Wavefront Corrected Multimodal Optical Coherence and Two-Photon Microscopy for Ra

描述(申请人提供)：乳腺癌是美国女性癌症相关死亡的第二大原因，也是第二常见的确诊癌症1。放射保乳手术已经成为治疗乳腺癌的首选方法，在保留乳腺组织的同时不会影响存活率。乳房部分切除术后的局部复发与手术切缘2-4密切相关，但尽管结合术中检查和术后组织病理学进行评估，40%的患者由于切缘阳性或接近切缘而需要再次手术切除5，6。双光子显微镜(TPM)在体内和体外对比剂成像方面显示出显著的分辨率、分子特异性和光效优势，优于传统的白光和荧光显微镜。然而，散射组织严重降低了双光子信号和分辨率，将TPM的基本成像深度限制在~100&lt；m10-13。光学相干层析成像(OCT)是一种利用低相干度干涉技术来检测穿透深度为~1-2 mm的组织的散射特性的方法。与共焦显微镜(光学相干显微镜，OCM)一起使用时，OCT的高检测灵敏度和相干选通也被证明可以通过高散射介质增强光学切片15。OCM最近通过提供新鲜切除的大块乳腺病理标本的细胞分辨率图像16展示了光学活检的能力。目前的提案强调开发波前校正的多模式OCM-TPM系统来快速评估乳腺肿瘤的边缘。OCM-TPM将结合各自模式的互补对比度、深度渗透和分子敏感性的优势。一种新型的10ps脉冲激光光源将被用来增强双光子荧光，并允许使用OCM进行精确的轴向切片分辨率。此外，还将通过自适应校正和反馈算法来补偿散射波前，以提高散射组织中OCM和TPM的信号强度和分辨率。同时，我们建议找出最佳的内在和外在生物标志物和替代物，用于对乳腺肿瘤边缘进行光学评估，包括核大小、形状和胶原取向，以及对细胞角蛋白、激素受体和基因扩增的外源性对比敏感。相对于标准免疫组织化学，OCM-TPM的实用性将在临床环境中进行评估，以表征癌症检测的敏感性和特异性。拟议的多模式非破坏性光学成像工具将允许在手术中快速评估手术切缘，作为降低乳腺癌相关发病率和死亡率的一种手段。

DESCRIPTION (provided by applicant): Breast cancer is the second leading cause of cancer-related death and second most frequently diagnosed cancer in women in the United States1. Breast conserving surgery with radiation has emerged as the preferred method for treating breast cancer while preserving breast tissue and without compromising survival rates. Local recurrence following partial mastectomy is strongly correlated to the surgical margin2-4, but despite evaluation using a combination of intraoperative examination and post-operative histopathology, 40% of patients require surgical re-excision as a result of positive or close margins5, 6. Two-photon microscopy (TPM) has demonstrated significant resolution, molecular specificity, and light efficiency advantages over conventional white-light and fluorescence microscopes for imaging endogenous and exogenous contrast both in vivo and ex vivo7-9. However, scattering tissue severely degrades two-photon signals and resolution, limiting the fundamental imaging depth of TPM to ~100 <m10-13. Optical coherence tomography (OCT) is a method of utilizing low-coherence interferometry to detect the scattering properties of tissue down to penetration depths of ~1-2 mm14. The high detection sensitivity and coherence-gating of out-of-focus light of OCT has also been shown to enhance optical sectioning through highly scattering media when used in conjunction with confocal microscopy (optical coherence microscopy, OCM)15. OCM has recently demonstrated the capacity for optical biopsy by providing cellular-resolution images of freshly-excised bulk breast pathology specimens16. The current proposal emphasizes the development of a wavefront corrected multimodal OCM-TPM system to rapidly assess breast tumor margins. OCM-TPM will combine the advantages of complimentary contrast, depth penetration, and molecular-sensitivity of the respective modalities. A novel 10 fs pulsed laser source will be used to enhance two-photon fluorescence and allow for exquisite axial sectioning resolution using OCM. Further, the scattering wavefront will be compensated by using adaptive correction and feedback algorithms to improve the signal intensity and resolution of both OCM and TPM in scattering tissue. Concurrently, we propose to identify optimal intrinsic and extrinsic biomarkers and surrogates for optical evaluation of breast tumor margins, including nuclear size and shape and collagen orientation, as well as exogenous contrast sensitive to cytokeratins, hormone receptors, and gene amplification. The utility of OCM-TPM, relative to standard immunohistochemistry, will be assessed in a clinical setting to characterize cancer detection sensitivity and specificity. The proposed multimodal non-destructive optical imaging tool will allow for rapid intraoperative assessment of surgical margins as a means of reducing the morbidity and mortality associated with breast cancer.