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

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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.