Extended resolution wide-field optical imaging: objective-launched standing-wave total internal reflection fluorescence microscopy |
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Authors: | Chung Euiheon Kim Daekeun So Peter T C |
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Institution: | Department of Mechanical Engineering, Massachusetts Institute of Technology, Cambridge 02139, USA. ogong50@mit.edu |
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Abstract: | Standing-wave total-internal-reflection fluorescence (SW-TIRF) microscopy uses a super-diffraction-limited standing evanescent wave to extract the high-spatial-frequency content of an object through a diffraction-limited optical imaging system. The effective point-spread function is better than a quarter of the emission wavelength. With a 1.45 numerical aperture objective and 532 nm excitation wavelength, a Rayleigh resolution of approximately 100 nm can be achieved, which is better than twice the resolution of conventional TIRF microscopy. This first experimental realization of SW-TIRF in an objective-launched geometry demonstrates the potential for extended resolution imaging at high speed by using wide-field microscopy. |
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