Observation of biological objects using an optical diffraction tomographic microscope

Vincent Lauer

doi:10.1117/12.410638

18 December 2000 Observation of biological objects using an optical diffraction tomographic microscope

Vincent Lauer

Proceedings Volume 4164, Laser Microscopy; (2000) https://doi.org/10.1117/12.410638
Event: EOS/SPIE European Biomedical Optics Week, 2000, Amsterdam, Netherlands

Abstract

A prototype of a tomographic microscope has bene realized, which uses phase-shifting holography to sense the wave diffracted by an object. The observed object is successively illuminated by a series of about 1000 plane waves. Each diffracted wave yields a part of the frequency representation of the object, and superposition of these parts yields a 3D frequency representation. An inverse Fourier transform yields the 3D spatial representation, from which sections, projections or stereoscopic images can be extracted. This prototype has been used to image a variety of biological samples. Images show a resolution limit of about a quarter of a wavelength and a depth of field of about 40 microns. Important factors for the understanding of image characteristics include horizontal and vertical resolution, ability to image horizontal and/or vertical surfaces, ability to distinguish variations of refractive index from variations of absorptivity. These and other imaging characteristics of the tomographic microscope are discussed on the basis of a set of images of biological samples. The connection between characteristics of a three-dimensional image and its frequency representation is explained. Influence of an object's characteristics (thickness, refractive index, ...) on image quality is described. Possible improvements and their impact on image quality is also discussed.

Citation Download Citation

Vincent Lauer "Observation of biological objects using an optical diffraction tomographic microscope", Proc. SPIE 4164, Laser Microscopy, (18 December 2000); https://doi.org/10.1117/12.410638

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