Recent advances in nonlinear microscopy: Deep insights and polarized revelations.

Recent advances in nonlinear microscopy: Deep insights and polarized revelations.
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DOI:
10.1016/j.biocel.2020.105896
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发表时间:
2021-01
期刊:
The international journal of biochemistry & cell biology
影响因子:
--
通讯作者:
Wiseman PW
Wiseman PW
中科院分区:
其他
文献类型:
--
作者:
Gopal AA;Kazarine A;Dubach JM;Wiseman PW

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非线性显微镜是一种利用光与物质之间的非线性相互作用来成像生物组织中的荧光和散射现象的技术。非线性激发需要聚焦短脉冲激光的非常高的峰值强度,因为同时到达激发荧光团或与选择性结构相互作用以产生谐波的低能量的多个光子的概率极低。结合较长波的利用减少了散射,固有的空间限制与在焦体积内实现光子的同时到达有关,使非线性成像具有低离焦背景的深度成像。这篇综述介绍了不同的对比机制,可与非线性成像和仪器常用的非线性显微镜。此外,我们还讨论了非线性显微镜在扩大成像穿透深度、对新鲜组织进行组织病理学研究以及利用偏振非线性显微镜检测分子的顺序和取向等方面的最新进展。
Nonlinear microscopy is a technique that utilizes nonlinear interactions between light and matter to image fluorescence and scattering phenomena in biological tissues. Very high peak intensities from focused short pulsed lasers are required for nonlinear excitation due to the extremely low probability of the simultaneous arrival of multiple photons of lower energy to excite fluorophores or interact with selective structures for harmonic generation. Combined with reduced scattering from the utilization of longer wavelengths, the inherent spatial confinement associated with achieving simultaneous arrival of photons within the focal volume enables deep imaging with low out-of-focus background for nonlinear imaging. This review provides an introduction to the different contrast mechanisms available with nonlinear imaging and instrumentation commonly used in nonlinear microscopy. Furthermore, we discuss some recent advances in nonlinear microscopy to extend the imaging penetration depth, conduct histopathological investigations on fresh tissues and examine the molecular order and orientation of molecules using polarization nonlinear microscopy.
DOI: 10.1039/c9an00267g
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