温和的无标签的非线性光学成像放松了线性吸收介导三元组
Geng Wang1,2, Lianhuang Li3, Janet E Sorrells2
1Department of Electrical and Computer Engineering, University of Illinois at Urbana-Champaign, Urbana, IL, 61801, USA.
概括
强烈的激光激发会通过线性吸收引起光毒性,而不是非线性吸收. 一种新的快速扫描技术减轻了这种损伤,使得活样本的温和,无标签的分子成像成为可能.
科学领域:
- 生物光子学 生物光子学
- 显微镜的使用方法
- 细胞成像 细胞成像
背景情况:
- 活细胞显微镜需要温和的样本准备,以保持细胞健康.
- 光板显微镜使用平面激发来减少光损伤.
- 激光扫描非线性光学显微镜 (LS-NLM) 尚未实现广泛采用温和,无标签的分子成像,尽管激发受限.
研究的目的:
- 为了研究激光扫描非线性光学显微镜中的光毒性机制.
- 提出和验证一种在无标签成像中减轻光毒性的方法.
- 为了能够对活生物样本进行温和,实时的分子成像.
主要方法:
- 开发了一种光毒性测试,使用母组织模型中的时差自光升高 (超光).
- 在激光扫描非线性光学显微镜中强烈的近红外激发引起的研究光毒性.
- 实施了快速扫描技术,激发时间低于80 femtosecond,完全放松.
主要成果:
- 提供了强有力的证据,证明光毒性源于内在生物分子的线性吸收和随后的三重堆积,而不是非线性吸收.
- 证明,拟议的快速扫描技术有效地减轻了不同样本类型的线性吸收介导光毒性.
- 在高辐射水平下实现了自由移动的Caenorhabditis elegans (C. elegans) 的实时无标签成像.
结论:
- 线性吸收是激光扫描非线性光学显微镜中光毒性的主要驱动因素.
- 一种简单,快速的扫描成像技术可以克服光毒性限制.
- 这种方法可以轻松地进行无标签的分子成像,用于包括C. elegans在内的活生物样品.
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