Fluorescence microscopy with isotropic resolution using three objectives

Thomas Huelsnitz; Peter Kner

doi:10.1117/12.2213342

9 March 2016 Fluorescence microscopy with isotropic resolution using three objectives

Thomas Huelsnitz, Peter Kner

Proceedings Volume 9713, Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XXIII; 97130A (2016) https://doi.org/10.1117/12.2213342
Event: SPIE BiOS, 2016, San Francisco, California, United States

Abstract

Widefield and confocal fluorescence microscopy using a single objective suffer from poor resolution and a strong anisotropy between the lateral and axial resolution. Coherently combining the excitation and emission from two coaxial objectives improves the axial resolution up to sevenfold, but leaves the lateral resolution unchanged. Here we investigate the coherent combination of three objectives to create a point spread function (PSF) that is isotropic with higher resolution in the plane of the objectives. We develop a theoretical framework for simulating the performance of interferometric imaging with three objectives. Using three identical objectives with a large working distance and 0.9 numerical aperture (NA), the full-width half maximum of the confocal PSF is 135 nm compared to the lateral FWHM of 274 nm for imaging with a single objective at a wavelength of 515 nm.

Conference Presentation

Citation Download Citation

Thomas Huelsnitz and Peter Kner "Fluorescence microscopy with isotropic resolution using three objectives", Proc. SPIE 9713, Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XXIII, 97130A (9 March 2016); https://doi.org/10.1117/12.2213342

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