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Imaging Biological Samples with Optical Microscopy01:18

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Optical microscopy uses optic principles to provide detailed images of samples. Antonie van Leeuwenhoek designed the first compound optical microscope in the 17th century to visualize blood cells, bacteria, and yeast cells. In 1830, Joseph Jackson Lister created an essentially modern light microscope. The 20th century saw the development of microscopes with enhanced magnification and resolution.
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在多光谱显微镜之间比较和纠正光谱敏感性:临床实施的先决条件.

Margaret Eminizer1,2, Melinda Nagy1,2, Elizabeth L Engle3,4,5

  • 1Department of Physics and Astronomy, Johns Hopkins University, Baltimore, MD 21210, USA.

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概括

标准化多谱,多重免疫光 (mIF) 显微镜对于可重复诊断至关重要. 一个简单的校准模型显著减少了不同显微镜的图像变化,增强了临床病理学的应用.

关键词:
校准校准的时间高吞吐量,具有高吞吐量.影像成像技术 影像成像技术免疫光显微镜的显微镜.病理学的病理学系统学是一种系统学.

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科学领域:

  • 生物医学成像技术 生物医学成像技术
  • 病理学 病理学 病理学
  • 计算生物学 计算生物学

背景情况:

  • 多光谱,多重免疫光 (mIF) 显微镜能够在组织中进行详细的细胞分析.
  • 在不同的显微镜中,mIF图像数据的可复制性和标准化对于诊断应用至关重要.
  • 当前的mIF技术需要强大的方法来确保一致的数据质量.

研究的目的:

  • 描述和纠正不同mIF显微镜之间照明强度和光谱灵敏度的变化.
  • 开发和验证用于标准化mIF图像数据的校准模型.
  • 评估标准化对减少图像变化和提高标记表达精度的影响.

主要方法:

  • 在三个不同的mIF显微镜上扫描了8个黑色素瘤组织样本.
  • 计算平均组织区域流量强度以确定基线变化.
  • 应用特定于样品,特定于显微镜的校准模型来纠正亮度和光谱差异.
  • 在独立样本子集上验证校准模型.

主要成果:

  • 显微镜的基线平均标准偏差为29.9%,在初始校正后降至13.9%.
  • 校准模型的应用使验证子集的变化减少到2.9 ± 0.03%.
  • 在未混合标记物表达的变化在经过校正后从15.8%降至4.4%.

结论:

  • 一个简单的校准模型可以有效地标准化mIF显微镜.
  • 标准化显著减少了图像数据的变化,提高了可重现性.
  • 标准化mIF显微镜对于临床病理学实验室来说是可行的.