Superpenetration optical microscopy by iterative multiphoton adaptive compensation technique
Superpenetration optical microscopy by iterative multiphoton adaptive compensation technique
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DOI：
10.1073/pnas.1119590109
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发表时间：
2012-05-29
影响因子：
11.1
通讯作者：
Cui, Meng
中科院分区：
文献类型：
--
作者：
Tang, Jianyong;Germain, Ronald N.;Cui, Meng
生物组织很少是透明的，这对深层组织光学显微镜提出了重大挑战。可实现的成像深度从根本上受到由像差和随机散射引起的波前失真的限制。在这里，我们报告了一种迭代波前补偿技术，利用多光子信号的非线性来确定和补偿这些失真，并将光聚焦在深层组织内。与传统的自适应光学方法不同，该技术可以快速测量深度组织成像中遇到的高度复杂的波前畸变，并提供补偿，不仅像差，但随机散射。该技术是测试与各种高度异质性的生物样品，包括小鼠脑组织，头骨，淋巴结。我们表明，高品质的三维成像可以实现在深度超出了传统的多光子显微镜和自适应光学方法，虽然在有限的距离为一个给定的校正。此外，所需的激光激发功率可以在深部组织中大大降低，偏离弹道光激发的功率要求，从而显著降低对生物组织的光损伤。
Biological tissues are rarely transparent, presenting major challenges for deep tissue optical microscopy. The achievable imaging depth is fundamentally limited by wavefront distortions caused by aberration and random scattering. Here, we report an iterative wavefront compensation technique that takes advantage of the nonlinearity of multiphoton signals to determine and compensate for these distortions and to focus light inside deep tissues. Different from conventional adaptive optics methods, this technique can rapidly measure highly complicated wavefront distortions encountered in deep tissue imaging and provide compensations for not only aberration but random scattering. The technique is tested with a variety of highly heterogeneous biological samples including mouse brain tissue, skull, and lymph nodes. We show that high quality three-dimensional imaging can be realized at depths beyond the reach of conventional multiphoton microscopy and adaptive optics methods, albeit over restricted distances for a given correction. Moreover, the required laser excitation power can be greatly reduced in deep tissues, deviating from the power requirement of ballistic light excitation and thus significantly reducing photo damage to the biological tissue.