Two-Photo Optical Biopsy Probe

描述（由申请人提供）： 双光子显微镜是一种对细胞和细胞外基质结构进行亚微米分辨率活体成像的有力工具。基于双光子的非侵入性光学活检方法具有用作切除活检和组织病理学的辅助手段的潜力。虽然双光子显微镜在生物学研究中的实用性已经在 在神经生物学和胚胎学等领域，其临床潜力仍未实现。操作复杂性、尺寸、成像速度和成本阻碍了在临床环境中测试这项新技术。该提议通过设计紧凑的双光子光学活检探针来解决这些困难。将构建手持设备，以视频速率（> 10帧/秒）对组织细胞结构进行成像，亚细胞分辨率低至150-200 - 1的深度。已经完成了一些初步研究，以证明这一项目的可行性。(1)双光子激发在组织细胞成像中的应用 的结构和代谢的基础上，其自身荧光被证明，初步数据表明，这种技术的潜力，以区分正常和恶性组织包括在内。(2)一个原型的视频率双光子显微镜的构建。(3)一个共焦反射光成像子系统被纳入一个双光子显微镜。(4)基于双光子光谱，可以分辨出组织生化成分的分布。 (5)本研究的目的是开发一种适合于临床研究的手持式双光子活检探针。我们将研究与双光子技术相关的工程挑战，如超短光脉冲的传输。避免组织光损伤也是至关重要的; 将建立适当的激光功率和剂量水平。我们将在组织模型、动物模型和切除的人体皮肤活检标本中表征该器械的性能。我们希望，该项目的成功完成将导致第一代设备，将允许在临床双光子光学活检技术的评估。

DESCRIPTION (provided by applicant): Two-photon microscope is a powerful tool for in vivo imaging of cellular and extracellular matrix structures with sub-micron resolution. Two-photon based non-invasive optical biopsy methods have the potential of being used as an adjunct to excisional biopsy and histopathology. While the utility of two-photon microscopy for biological studies has been clearly demonstrated in areas such as neurobiology and embryology, its clinical potential remains unrealized. The operational complexity, size, imaging speed and cost stand in the way of testing this new technology in a clinical setting. This proposal addresses these difficulties by engineering a compact two-photon optical biopsy probe. A hand-held device will be built to image tissue cellular structures at video rate (> 10 frames/sec) with sub-cellular resolution down to a depth of 150-200 -1. A number of preliminary studies have been completed to demonstrate the feasibility of this project. (1) The use of two- photon excitation to image tissue cellular structures and metabolism based on its autofluorescence was demonstrated, Preliminary data indicating this technique's potential to distinguish normal and malignant tissues are included. (2) A prototype video-rate two-photon microscope was constructed. (3) A confocal reflected-light imaging sub-system was incorporated into a two-photon microscope. (4) The distribution of tissue biochemical constituents has been resolved based on their two-photon spectra. (5) The key two-photon photodamage mechanisms were identified, The aim of this proposal is to develop a hand-held two-photon biopsy probe suitable for clinical research. We will study the engineering challenges associated with miniaturizing two-photon technology such as the delivep ultra-short light pulses. It is also critical to avoid tissue photodamage; the maximum perr&sibl.e laser power and dosage levels will be established. We will characterize the performance this device in tissue phantoms, animal models and excised human skin biopsy specimens. We hope that the successful completion of this project will result in a first generation device that will allow the evaluation of two-photon optical biopsy techniques in the clinics.