激光时钟的模拟复合和计算光子计数用于快速光寿命成像显微镜
Rishyashring R Iyer1,2, Janet E Sorrells1,3, Kevin K D Tan1,3
1Beckman Institute for Advanced Science and Technology, University of Illinois Urbana-Champaign, Urbana, IL 61801, USA.
Biomedical optics express
|April 18, 2024
概括
这项研究引入了时间域复杂化,以实现更快,更准确的光终身成像显微镜 (FLIM). 这种新方法改善了对具有挑战性的低光生物样本的寿命估计.
科学领域:
- 生物光子学 生物光子学
- 显微镜的使用方法
- 频谱学是一种光谱学.
背景情况:
- 生物样本在光强度和寿命方面表现出广泛的动态范围.
- 高速光终身成像显微镜 (FLIM) 对于精确的生物分析至关重要.
- 现有的快速FLIM方法与光子缺乏条件和数据效率低下作斗争.
研究的目的:
- 在快速FLIM中开发一种用于准确记录光子到达时间的新方法.
- 为了克服传统FLIM同步和数据处理的局限性.
- 为了提高稀疏或弱光生物样本的寿命估计准确度.
主要方法:
- 实现了时间域复杂化,以记录光子到达时间与激发激光钟.
- 用光电流将激光时钟信号复合到未被占用的时间段中.
- 使用GHz采样速率的计算光子计数.
主要成果:
- 消除了光子饥饿样本的寿命估计中的错误.
- 实现了亚纳秒死亡时间,与现有的快速FLIM方法相提并论.
- 在具有不同激光重复率的多光子显微镜设置上证明有效的单通道和多通道FLIM.
结论:
- 时间域复杂化为具有挑战性的生物成像中快速FLIM提供了强大的解决方案.
- 该技术提高了准确性和效率,而不会增加数据瓶.
- 这一进步使生物样品的实时成像能够以更高的精度进行.
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