Streamlining Microscopy Assay Development: A Comprehensive Workflow Solution for Enhanced Reproducibility and Efficiency

Project Summary Microscopy users have widely varying levels of expertise and operate in the design and execution of high complexity assays. Microscopy assay design can be highly challenging due to several key factors including a growing number of reagents to select from (including functional dyes, fluorophores and primary/secondary antibodies), significant variety in instrument components and configurations, and increased experimental complexity with increasing numbers of markers/labels/dyes being included in each experiment (a movement to multiplex experiments). For many researchers, these requirements can be a barrier to entry and an incomplete understanding or poor execution can result in improper assay design, poor experimental outcomes, wasted resources (including rare or low-volume resources), and poor reproducibility. Novel approaches that support researchers of all skill levels (including early stage through expert scientists) to design better experiments and select instrument components and reagents that work together has significant potential to reduce costs associated with wasted resources, improve experimental outcomes, and potentially address one aspect of the reproducibility crisis. To address this unmet need, FluoroFinder LLC is advancing a novel microscopy workflow tool to streamline assay development, improve awareness and education around optimal selection of assay components (instruments and reagents), and improve reproducibility in the field. The FluoroFinder Microscopy tool is a software-based solution to guide users through a structured workflow that addresses the most common challenges in microscopy including optimizing microscope component configuration and selecting validated reagents that are appropriate for a specific instrumental configuration and biological experiment. This will enable users to identify optimal antibodies, fluorophores, functional dyes, and ancillary reagents for their biological experiment and instrument set-up. Prior to this Phase I proposal, FluoroFinder has built an extensive database of fluorescent reagents and antibodies that includes more than 3 million entities aggregated from vendor databases that are relevant to microscopy applications. Building on the success of a previously developed industry-leading automated flow cytometry panel builder tool (IntelliPanel), FluoroFinder has initiated development of a user interface (UI)/user experience (UX) that enables matching of fluorophores to primary/secondary antibodies for use in microscopy. The goal of this Phase I project is to develop and demonstrate feasibility for the FluoroFinder Microscopy Tool, where FluoroFinder will complete prototype development of the UI/UX that supports microscopy experimental design with a guided process that includes instrument configuration settings and reagent selection. The UI/UX will include support for epi-fluorescent and confocal microscopy design, as well as cyclic immunofluorescence (CycIF) that supports the latest spatial biology instruments and methods. Commercialization of the Microscopy Tool would significantly impact research and medical diagnostics, enabling researchers of all skill levels to identify optimal reagents for their experiments.