| dc.description.abstract | Single molecule microscopy enables the direct observation of individual molecular events and is, therefore, an important tool in understanding the molecular mechanisms of disease pathogenesis. Analyzing single molecule data in order to extract biologically relevant information involves several complex data-processing steps, such as the identification of single molecules from microscopy images, estimating the location of identified molecules with high accuracy, determining the trajectory of individual molecules across time, etc. Such data-processing tasks are often mathematically complex, require specialized software, and are often not within the expertise of medical researchers and microscopists. Additionally, for accurate and biologically meaningful results, different approaches for each aspect of the data analysis need to be considered rather than a fixed sequence of data-processing steps.
Designing software tools to support the analysis of single molecule data is challenging. Diverse experiment types, complex data configurations, and a variety of processing tasks all need to be supported within analysis software while accommodating microscopists with varying levels of software expertise. Currently available analysis tools are often limited in these areas necessitating the use of several different tools in order to complete a single analysis.
Here, we present a software framework that addresses several of these challenges in the design of software tools for single molecule data analysis. We designed data structures using concepts of memory pointer-referencing and pointer-arithmetic to efficiently manage the large complex image data obtained from single molecule microscopy experiments and any associated data items, such as data-processing parameters, analysis results, etc. We developed a visual programming environment backed by a data-processing pipeline architecture to support flexibility in formulating analysis procedures for diverse data analysis scenarios. This environment allows users of all levels of software expertise to create and modify complex data-processing workflows. We developed a comprehensive framework for modeling and fitting single molecule images which is a critical element of estimating the location of single molecules with high accuracy. Important results obtained through studies conducted using this software framework regarding algorithms employed in the analysis of single molecule data are also presented. | en |
