An automated dielectrophoretic-based single cell separation technique to improve laboratory efficiency, mixture deconvolution and combat sample inhibition

As submitted by the proposer: This 12-month effort will address several core needs in the field of forensic DNA analysis, (1) the development of a new, reliable and robust method for the identification of spermatozoa, (2) development of a simple, efficient collection method for sperm and epithelial cell fractions impacting downstream analysis and (3) the ability to preferentially collect and concentrate low template samples without the need for DNA quantitation. Within forensic biology/DNA analysis, much effort has focused on the processing of DNA evidence following serological analysis as opposed to improvements in serological techniques that may positively impact efficiency and enhance downstream DNA analysis. Current methodologies, such as light microscopy, laser capture microdissection and subsequent differential extraction, are generally accepted by the scientific community and considered highly reliable. However, they are laborious and are many times fail to adequately separate human spermatozoa from epithelial cells, leading to the need for DNA mixture interpretation (Miller et. al. 2011). In addition, there are continual needs to enhance the analysis techniques addressing low template number and inhibitor-containing samples. The DEPArray™ system (Silicon Biosystems) is a novel multipurpose instrument based on dielectrophoretic principles that has the ability to identify, isolate, and recover individual cells from heterogeneous samples (Silicon Biosystems 2013) The predominant use of the DEParray exists in cancer research to isolate and separate circulating tumor cells (CTCs) from white blood cells (WBCs). We propose that the DEParray may also be used in the forensic biological assays to specifically address the aforementioned core areas of need. This project contains a research and/or development component, as defined in applicable law. ca/ncf