Deep learning extended depth-of-field microscope for fast and slide-free histology

Microscopic evaluation of resected tissue plays a central role in the surgical management of cancer. Because optical microscopes have a limited depth-of-field (DOF), resected tissue is either frozen or preserved with chemical fixatives, sliced into thin sections placed on microscope slides, stained,...

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Veröffentlicht in:Proceedings of the National Academy of Sciences - PNAS Jg. 117; H. 52; S. 33051
Hauptverfasser: Jin, Lingbo, Tang, Yubo, Wu, Yicheng, Coole, Jackson B, Tan, Melody T, Zhao, Xuan, Badaoui, Hawraa, Robinson, Jacob T, Williams, Michelle D, Gillenwater, Ann M, Richards-Kortum, Rebecca R, Veeraraghavan, Ashok
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Sprache:Englisch
Veröffentlicht: United States 29.12.2020
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ISSN:1091-6490, 1091-6490
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Abstract Microscopic evaluation of resected tissue plays a central role in the surgical management of cancer. Because optical microscopes have a limited depth-of-field (DOF), resected tissue is either frozen or preserved with chemical fixatives, sliced into thin sections placed on microscope slides, stained, and imaged to determine whether surgical margins are free of tumor cells-a costly and time- and labor-intensive procedure. Here, we introduce a deep-learning extended DOF (DeepDOF) microscope to quickly image large areas of freshly resected tissue to provide histologic-quality images of surgical margins without physical sectioning. The DeepDOF microscope consists of a conventional fluorescence microscope with the simple addition of an inexpensive (less than $10) phase mask inserted in the pupil plane to encode the light field and enhance the depth-invariance of the point-spread function. When used with a jointly optimized image-reconstruction algorithm, diffraction-limited optical performance to resolve subcellular features can be maintained while significantly extending the DOF (200 µm). Data from resected oral surgical specimens show that the DeepDOF microscope can consistently visualize nuclear morphology and other important diagnostic features across highly irregular resected tissue surfaces without serial refocusing. With the capability to quickly scan intact samples with subcellular detail, the DeepDOF microscope can improve tissue sampling during intraoperative tumor-margin assessment, while offering an affordable tool to provide histological information from resected tissue specimens in resource-limited settings.
AbstractList Microscopic evaluation of resected tissue plays a central role in the surgical management of cancer. Because optical microscopes have a limited depth-of-field (DOF), resected tissue is either frozen or preserved with chemical fixatives, sliced into thin sections placed on microscope slides, stained, and imaged to determine whether surgical margins are free of tumor cells-a costly and time- and labor-intensive procedure. Here, we introduce a deep-learning extended DOF (DeepDOF) microscope to quickly image large areas of freshly resected tissue to provide histologic-quality images of surgical margins without physical sectioning. The DeepDOF microscope consists of a conventional fluorescence microscope with the simple addition of an inexpensive (less than $10) phase mask inserted in the pupil plane to encode the light field and enhance the depth-invariance of the point-spread function. When used with a jointly optimized image-reconstruction algorithm, diffraction-limited optical performance to resolve subcellular features can be maintained while significantly extending the DOF (200 µm). Data from resected oral surgical specimens show that the DeepDOF microscope can consistently visualize nuclear morphology and other important diagnostic features across highly irregular resected tissue surfaces without serial refocusing. With the capability to quickly scan intact samples with subcellular detail, the DeepDOF microscope can improve tissue sampling during intraoperative tumor-margin assessment, while offering an affordable tool to provide histological information from resected tissue specimens in resource-limited settings.
