Two-color, two-photon, and excited-state absorption microscopy
Two-color, two-photon, and excited-state absorption microscopy
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DOI：
10.1117/1.2780173
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发表时间：
2007-09-01
影响因子：
3.5
通讯作者：
Warren, S. Warren
中科院分区：
文献类型：
--
作者：
Fu, Dan;Ye, Tong;Warren, S. Warren
我们开发了一种利用双色双光子和激发态吸收显微镜成像非荧光物质的新方法。如果两个不同颜色的同步锁模脉冲序列中的一个被强度调制，当介质中发生非线性吸收时，调制转移到另一个脉冲序列。我们可以很容易地测量由双光子吸收或激发态吸收引起的10（-6）吸收变化。本文以黑褐色黑色素为模型系统进行了详细的研究。在溶液中对黑色素的瞬时双光子吸收（TPA）和相对长寿命激发态吸收（ESA）进行了光谱学研究，并在B16细胞中证明了其成像能力。在775 nm的泵浦下，棕褐色黑色素表现出两种不同的激发态，具有不同的寿命（一种为3ps，一种持续数百纳秒）。其特征的TPA/ESA使我们能够以与双光子荧光显微镜（TPFM）相当的高分辨率成像其在细胞样品中的分布。这项新技术可能为诊断黑色素瘤提供有价值的信息。(C) 2007光学仪器工程师学会。
We develop a new approach in imaging nonfluorescent species with two-color two-photon and excited state absorption microscopy. If one of two synchronized mode-locked pulse trains at different colors is intensity modulated, the modulation transfers to the other pulse train when nonlinear absorption takes places in the medium. We can easily measure 10(-6) absorption changes caused by either two-photon absorption or excited-state absorption with a RF lock-in amplifier. Sepia melanin is studied in detail as a model system. Spectroscopy studies on the instantaneous two-photon absorption (TPA) and the relatively long-lived excited-state absorption (ESA) of melanin are carried out in solution, and imaging capability is demonstrated in B16 cells. It is found that sepia melanin exhibits two distinct excited states with different lifetimes (one at 3 ps, one lasting hundreds of nanoseconds) when pumped at 775 nm. Its characteristic TPA/ESA enables us to image its distribution in cell samples with high resolution comparable to two-photon fluorescence microscopy (TPFM). This new technique could potentially provide valuable information in diagnosing melanoma. (C) 2007 Society of Photo-Optical Instrumentation Engineers.