通过单分子计数和强度校准扩大光测试的动态范围
Lucas Smith, Manish Kohli1, Andrew M Smith
1Department of Oncology , Mayo Clinic , Rochester , Minnesota 55905 , United States.
Journal of the American Chemical Society
|September 15, 2018
概括
这项研究使用成像和分子计数来扩大光测试的动态范围. 它通过克服传统光测量方法的局限性来实现癌症生物标志物的女性分子检测极限.
科学领域:
- 生物化学
- 分析化学
- 生物技术
背景情况:
- 传统的光测试具有有限的动态范围 (1000倍) 和灵敏度 (亚纳米).
- 生物分子表现出大量的变化 (1,000,000倍) 到女性分子水平.
- 现有的方法难以在如此广泛的度范围内准确量化分析物.
研究的目的:
- 扩大基于光的测试的动态范围和灵敏度.
- 开发一种将分子计数与单分子校准结合在一起的方法.
- 克服当前表面捕获试验检测低丰度分析物的局限性.
主要方法:
- 使用成像来结合分子计数与单分子校准组合强度.
- 评估表面捕获的光标签的光学极限,并比较光剂类.
- 使用探测器获取参数来跨越广泛的光辐射范围.
- 使用滚动圆放大用于与癌症相关的miRNA生物标记.
主要成果:
- 鉴定出 phycoerythrin 是一个优越的光体,因为它具有强烈和均的单一光体亮度.
- 在近5个数量级的度范围内实现了精确的单分子检测和校准.
- 对于miRNA生物标志物的检测极限为29个女性分子 (fM).
- 克服了非特定结合的光体所带来的局限性.
结论:
- 开发了成像和分子计数策略,以显著提高光检测性能.
- 展示了扩大表面光测试检测和动态范围的实用方法.
- 该方法可用于在各种生物样本中检测敏感的生物标志物.
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