Research Project
10175807
PROJECT SUMMARY This proposal seeks funding for a Vectra® Polaris? Automated Quantitative Imaging System (Polaris?), a state- of-the-art multispectral imaging microscope to serve multiple NIH-supported projects within the Mycobacteria Research Laboratories (MRL) at Colorado State University (CSU). Our collective mission is to develop effective chemotherapies and vaccines for mycobacterial diseases affecting millions worldwide. The biological activity and safety profile of candidate vaccines and treatments can only be fully understood through characterizing the host immune response, primarily by evaluating large networks of immune biomarkers through flow cytometry and immunohistochemistry. MRL researchers are seeking to visualize the spatial expression of immune biomarkers by multiplex staining of intact tissue samples obtained from preclinical models of mycobacterial diseases. Until now, microscopic analysis of biomarkers through confocal and traditional immunofluorescence was drastically limited by autofluorescence in tissue (especially in formalin fixed and diseased tissue) and low signal-to-noise ratios. The Polaris? system has a unique set of microscopic capabilities that combined with the inForm® workstation allows simultaneous identification and quantification of multiple targets and their spatial relationships in tissue sections and microarrays. A critical aspect of the PolarisTM multispectral technology is the ability to readily distinguish autofluorescence from positive biomarker signal, improving the signal-to-noise ratio substantially. This allows for a more sensitive and accurate analysis of samples and enables true cell-to-cell interactions to be revealed. The system is user-friendly, and its automated scanning system allows for high speed digital and whole-slide scanning between 10-40x in brightfield or fluorescence. The nature of the illumination system significantly decreases deleterious effects such as photobleaching and photodamage, and thus specimens can be imaged for longer periods of time than with, for example, a confocal microscope. Furthermore, the advanced machine-learning based approaches of the included software automatically segment and quantitate tissue structures. The Polaris? system will be incorporated into our existing microscopy core facility in order to expand access to trainees and other research groups at CSU and in Northern Colorado, while prioritizing access for the major users. Incorporation into the foundational core will sustain long term operation and provide consistent oversight and training. In summary, this state-of-the-art imaging technology is of critical importance to development of chemotherapies and vaccines by the MRL and will add value to multiple other NIH-supported projects at CSU.
