Structured Illumination Computational Microscopy with UV Surface Excitation (MUSE) for Multispectral Super-Resolution Histology

用于多光谱超分辨率组织学的紫外表面激发 (MUSE) 结构照明计算显微镜

基本信息

  • 批准号:
    10213544
  • 负责人:
  • 金额:
    $ 2.31万
  • 依托单位:
  • 依托单位国家:
    美国
  • 项目类别:
  • 财政年份:
    2018
  • 资助国家:
    美国
  • 起止时间:
    2018-09-01 至 2020-12-31
  • 项目状态:
    已结题

项目摘要

Project Summary/Abstract Current clinical practices for the diagnosis and management of diseases often rely on histopathological exami- nation of tissue via optical microscopy. Brightfield imaging of hematoxylin-eosin (H&E)-stained samples repre- sents the predominant approach for accurate and comprehensive evaluation and diagnosis in clinical histopathol- ogy [1, 2]. Additional techniques for disease characterization involve molecularly specific labeling, and use im- munohistochemical or immunofluorescence techniques for brightfield and fluorescence microscopy, respec- tively. Using the latter, multiple analytes can be examined simultaneously [3, 4]. Unfortunately, the complexity of a fluorescent microscope’s optical design scales with the number of multiplexed fluorescent reporters to visual- ize, thus limiting its clinical utility [5]. Another area of interest is to explore clinically relevant information that may exist at spatial resolutions beyond what can be achieved with conventional microscopes. Typical fluorescence microscopy is generally limited by diffraction to an optical resolution of ~200 nm. Though this resolution enables visualization of large cellular structures, it does not support examination of organelle- and suborganelle-level ultrastructure whose morphological changes can correlate with disease, as seen in neurodegeneration, age, and cancer [6-10]. Recently, optical super-resolution technologies have been introduced that achieve imaging reso- lutions better than 50 nm. However, such technologies depend on complex hardware and are currently too costly to be incorporated into typical clinical pathology budgets. Electron microscopy (EM) systems are also an availa- ble option, and routinely image at resolutions of ~1 nm – however, these are not widely available and are not well suited for molecular specific imaging [11-14]. Additional issues, including size, cost, limited field-of-view, and complexity of sample-prep protocols have prevented EM from being incorporated into standard clinical work- flow. This project will develop a robust, comparatively simple, and low-cost optical system for molecularly-specific multispectral fluorescence imaging at spatial resolutions of ~70 nm, well beneath the classical 200 nm optical resolution limit. To do so, a framework for computational structured illumination (SI) microscopy will be developed to enable super-resolution using uncalibrated illumination patterns. This framework will be deployed using single- wavelength ultraviolet (UV) excitation, which has demonstrated capabilities for simultaneous excitation of multi- ple fluorescent reporters. Specific innovations in this work include a novel reformulation of SI microscopy that uses computational optimization to robustly increase imaging resolution in the presence of system unknowns and imperfections. Furthermore, because UV-based excitation has wavelengths more than a factor of 2 shorter than the fluorophores’ visible emission wavelengths, resolution gains by factors greater than 2 are expected, hence enabling sub-100-nm spatial resolutions. If successful, the aims of this project will combine the benefits of multispectral optical imaging with the advantages of sub-100-nm spatial resolution to create a more informative and less demanding alternative to electron microscopy, with applications across biology and histopathology.
项目总结/文摘

项目成果

期刊论文数量(2)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
Computational structured illumination for high-content fluorescence and phase microscopy.
  • DOI:
    10.1364/boe.10.001978
  • 发表时间:
    2018-12
  • 期刊:
  • 影响因子:
    3.4
  • 作者:
    Li-Hao Yeh;Shwetadwip Chowdhury;L. Waller
  • 通讯作者:
    Li-Hao Yeh;Shwetadwip Chowdhury;L. Waller
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Shwetadwip Chowdhury其他文献

Shwetadwip Chowdhury的其他文献

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{{ truncateString('Shwetadwip Chowdhury', 18)}}的其他基金

Computational Framework to Enhance Antenna-based Electromagnetic Imaging
增强基于天线的电磁成像的计算框架
  • 批准号:
    10667975
  • 财政年份:
    2023
  • 资助金额:
    $ 2.31万
  • 项目类别:
Structured Illumination Computational Microscopy with UV Surface Excitation (MUSE) for Multispectral Super-Resolution Histology
用于多光谱超分辨率组织学的紫外表面激发 (MUSE) 结构照明计算显微镜
  • 批准号:
    9788760
  • 财政年份:
    2018
  • 资助金额:
    $ 2.31万
  • 项目类别:

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