Microscopic evaluation of resected tissue plays a central role in the surgical management of cancer. Because optical microscopes have a limited depth-of-field (DOF), resected tissue is either frozen or preserved with chemical fixatives, sliced into thin sections placed on microscope slides, stained, and imaged to determine whether surgical margins are free of tumor cells-a costly and time- and labor-intensive procedure. Here, we introduce a deep-learning extended DOF (DeepDOF) microscope to quickly image large areas of freshly resected tissue to provide histologic-quality images of surgical margins without physical sectioning. The DeepDOF microscope consists of a conventional fluorescence microscope with the simple addition of an inexpensive (less than $10) phase mask inserted in the pupil plane to encode the light field and enhance the depth-invariance of the point-spread function. When used with a jointly optimized image-reconstruction algorithm, diffraction-limited optical performance to resolve subcellular features can be maintained while significantly extending the DOF (200 µm). Data from resected oral surgical specimens show that the DeepDOF microscope can consistently visualize nuclear morphology and other important diagnostic features across highly irregular resected tissue surfaces without serial refocusing. With the capability to quickly scan intact samples with subcellular detail, the DeepDOF microscope can improve tissue sampling during intraoperative tumor-margin assessment, while offering an affordable tool to provide histological information from resected tissue specimens in resource-limited settings.Microscopic evaluation of resected tissue plays a central role in the surgical management of cancer. Because optical microscopes have a limited depth-of-field (DOF), resected tissue is either frozen or preserved with chemical fixatives, sliced into thin sections placed on microscope slides, stained, and imaged to determine whether surgical margins are free of tumor cells-a costly and time- and labor-intensive procedure. Here, we introduce a deep-learning extended DOF (DeepDOF) microscope to quickly image large areas of freshly resected tissue to provide histologic-quality images of surgical margins without physical sectioning. The DeepDOF microscope consists of a conventional fluorescence microscope with the simple addition of an inexpensive (less than $10) phase mask inserted in the pupil plane to encode the light field and enhance the depth-invariance of the point-spread function. When used with a jointly optimized image-reconstruction algorithm, diffraction-limited optical performance to resolve subcellular features can be maintained while significantly extending the DOF (200 µm). Data from resected oral surgical specimens show that the DeepDOF microscope can consistently visualize nuclear morphology and other important diagnostic features across highly irregular resected tissue surfaces without serial refocusing. With the capability to quickly scan intact samples with subcellular detail, the DeepDOF microscope can improve tissue sampling during intraoperative tumor-margin assessment, while offering an affordable tool to provide histological information from resected tissue specimens in resource-limited settings.
Author Coole, Jackson B
Williams, Michelle D
Badaoui, Hawraa
Robinson, Jacob T
Gillenwater, Ann M
Wu, Yicheng
Zhao, Xuan
Richards-Kortum, Rebecca R
Jin, Lingbo
Tang, Yubo
Tan, Melody T
Veeraraghavan, Ashok
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  organization: Department of Bioengineering, Rice University, Houston, TX 77005
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  organization: Department of Applied Physics, Rice University, Houston, TX 77005
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  orcidid: 0000-0002-8821-4139
  surname: Coole
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  organization: Department of Bioengineering, Rice University, Houston, TX 77005
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  surname: Tan
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  organization: Department of Bioengineering, Rice University, Houston, TX 77005
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  organization: Department of Applied Physics, Rice University, Houston, TX 77005
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  organization: Department of Head and Neck Surgery, University of Texas MD Anderson Cancer Center, Houston, TX 77030
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  fullname: Robinson, Jacob T
  organization: Department of Bioengineering, Rice University, Houston, TX 77005
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  givenname: Michelle D
  surname: Williams
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  organization: Department of Pathology, University of Texas MD Anderson Cancer Center, Houston, TX 77030
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  surname: Gillenwater
  fullname: Gillenwater, Ann M
  organization: Department of Head and Neck Surgery, University of Texas MD Anderson Cancer Center, Houston, TX 77030
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  orcidid: 0000-0003-2347-9467
  surname: Richards-Kortum
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  organization: Department of Bioengineering, Rice University, Houston, TX 77005; rkortum@rice.edu vashok@rice.edu
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  givenname: Ashok
  orcidid: 0000-0001-5043-7460
  surname: Veeraraghavan
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  email: rkortum@rice.edu, vashok@rice.edu
  organization: Department of Electrical and Computer Engineering, Rice University, Houston, TX 77005; rkortum@rice.edu vashok@rice.edu
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Keywords deep learning
end-to-end optimization
pathology
phase mask
extended depth-of-field microscopy
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Snippet Microscopic evaluation of resected tissue plays a central role in the surgical management of cancer. Because optical microscopes have a limited depth-of-field...
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SubjectTerms Algorithms
Animals
Biopsy - instrumentation
Biopsy - methods
Biopsy - standards
Calibration
Carcinoma - pathology
Deep Learning
Humans
Image Processing, Computer-Assisted - instrumentation
Image Processing, Computer-Assisted - methods
Image Processing, Computer-Assisted - standards
Microscopy, Fluorescence - instrumentation
Microscopy, Fluorescence - methods
Microscopy, Fluorescence - standards
Mouth Neoplasms - pathology
Swine
Title Deep learning extended depth-of-field microscope for fast and slide-free histology
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