Recipient Organization
RUTGERS, THE STATE UNIVERSITY OF NEW JERSEY
3 RUTGERS PLZA
NEW BRUNSWICK,NJ 08901-8559
Performing Department
Food Science
Non Technical Summary
Dissecting the molecular mechanisms that support biological processes important for food and nutrition, for agriculture, and for the environment involves a complex interplay of many proceses: the identification of the molecular actors (proteins, lipid membranes, nucleic acids, small molecules) involved in the mechanism; the generation of specific physical models describing how these actors behave and interact to generate a specific biological mechanism; and the testing of these models for their correspondence to molecular reality. The final testing phase often presents significant difficulities associated with identifying the precise roles engaged in by specific actors. Does protein A initiate a process or follow the action of another molecule? Does the nucleic acid interact with protein B or another protein? Do the proteins bind to the membrane while functioning? Etc., etc.The fluorescence microscope purchased in this project will facilitate such precise testing of physical mechanisms by allowing researchers to actually monitor the behaviour of single individual molecular actors engaged in a specific biological function, either in a model system in vitro or even perhaps in a living cell. This instrument has the capability to detect, locate, and monitor the movement of single molecules that either include an intrinsice fluorescence probe or that can be covalently labeled with a fluorescence probe. Such molecular precision thus allows researchers to precisely test the role played by specific molecules, be they lipids, proteins, nucleic acids or whatever, in the specific mechanisms of biological processes.This project will thus make simgle molecule imaging readily available to a novel research audience in food & nutrition, agriculture, and the environment. These capabilities should have a dramatic impact on the state of knowledge in these fields and inspire additional studies of biological mechanisms molecule-by-molecule.
Animal Health Component
0%
Research Effort Categories
Basic
90%
Applied
0%
Developmental
10%
Goals / Objectives
Advances in instrumentation and technique have pushed the spatial resolution of optical microscopy below the physical diffraction limit of approximately half the optical wavelength (~250 nm); when combined with sensitive detectors, it is now routine with proper instruments to monitor and spatially resolve individual fluorescent molecules with resolutions of 10s of nanometers. Extensive control of single, often site-specific labeling of proteins, nucleic acids, and other biomolecules combined with single molecule imaging now enables precision monitoring of the spatial location, physical structure and behavior, and biological interactions of targeted single molecules on surfaces, in solution, and within live cells. This project will acquire and implement the use of a fluorescence microscope that enables single molecule detection on the George H Cook Campus of Rutgers School of Environmental and Biological Sciences (SEBS) in New Brunswick. This instrument will enable a variety of single molecule measurements by approximately 16 researchers in six different departments at the School and by a number of other users from other Schools across Rutgers University--New Brunswick.The goal of this one year project is thus the acqusition and installation of a Nanoimager high resolution fluorescence microscope and the education and training of a core group of users on this instrument whose expertise is in the fields of food and nutrition, agriculture, and environmental science. The objectives of each distinct project that uses the Nanoimager is specific to each individual researcher.
Project Methods
This project will make use of a variety of methods that optimize the fluorescence-based dectection of, localization of, structure of, as well as the quantification of the specific properties of single molecules. The experimental basis for the method is the labeling of individual molecules, whether sugars, carbohydrates, proteins, lipids, or nucleic acids, with robust and appropriate fluorescence labels (or probes). The sensitivity of the instrument allows these labeled molecules to be detected one-by-one at surfaces, in solutions, or even in living cells when present at very low concentrations (<< nM.) The data collected include number and location of molecules, trajectories of their movements, possible interactions with other molecules, etc. The use of two or more differently labled molecules allows one to monitore specific molecular interactions at the single molecule level and various methods for doubly labeling individual molecules provides insight into molecular structure and changes in molecular structure that accompany specific processes. The specific advantage of the instrument is thus that it enables researchers to analyze and dissect molecule-by-molecule the molecular bases for biologcal functions. Such specificity of information enables the precision testing of physical details of models for biological mechanisms at the highest resolution possible, at that of the behavior of individual molecules.