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

描述（由申请人提供）：乳腺癌是美国女性癌症相关死亡的第二大原因，也是第二大最常诊断的癌症。放射保乳手术已成为治疗乳腺癌的首选方法，同时保留乳房组织，不影响生存率。乳房部分切除术后局部复发与手术切缘密切相关2-4，但尽管采用术中检查和术后组织病理学相结合的方法进行评估，仍有40%的患者因切缘阳性或闭合5,6而需要再次手术切除。双光子显微镜（TPM）在成像体内和离体内源性和外源性对比方面，比传统的白光和荧光显微镜具有显著的分辨率、分子特异性和光效优势7-9。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 <m10-13。光学相干断层扫描（OCT）是一种利用低相干干涉测量来检测穿透深度为~1-2 mm14的组织散射特性的方法。当与共聚焦显微镜（光学相干显微镜，OCM）结合使用时，OCT的高检测灵敏度和离焦光的相干门控也被证明可以增强通过高散射介质的光学切片。OCM最近通过提供刚切除的大块乳腺病理标本的细胞分辨率图像，证明了光学活检的能力。目前的建议强调波前校正多模态OCM-TPM系统的发展，以快速评估乳腺肿瘤边缘。OCM-TPM将结合各自模式的互补对比度，深度穿透和分子灵敏度的优势。一种新颖的10fs脉冲激光源将用于增强双光子荧光，并允许使用OCM进行精细的轴向切片分辨率。此外，利用自适应校正和反馈算法对散射波前进行补偿，以提高散射组织中OCM和TPM的信号强度和分辨率。同时，我们建议确定最佳的内在和外在生物标志物和替代品，用于乳腺肿瘤边缘的光学评估，包括核的大小和形状，胶原蛋白取向，以及对细胞角蛋白，激素受体和基因扩增敏感的外源性对比。相对于标准免疫组织化学，OCM-TPM的效用将在临床环境中评估，以表征癌症检测的敏感性和特异性。提出的多模态非破坏性光学成像工具将允许术中快速评估手术边缘，作为降低与乳腺癌相关的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.