate contains patterned microwells for single cell capture and DNA damage quantification. The new technique is based on the 30-year-old "comet assay" --named for the comet-shaped morphology that damaged DNA forms when pulled during electrophoresis. The new technology will enable the team to conduct both large scale epidemiological and clinical studies.
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(1) Patterning Cells Using Microfabrication Techniques -- Using the microfabrication expertise of the Bhatia Lab, we developed a method to pattern microwells in agarose for capture of single cells in a patterned array.

(2) Software Development for High Throughput Analysis -- We developed software to automatically image and analyze our comet assay platform. The software takes advantage of the arrayed cells in order to unbiasedly eliminate contamination. In addition, the software takes advantage of the self-calibrating property of the microwells for accurate determination of the tail-to-head threshold.
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(3) High Throughput DNA Damage Analysis -- The Engelward Lab is applying our new technology in a variety of clinical and epidemiological settings. We are working with researchers in Thailand to study the effects of air pollution on school children in high risk regions, compared to inhabitants of cleaner areas. We are also collaborating with researchers at Boston University Medical School to investigate the consequences of cigarette smoking. Ultimately, we aim to provide an assay to the greater research community that is simple enough to be useful in a broad range of clinical, epidemiological, and experimental studies.