Grant: Research Tools and Instruments

To understand the mechanisms underlying fundamental biological processes such as microbial replication and cell differentiation, it is essential to be able to efficiently and reliably analyze samples of proteins and nucleic acids, both qualitatively and quantitatively. For example, to follow cell signaling events it is often necessary to quantify posttranslational modifications such as phosphorylation and ubiquitination. Immunoblotting is a standard approach for studying such modifications; however, to obtain high quality data, gel documentation systems are required for efficient acquisition of quantitative data that can be subjected to statistical analyses. Moreover, it is often necessary to assess both the global levels of a particular protein and levels of a specific modified form, as well as levels of loading controls. In addition to protein detection, the imaging and analysis of nucleic acids that are resolved using agarose or acrylamide gel electrophoresis form the backbone of molecular biology. This proposal is to acquire a bioimaging system for proteins and nucleic acids to support fundamental research programs in the biological sciences in a pavilion for 12 laboratories in microbiology and immunology research. The requested imaging module for proteins is a state-of-the art laser scanner module with the capacity for multiplex fluorescence imaging and chemiluminescence imaging. It will be complemented with a work-horse module to support routine nucleic acid imaging and data analysis using standard fluorescent stains. The system will be operated from a computer with proprietary software for image acquisition and analysis. Multiple software licenses will be included to ensure trainees can analyze data at their respective workstation, and thus use of the imaging system will be reserved for data acquisition. There is an urgent need for this system as the nucleic acid imager in the pavilion is past its useful life, due to outdated software and computer operating systems that cannot be updated. Moreover, for immunoblot experiments researchers in the pavilion are presently relying on a decades old film developer for which it has been impossible to order replacement parts for several years. The acquisition of a biomolecular scanner that allows researchers to conduct multiplex immunoblotting will provide higher quality data than is possible with the present infrastructure in the pavilion. It will also ensure that trainees acquire research experience using state-of-the-art approaches to quantitatively analyze specific proteins and nucleic acids in the context of their experiments, while incorporating best practices for the inclusive presentation of data in visual format, so that as many individuals as possible with vision differences can access the data.