Automation of Differential Extraction with Sperm Quantitation usingMicrofluidic-Integrated Shadow Imaging System for Forensic Applications

As submitted by the proposer: Rapid and efficient processing of sexual assault evidence will accelerate forensic investigation and decrease casework backlogs. Therefore, the standardized protocols currently used in forensic laboratories will need continued innovation to handle the increasing number and complexity of samples being submitted to forensic labs. To our knowledge, currently no rapid differential extraction technology, which includes both sperm isolation and quantitation is available. Here, we propose to develop a novel sperm capture and differential extraction microchip-integrated system with a novel holography imaging platform. The microchips will (i) selectively capture sperm from sexual assault samples, (ii) be sensitive within the forensic cut-off (with ±10% error range), provide cost effective solution and (iii) handle small volume of sample (<100 ìL). The holographic imaging platform will then rapidly provide the results within 10 minutes of image capture and analysis to aid downstream forensic analysis. To fulfill our goal, we will complete the following specific aims. We will first develop a portable lensless system for holographic imaging and microchips with a unique sperm capture reagent. To further develop an integrated reagent, hardware and software system with disposable microchips to capture and quantitate the number of sperm in a forensic sample and to develop specific protocols for forensic applications with the system (Aim 1). We will then evaluate the holographic imaging platform and microfluidic platform with a statistically relevant number of adjudicated sexual assault samples through our collaboration with the Broward Sheriff’s Office Forensic Laboratory (Aim 2). We will conduct a Developmental Validation of the integrated microfluidic imaging system including sensitivity, precision and accuracy, reproducibility, stability studies and correlation with currently used methods, resulting in commercialization of a next generation system to accelerate the differential extraction process for forensic investigations (Aim 3). This project contains a research and/or development component, as defined in applicable law. ca/ncf