This NSF award, made to the Brain Networks Laboratory at Texas A&M University, will help enhance and transform the Knife-Edge Scanning Microscope (KESM), a "one of a kind" microscopy instrument, into a more widely available system for biological research. KESM is capable of sectioning and imaging whole small animal organs at submicrometer resolution (that is, in very fine detail, smaller than the size of a single cell). A prototype KESM instrument was constructed, and its capability was successfully demonstrated by scanning diverse biological organs including whole mouse brains, octopus brains, and the mouse lung at submicrometer resolution. This project will enhance this prototype significantly, and transform it into a robust system that can be replicated and operated with ease by other research groups and industry partners. The enhanced instrument to be built by this project addresses at least two major emerging directions in biological research: (1) "omics" (the study of a total collection of some biological entity, for example, genomics for genes) and (2) multi-scale modeling (modeling systems across a range of scales, for example from the cell up to the whole organ). With the tremendous success of genomics in the 1990's and beyond, biological research is increasingly moving toward various forms of omics. Much of the omics research depends on anatomical information (e.g., connectomics, studying the complete wiring diagram in the brain) or genomic information (e.g., gene expression levels) at the whole-organ-level. Multi-scale modeling has also become a major methodological process in biological research. In many projects in omics and multi-scale modeling, sub-micrometer microscopy data from whole biological organs are essential yet available tools are unable to meet the current demand. KESM is expected to fill this gap. The specific enhancements of the instrument include: (1) enhanced imaging (higher resolution optics and camera, fluorescence imaging [key to genetic and molecular studies]), (2) enhanced mechanical control (more rigid, accurate, and higher resolution motion control), (3) enhanced cutting (vibrating knife, real time monitoring), and (4) enhanced robustness (quality monitoring, calibration). The overall resulting improvement can be summarized as follows: (1) 3X improvement in imaging resolution, (2) 10X improvement in robustness of operation, (3) 10X improvement in imaging speed (compared to competing methods), (4) new fluorescence imaging capability.
Broader impacts: (1) Impact on the research community: The enhanced KESM will allow researchers to obtain high resolution, whole-organ data for multi-scale, omics investigation of various types of biological organs. (2) Impact on education: Microscopic atlases of whole biological organs, such as the web-based KESM mouse brain atlas developed by the project team, will serve as an educational resource for students and educators at all levels (K-12, undergraduate, graduate, and general public). As part of this project, graduate and undergraduate students will be trained in a multidisciplinary environment (neuroscience and computer science). (3) Instrument dissemination plan: The project team will collaborate with a start-up company, to streamline system integration and manufacturing of the KESM instrument for broader dissemination. The design of the instrument and the operational instructions will be made available to the biological research community for those who wish to build their own instrument.
