Multiphoton microscopy of prostate and periprostatic neural tissue: a promising imaging technique for improving nerve-sparing prostatectomy.
Multiphoton microscopy of prostate and periprostatic neural tissue: a promising imaging technique for improving nerve-sparing prostatectomy.
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前列腺和前列腺周围神经组织的多光子显微镜：一种有前途的成像技术，用于改善保留神经的前列腺切除术。
DOI：
10.1089/end.2009.0221
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发表时间：
2009
影响因子：
2.7
通讯作者：
Tewari,AshutoshK
中科院分区：
文献类型：
--
作者：
Yadav,Rajiv;Mukherjee,Sushmita;Hermen,Michael;Tan,Gerald;Maxfield,FrederickR;Webb,WattW;Tewari,AshutoshK
背景和目的：各种成像模式正在研究中的实时组织成像前列腺周围神经的想法，以提高保留神经的根治性前列腺切除术的结果。我们探讨了多光子显微镜（MPM）的实时组织成像的前列腺和前列腺周围的神经组织在雄性Sprague-Dawley大鼠模型。这种技术的独特优势是高分辨率图像的采集，而不需要任何外在的标记剂和最小的光毒性效应对tissue.Materials和方法：前列腺和海绵体神经手术切除雄性Sprague-Dawley大鼠。使用组织的固有荧光和散射特性进行成像，而没有任何外源性染料或造影剂。使用定制的MPM，由Olympus BX 61 WI直立框架和改良的MRC 1024扫描头组成。使用波长为780 nm的飞秒脉冲钛/蓝宝石激光激发组织;通过普克尔斯盒调制物镜下的激光功率。在390（±35 nm）处收集二次谐波产生（SHG）信号，并在380至530 nm处收集宽带自体荧光。然后将从SHG和组织荧光获得的图像合并，并在后处理期间进行颜色编码，以更好地欣赏细节。相应的组织进行苏木精和伊红染色的结构的组织学confirmations.Results：高分辨率图像的前列腺包膜，下面的腺泡，和个别细胞概述的腺体在不同的放大倍数。脂肪组织和神经组织的MPM图像也被获得。前列腺、脂肪、海绵体神经和盆大神经节的组织学确认和相关性验证了MPM的发现。结论：使用MPM的前列腺和前列腺周围神经组织的实时成像和显微分辨率是可行的，而不需要任何外部标记剂。这种成像方式与手术技术的整合有可能提高保留神经的脊髓束切除术的精确度。
Background and Purpose:Various imaging modalities are under investigation for real-time tissue imaging of periprostatic nerves with the idea of improving the results of nerve-sparing radical prostatectomy. We explored multiphoton microscopy (MPM) for real-time tissue imaging of the prostate and periprostatic neural tissue in a male Sprague-Dawley rat model. The unique advantage of this technique is the acquisition of high-resolution images without necessitating any extrinsic labeling agent and with minimal phototoxic effect on tissue.Materials and Methods:The prostate and cavernous nerves were surgically excised from male Sprague-Dawley rats. The imaging was carried out using intrinsic fluorescence and scattering properties of the tissues without any exogenous dye or contrast agent. A custom-built MPM, consisting of an Olympus BX61WI upright frame and a modified MRC 1024 scanhead, was used. A femtosecond pulsed titanium/sapphire laser at 780-nm wavelength was used to excite the tissue; laser power under the objective was modulated via a Pockels cell. Second harmonic generation (SHG) signals were collected at 390 (±35 nm), and broadband autofluorescence was collected at 380 to 530 nm. The images obtained from SHG and from tissue fluorescence were then merged and color coded during postprocessing for better appreciation of details. The corresponding tissues were subjected to hematoxylin and eosin staining for histologic confirmation of the structures.Results:High-resolution images of the prostate capsule, underlying acini, and individual cells outlining the glands were obtained at varying magnifications. MPM images of adipose tissue and the neural tissues were also obtained. Histologic confirmation and correlation of the prostate gland, fat, cavernous nerve, and major pelvic ganglion validated the findings of MPM.Conclusion:Real-time imaging and microscopic resolution of prostate and periprostatic neural tissue using MPM is feasible without the need for any extrinsic labeling agents. Integration of this imaging modality with operative technique has the potential to improve the precision of nerve-sparing prostatectomy.