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Journal of Biomedical Optics

Dual-modal three-dimensional imaging of single cells with isometric high resolution using an optical projection tomography microscope
Author(s): Qin Miao; J. Richard Rahn; Anna V. Tourovskaia; Michael G. Meyer; Thomas Neumann; Alan C. Nelson; Eric J. Seibel
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Paper Abstract

The practice of clinical cytology relies on bright-field microscopy using absorption dyes like hematoxylin and eosin in the transmission mode, while the practice of research microscopy relies on fluorescence microscopy in the epi-illumination mode. The optical projection tomography microscope is an optical microscope that can generate 3-D images of single cells with isometric high resolution both in absorption and fluorescence mode. Although the depth of field of the microscope objective is in the submicron range, it can be extended by scanning the objective's focal plane. The extended depth of field image is similar to a projection in a conventional x-ray computed tomography. Cells suspended in optical gel flow through a custom-designed microcapillary. Multiple pseudoprojection images are taken by rotating the microcapillary. After these pseudoprojection images are further aligned, computed tomography methods are applied to create 3-D reconstruction. 3-D reconstructed images of single cells are shown in both absorption and fluorescence mode. Fluorescence spatial resolution is measured at 0.35 μm in both axial and lateral dimensions. Since fluorescence and absorption images are taken in two different rotations, mechanical error may cause misalignment of 3-D images. This mechanical error is estimated to be within the resolution of the system.

Paper Details

Date Published: 1 November 2009
PDF: 6 pages
J. Biomed. Opt. 14(6) 064035 doi: 10.1117/1.3275470
Published in: Journal of Biomedical Optics Volume 14, Issue 6
Show Author Affiliations
Qin Miao, Univ. of Washington (United States)
J. Richard Rahn, VisionGate, Inc. (United States)
Anna V. Tourovskaia, VisionGate, Inc. (United States)
Michael G. Meyer, VisionGate, Inc. (United States)
Thomas Neumann, VisionGate, Inc. (United States)
Alan C. Nelson, VisionGate, Inc. (United States)
Eric J. Seibel, Univ. of Washington (United States)

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