SGER: New Techniques in Evanescent Wave Microscopy for Biological Applications

SGER：倏逝波显微镜的生物应用新技术

• 批准号: 9812377
• 负责人: Bennett Goldberg
• 金额: $ 11万
• 依托单位: Trustees of Boston University
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 1998
• 资助国家: 美国
• 起止时间: 1998-06-01 至 2001-05-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=9812377&HistoricalAwards=false
• 关键词: SGER; New; Techniques; Evanescent; Wave

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.

将开发新的技术，以显着提高灵敏度和分辨率，在成像生物标本使用新的方法的光激发和操纵的生物材料在固体表面上。倏逝波荧光中光激发的受控穿透深度将与固体表面上生物分子的有序阵列相结合，从而允许对与生物分子特异性结合的分析物进行高分辨率光学切片。通过这样扩展全内反射荧光（TIRF）与灵敏的垂直切片，两个显著的障碍将被克服：首先，通过区分特异性结合到表面的分析物和那些非特异性粘附的分析物，在固相测定中预期灵敏度提高一个数量级，因此选择性提高。第二，将具有垂直切片的渐逝荧光技术扩展到前所未有的分辨率水平（在10 nm的量级）将允许开发许多新的和强大的诊断技术。利用现代光电子学和单模光波导，检测装置的复杂性和最终成本将降低，从而使这些技术应用于各种生物分析测定成为可能。