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Confocal microscopy is a standard approach for obtaining volumetric images of a sample with high axial and lateral resolution, especially when dealing with scattering samples. Unfortunately, a confocal microscope is quite expensive compared to traditional microscopes. In addition, the point scanning in confocal microscopy leads to slow imaging speed and photobleaching due to the high dose of laser energy. In this paper, we demonstrate how the advances in machine learning can be exploited to teach a traditional wide-field microscope, one that’s available in every lab, into producing 3D volumetric images like a confocal microscope. The key idea is to obtain multiple images with different focus settings using a wide-field microscope and use a 3D generative adversarial network (GAN) based neural network to learn the mapping between the blurry low-contrast image stacks obtained using a wide-field microscope and the sharp, high-contrast image stacks obtained using a confocal microscope. After training the network with widefield-confocal stack pairs, the network can reliably and accurately reconstruct 3D volumetric images that rival confocal images in terms of its lateral resolution, z-sectioning and image contrast. Our experimental results demonstrate generalization ability to handle unseen data, stability in the reconstruction results, high spatial resolution even when imaging thick (∼40 microns) highly-scattering samples. We believe that such learning-based microscopes have the potential to bring confocal imaging quality to every lab that has a wide-field microscope.
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Design & evaluation of a confocal scanning microscope using off-of-shelf optical components by means of Zemax OpticStudio optical design software
Confocal microscopes are known for optically removing the out-of-focus information (i.e., blur) in each transverse section of the sample’s volume, providing a more accurate three-dimensional image of thick microscopic samples than widefield microscopes. In this work, we have designed and evaluated a confocal microscope using off-of-shelf optical components from Thorlabs’ catalog, one of the major optical manufacturers. The design and evaluation have been implemented using Zemax OpticStudio, the standard optical system design software for realistic optical simulation. Here, we have also reported a practical protocol for building a confocal microscope using sequential mode.
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- Award ID(s):
- 2042563
- PAR ID:
- 10487530
- Editor(s):
- Dr. Sage Graham
- Publisher / Repository:
- QuaesitUM The Undergraduate Research Journal of the University of Memphis
- Date Published:
- Journal Name:
- QuaesitUM The Undergraduate Research Journal of the University of Memphis
- Volume:
- 1
- ISSN:
- 2375-5423
- Page Range / eLocation ID:
- 3-14
- Format(s):
- Medium: X
- Sponsoring Org:
- National Science Foundation
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Accepted Manuscript1.0
- Journal Article:
- The DOI is not currently available.
