Feasibility study of Raman spectroscopy for investigating the mouse retina in vivo
Feasibility study of Raman spectroscopy for investigating the mouse retina in vivo
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拉曼光谱用于体内研究小鼠视网膜的可行性研究
DOI：
10.1117/12.2288790
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发表时间：
2018
期刊：
影响因子：
--
通讯作者：
Söderberg, Per G.
中科院分区：
文献类型：
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作者：
Zhang, Pengfei;Maleppat, Ratheesh K.;Pugh, Edward N.;de Oliveira, Marcos A.;Chang, Che-Wei;Chan, James W.;Zawadzki, Robert J.;Manna, Suman K.;Manns, Fabrice;Söderberg, Per G.
拉曼光谱在生物化学中的应用非常成功，特别是因为它能够识别基本的化学物种。然而，这种光谱技术在体内评估中的应用往往受到自体荧光的限制，这使得拉曼信号的检测变得困难。小鼠的眼睛已经被用作研究各种疾病模型和治疗途径的光学试验台。在小鼠视网膜中实现体内拉曼光谱将是有价值的，但需要在使用高探测光束功率时固有的自动荧光伪影和潜在的光损伤的背景下进行检查。为了评估可行性，在定制的SLO/OCT平台上建立了一个拉曼系统，允许鼠标定位和形态数据采集，以及来自所需视网膜偏心率的拉曼信号。首先在成像平面(视网膜)中使用由聚苯乙烯组成的模型眼来评估拉曼系统的性能，使用488 nm、561 nm和785 nm的激发波长来确定在较长波长下自发荧光是否会减弱。为了提高信噪比，组合系统具有对这三种激发的光学兼容性，以便在单次成像过程中可以连续获取典型感兴趣区域的相应光谱。我们的结果包括在10多个激发能小于160J.S~(-1)·m~(-2)的S对所有波长的发射光谱以及不同品系包括WT、BALB/c和ABCA4-/-的视网膜形态。
The use of Raman spectroscopy in biochemistry has been very successful, particularly because of its ability to identify elementary chemical species. However, application of this spectroscopic technique for in vivo assessment is often limited by autofluorescence, which make detection of Raman signatures difficult. The mouse eye has been used as an optical testbed for investigation of a variety of disease models and therapeutic pathways. Implementation of in vivo Raman spectroscopy in mice retina would be valuable but needs to be examined in context of the intrinsic auto-fluorescence artifact and potential light damage if high probing beam powers were used. To evaluate feasibility, a Raman system was built on a custom SLO/OCT platform allowing mouse positioning and morphological data acquisition along with the Raman signal from a desired retinal eccentricity. The performance of the Raman system was first assessed with a model eye consisting of polystyrene in the image plane (retina), using excitation wavelengths of 488 nm, 561 nm, and 785 nm to determine whether auto-fluorescence would be reduced at longer wavelengths. To improve the SNR, the combined system is featured with the optical compatibility for these three excitations such that their corresponding spectra from a typical region of interest can be acquired consecutively during single imaging run. Our results include emission spectra acquired over 10 s with excitation energy less than 160 J.s-1.m-2for all wavelengths and corresponding retinal morphology for different mouse strains including WT, BALB/c and ABCA4-/-.
生物动力学中的时间分辨振动光谱
DOI：
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发表时间：
1994
期刊：
影响因子：
--
作者：
W. Woodruff
通讯作者：
W. Woodruff