Development of Laser Feedback Microscopy (LFM): High- Resolution Optical Imaging for Biological Science and Technology

激光反馈显微镜(LFM)是本实验室发展起来的一种新型扫描共聚焦光学显微镜。目前，我们已经生产了一种LFM，它可以测量z分辨率为纳米(10-9米)的低反射样品的表面形貌。LFM是一种远场光学显微镜技术，允许在生理条件下检查生物样品；不需要转移到高真空环境或施加其他特殊制备条件，例如染色或阴影。我们正在申请资金，以进一步开发和扩大原型仪器的建造，并将其应用于生物和其他材料的线性调频成像。该方案提出的主要目标是实现10-50 nm范围内的线性调频x，y分辨率，可与电子显微镜相媲美，并接近扫描隧道和原子力显微镜等表面探针技术的分辨率。此外，LFM具有用显微镜测量材料光学性质的能力；我们将利用这一独特的功能提供新的“光学对比”方法，从而使非传统的显微镜图像传达有关生物制剂的重要信息。最后，将建造一个具有尽可能高分辨率的全仪器、计算机控制台控制的线性调频装置，用于生物科学的普遍应用。

Laser feedback microscopy (LFM), a new type of scanning confocal optical microscopy, originated in this laboratory. Presently, we have produced a LFM which measures surface topography of low-reflectance samples with a z-resolution down to a nanometer (10-9 meter). LFM, a far-field optical microscopic technique, allows the examination of biological samples under physiological conditions; there is no requirement for transfer to a high-vacuum environment or the imposition of other special preparative conditions, e.g., staining or shadowing. We are requesting funds to further develop and extend the construction of prototype instruments and their application to LFM imaging of biological and other materials. The major goal presented in this proposal is the attainment of LFM x,y-resolution in the 10-50nm range, rivaling that of electron microscopy and approaching the resolution of surface-probe techniques such as scanning- tunneling and atomic-force microscopy. In addition, LFM has the capability of measuring microscopically the optical properties of materials; we will utilize this unique feature to provide new methods of "optical contrast" resulting in unconventional microscope images conveying important information about biological preparations. Finally, a fully- instrumented, computer-console controlled LFM with the highest possible resolution will be built for general use in the biological sciences.