SGER: New Techniques in Evanescent Wave Microscopy for Biological Applications

利用光激发和生物材料在固体表面操作的新方法，将开发新的技术来显着提高生物样品成像的灵敏度和分辨率。光激发在逝去波荧光中的可控穿透深度将与固体表面生物分子的有序阵列相结合，从而实现对特定于生物分子的分析物的高分辨率光学切片。通过将全内反射荧光(TIRF)与灵敏的垂直切片相结合，将克服两个重要障碍：首先，通过区分特定结合在表面的分析物和非特定结合的分析物，在固相分析中有望提高灵敏度一个数量级，从而提高选择性。其次，随着垂直切片的消逝荧光技术的扩展到前所未有的分辨率水平(大约10 nm)，将允许许多新的和强大的诊断技术的发展。利用现代光电子学和单模光波导将降低检测设备的复杂性和最终成本，使这些技术应用于各种生物分析分析成为可能。

New techniques to dramatically extend the sensitivity and resolution in imaging biological specimens using novel methods of optical excitation and manipulation of biological materials on solid surface will be developed. The controlled penetration depth of optical excitation in evanescent wave fluorescence will be combined with ordered arrays of biological molecules on solid surfaces, allowing high-resolution optical sectioning of analytes bound specifically to biomolecules. By so extending Total Internal Reflection Fluorescence (TIRF) with sensitive vertical sectioning, two significant hurdles will be overcome: First, by distinguishing between analytes bound specifically to the surface and those adhered non-specifically, enhancement in sensitivity by an order of magnitude and hence selectivity is expected in solid-phase assays. Second, the extension of evanescent fluorescence techniques with vertical sectioning to unprecedented resolution levels (in the order of 10 nm) will allow for the development of many novel and powerful diagnostic techniques. Utilizing modern optoelectronics and single-mode optical waveguides, the complexity and ultimate cost of the detection apparatus will be reduced, making possible the application of these techniques to a wide variety of bioanalytical assays.