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相关概念视频

Immunofluorescence Microscopy01:12

Immunofluorescence Microscopy

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A fluorescence microscope uses fluorescent chromophores called fluorochromes, which can absorb energy from a light source and then emit this energy as visible light. Fluorochromes include naturally fluorescent substances (such as chlorophylls) and fluorescent stains that are added to the specimen to create contrast. Dyes such as Texas red and FITC are examples of fluorochromes. Other examples include the nucleic acid dyes 4’,6’-diamidino-2-phenylindole (DAPI), and acridine orange.
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相关实验视频

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A Rapid Method for Multispectral Fluorescence Imaging of Frozen Tissue Sections
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一种用于多重免疫光染色FFPE组织部分的修改过的漂白方法.

Dan Wang1, Alison Cheung1, Gordon E Mawdsley1

  • 1Biomarker Imaging Research Lab, Sunnybrook Research Institute.

Applied immunohistochemistry & molecular morphology : AIMM
|October 7, 2024
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概括

一种经过修改的光化学漂白方法通过改善光体去除和保持组织完整性来增强多重免疫光 (mIF) 染色. 这种技术减少了处理时间,并保持了生物标志物同定位研究的抗原性.

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Author Spotlight: Multiplex Immunofluorescence Combined with Spatial Image Analysis for the Clinical and Biological Assessment of the Tumor Microenvironment
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科学领域:

  • 生物技术是生物技术.
  • 历史学 历史学 历史学
  • 免疫组织化学 免疫组织化学

背景情况:

  • 多重体免疫光 (mIF) 对于生物标志物分析和分析组织部分内的生物标志物相关性至关重要.
  • 标准的Cell DIVE mIF平台使用性过氧化进行顺序的化物失活,但面临着诸如低于最佳的漂白,组织降解和抗原性丧失等挑战.
  • 这些局限性阻碍了高效和可靠的mIF分析,特别是在FPE组织中.

研究的目的:

  • 开发和评估一种经过修改的光化学漂白技术,以克服标准mIF漂白协议的局限性.
  • 使用修改方法与传统方法相比,评估光体去除效率,组织完整性和抗原免疫性.
  • 确定修改技术对整体处理时间的影响及其对多轮染色和成像的实用性.

主要方法:

  • 开发了一种经过修改的光化学漂白方法,涉及强烈的LED光暴露与过氧化应用相结合.
  • 这种技术在乳房和其他组织部位的反复染色/漂白周期中被应用.
  • 剩余光信号,组织完整性和抗原性得到了定量和定性评估,并与传统漂白方法进行了比较.

主要成果:

  • 修改后的光化学漂白有效地消除了前几轮染色的光信号,在多个周期中产生一致的结果.
  • 使用修改方法的光化学处理在整个mIF过程中保留了亚细胞结构和组织抗原性.
  • 八轮mIF手术的处理时间从36小时缩短到30小时,与传统方法相比,组织降解明显减少.

结论:

  • 修改后的光化学漂白技术在mIF中提供可靠的化物去除,显著改善组织完整性和抗原保存.
  • 这种增强的方法减少了整体处理时间,使其成为研究各种组织中生物标志物的共同定位的宝贵工具.
  • 该技术促进了更强大,更有效的多重生物标记分析,克服了传统mIF协议的关键局限性.