Optical Characterization of Neurosurgical Operating Microscopes: Quantitative Fluorescence and Assessment of PpIX Photobleaching.
Optical Characterization of Neurosurgical Operating Microscopes: Quantitative Fluorescence and Assessment of PpIX Photobleaching.
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DOI：
10.1038/s41598-018-30247-6
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发表时间：
2018-08-22
影响因子：
4.6
通讯作者：
Preul MC
中科院分区：
文献类型：
--
作者：
Belykh E;Miller EJ;Patel AA;Bozkurt B;Yağmurlu K;Robinson TR;Nakaji P;Spetzler RF;Lawton MT;Nelson LY;Seibel EJ;Preul MC
5-氨基乙酰丙酸（5-ALA）诱导的原卟啉IX （PpIX）越来越多地被用作荧光引导恶性胶质瘤切除术的荧光标记物。了解激发光源和PpIX荧光如何与手术显微镜相互作用的特性对于有效利用荧光引导的肿瘤切除技术至关重要。在本研究中，我们对用于PpIX可视化的临床级操作显微镜发射的蓝光和白光强度以及光束轮廓进行了详细的评估。这些测量揭示了公认的荧光光漂白局限性和未被认识到的局限性，这些局限性可能会改变使用操作显微镜获得的PpIX荧光定量观察结果，从而对研究和临床应用产生潜在影响。我们还评估了具有类似PpIX的激发-发射轮廓的光稳定荧光标准的光学性质。此外，我们在动物胶质瘤模型中测量了5- ala诱导的PpIX荧光的时间依赖性动力学。最后，我们开发了一种定量PpIX荧光的比率法，该方法使用光稳定的荧光标准物将PpIX荧光强度归一化。该方法提高了准确性，并允许多个样品测量的可重复性和直接可比性。
Protoporphyrin IX (PpIX) induced by 5-aminolevulinic acid (5-ALA) is increasingly used as a fluorescent marker for fluorescence-guided resection of malignant gliomas. Understanding how the properties of the excitation light source and PpIX fluorescence interact with the surgical microscope is critical for effective use of the fluorescence-guided tumor resection technique. In this study, we performed a detailed assessment of the intensity of the emitted blue light and white light and the light beam profile of clinical grade operating microscopes used for PpIX visualization. These measurements revealed both recognized fluorescence photobleaching limitations and unrecognized limitations that may alter quantitative observations of PpIX fluorescence obtained with the operating microscope with potential impact on research and clinical uses. We also evaluated the optical properties of a photostable fluorescent standard with an excitation-emission profile similar to PpIX. In addition, we measured the time-dependent dynamics of 5-ALA-induced PpIX fluorescence in an animal glioma model. Finally, we developed a ratiometric method for quantification of the PpIX fluorescence that uses the photostable fluorescent standard to normalize PpIX fluorescence intensity. This method increases accuracy and allows reproducible and direct comparability of the measurements from multiple samples.
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影响因子：
4.1
作者：
Lau, Darryl;Hervey-Jumper, Shawn L.;Berger, Mitchel S.
通讯作者：
Berger, Mitchel S.
影响因子：
1.8
作者：
Belykh E;Martirosyan NL;Yagmurlu K;Miller EJ;Eschbacher JM;Izadyyazdanabadi M;Bardonova LA;Byvaltsev VA;Nakaji P;Preul MC
通讯作者：
Preul MC
影响因子：
--
作者：
Kim, Hong Rae;Lee, Hyun Min;Kim, Kwang Gi
通讯作者：
Kim, Kwang Gi
影响因子：
28.4
作者：
Brown TJ;Brennan MC;Li M;Church EW;Brandmeir NJ;Rakszawski KL;Patel AS;Rizk EB;Suki D;Sawaya R;Glantz M
通讯作者：
Glantz M
影响因子：
4.6
作者：
Kim SK;Kirchner EA;Stefes A;Kirchner F
通讯作者：
Kirchner F