Standing Wave Total Internal Reflection Microscopy -- In Vivo Biological Imaging at the Nanometer Scale

该奖项将资助驻波全内反射显微镜(SW-TIRM)的开发。这种显微镜有可能提供比激发波长的十二分之一更好的横向分辨率。这项技术是基于在全内反射几何结构中形成高频驻波激励。使用一种简单的算法，可以从驻波不同相位的一系列图像重建高分辨率图像。特别有希望的是，能够以与扫描探针显微镜相当或更好的分辨率对生物标本进行非接触成像。为了在一维空间中演示这种技术的原理，我们已经建立了一个初步的SW-TIRM。在该奖项的资助下，这台当前仪器的能力将从一维扩展到2D和3D，使用散射光而不是荧光，并将数字反卷积和双光子方法融入图像分析。两名研究生将直接参与该仪器的开发。

This award will fund the development of a Standing Wave Total Internal Reflection Microscopy (SW-TIRM). This microscope has the potential to provide lateral resolution better than one twelfth of the excitation wavelength. This technique is based on the formation of high frequency standing wave excitation in a total internal reflection geometry. Using a simple algorithm, high-resolution images can be re-constructed from a series of images at different phases of the standing wave. Of particular promise is the ability to perform non-contact imaging of biological specimens at a resolution that is comparable to or better than that of scanning probe microscopy. A preliminary SW-TIRM has been constructed to demonstrate the principle of this technique in 1D. With funding from this award, the capabilities of this current instrument will be extended from 1D to 2D and 3D, use scattered light instead of fluorescence, and incorporate digital deconvolution and two photon methods to image analysis. Two graduate students will be directly involved in the development of this instrument.