Developments in three-dimensional stereo brightfield microscopy

We present recent developments of a widefield computer/microscope system and image reconstruction algorithm for producing three‐dimensional (3D) increased depth of field images in the form of brightfield stereo pairs of thick specimens. The theoretical principle of this image reconstruction techniqu...

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Veröffentlicht in:Microscopy research and technique 1993-04, Vol.24 (5), p.437-451
Hauptverfasser: Willis, Byron, Turner, James N., Collins, Doris N., Roysam, Badrinath, Holmes, Timothy J.
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container_end_page 451
container_issue 5
container_start_page 437
container_title Microscopy research and technique
container_volume 24
creator Willis, Byron
Turner, James N.
Collins, Doris N.
Roysam, Badrinath
Holmes, Timothy J.
description We present recent developments of a widefield computer/microscope system and image reconstruction algorithm for producing three‐dimensional (3D) increased depth of field images in the form of brightfield stereo pairs of thick specimens. The theoretical principle of this image reconstruction technique is based on Weiner‐type inverse filtering. A number of extensions and refinements to our previous work have included further testing of the system with a broader class of specimens and the implementation of several pragmatic refinements important for future 3D microscopy systems. These refinements include histogram modification routines for improving visualization, a preprocessing routine to eliminate edge artifacts due to circular convolution and other effects, stereo viewing angle optimization, a rule of thumb estimate for the axial sampling rate, and incorporation of a variation of the Fast Fourier Transform and filtering operations that significantly reduce computational time. Images of spyrogyra, neonatal rat hippocampal neurons, and cervical/vaginal cell smears are presented to show the utility of these methods for 3D visualization. The primary advantages of these methods are that they operate with an ordinary transmitted light microscope and are inexpensively implemented on a personal computer with reasonable computation time. © 1993 Wiley‐Liss, Inc.
doi_str_mv 10.1002/jemt.1070240509
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source MEDLINE; Wiley Online Library Journals Frontfile Complete
subjects Algorithms
Animals
Cervix Uteri - cytology
Computer/microscope system
Conventional optical microscopes
Depth of field
Eukaryota - cytology
Exact sciences and technology
Fast Fourier Transform
Female
Fourier Analysis
Hippocampus - cytology
Humans
Image Processing, Computer-Assisted
Image reconstruction
Instruments, apparatus, components and techniques common to several branches of physics and astronomy
Inverse filtering
Microscopy - instrumentation
Microscopy - methods
Missing cone
Neurons - cytology
Optical instruments, equipment and techniques
Optical sectioning
Optical transfer function
Physics
Stereo pairs
title Developments in three-dimensional stereo brightfield microscopy
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