Semi-Automated Analysis Software for a Novel Biochemistry Assay
AuthorVesco, Joseph Michael
AdvisorHarris, Frederick C
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Some of the work done in the Baker lab at the University of Nevada, Reno hasbeen directed towards the analysis of muscle tissue in a single molecule arrangement.The process is a novel high-throughput single molecule binding assay, or SiMBA forshort. This assay is performed by binding the myosin molecules of the muscle tissueto a coverslip and looking at how the fluorescently labeled actin filaments bind tothese myosin as imaged with a fluorescence microscope. The conditions are variedand the effects on actin-myosin binding kinetics are observed. In order to analyzethe binding times and unbound times a researcher must observe the interactions andmanually collect the data. This particular method of data collection is tedious andtime consuming thereby making this portion of the experiment the "rate limiting"factor for producing results in a timely manner. As this can take many hours toanalyze a one minute long movie an automated or semi-automated solution wouldbe beneficial to this assay. This thesis presents the design and implementation ofa semi-automated solution for identifying and tracking a variable number of objectsthat exhibit a multitude of behavior, and extracting the specific behaviors of motionand stagnation as well as the duration of these behaviors.