The Swept Field Confocal Microscope

DESCRIPTION (provided by applicant): A primary goal in neuroscience is to understand at a mechanistic level the functioning of neuronal networks with specific behaviors. This involves in general, the anatomical connectivity among the various types of neurons within the local network, the physiological properties of these cells, and the spatiotemporal firing signals of the neurons within the circuit. The challenge is not only to discover the connectivity among neurons but also to effectively record the signal activity within the neuronal network during a behavior in such a way as to provide functional information. Since 2001 Prairie Technologies, Inc. has been designing, building, refining and selling a new type of confocal scanning system, the Swept Field Confocal microscope (SFC). SBIR Phase I and II awards have provided the funding necessary for Prairie to develop an entry level product for the biological marketplace. Due to successful completion of all of our Phase I and Phase II goals, we have been able to sell over 60 units totaling to sales of 6,000,000.00 for the company. Recently we have been able to develop a worldwide sales channel for the SFC with Nikon Instruments, Inc. Because of high speed and low toxicity, the SFC is now gaining in sales popularity in the neuro and cell biology fields. In order to provide more functional information, we propose to further improve the speed of the SFC and simplify its ease of use for the scientists and to increase the amount of applications that can be performed with the instrument such as Fluorescence Resonance Energy Transfer (FRET). In particular, we would like to add quantitative techniques such as Fluorescence Lifetime Imaging (FLIM) for the study of dynamic processes and FRET analysis. By completing the specific aims outlined in this proposal, the SFC can not only perform as a confocal microscope, but have capability to provide dynamic information about the local neuron network with high resolution. In addition, with completion of this project Prairie can achieve increased product sales of the SFC into the 200 million annual world-wide laser scanning microscopy market. PUBLIC HEALTH RELEVANCE: A primary goal in neuroscience is to understand at a mechanistic level the functioning of neuronal networks with specific behaviors, which can be used to characterize neurological disorders. Such studies require the ability to identify the signaling networks and noninvasively study the dynamics within living cells. The improved Swept Field Confocal microscope system will provide such capabilities to record the spatiotemporal information in real time of normal and diseased processes and to study the cellular dynamics with additional information about the microenvironment and signaling pathways detected with Fluorescent Lifetime Imaging.