Two-Photo Optical Biopsy Probe

描述(由申请人提供)： 双光子显微镜是在体亚微米分辨率的细胞和细胞外基质结构成像的有力工具。基于双光子的非侵入性光学活检方法有可能被用作切除活检和组织病理学的辅助手段。虽然双光子显微镜在生物学研究中的应用已经在 在神经生物学和胚胎学等领域，其临床潜力仍未实现。操作的复杂性、大小、成像速度和成本阻碍了在临床环境中测试这项新技术。该方案通过设计一种紧凑的双光子光学活组织检查探头来解决这些困难。将建造一个手持设备，以视频速率(&gt；10帧/秒)成像组织细胞结构，亚细胞分辨率降至150-200-1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)利用双光子激发对组织细胞成像 展示了基于其自身荧光的结构和代谢，包括表明该技术在区分正常组织和恶性组织方面的潜力的初步数据。(2)搭建了视频率双光子显微镜样机。(3)在双光子显微镜中加入共聚焦反射光成像子系统。(4)根据组织生化成分的双光子光谱，对其分布进行了解析。 (5)确定了双光子光损伤的关键机制，旨在研制一种适合于临床研究的手持式双光子活检探头。我们将研究与小型化双光子技术相关的工程挑战，例如Delivep超短光脉冲。避免组织光损伤也是至关重要的；最大 Perr&Sibl.e激光功率和剂量水平将确定。我们将在组织模体、动物模型和切除的人类皮肤活检标本中表征该设备的性能。我们希望这个项目的成功完成将产生第一代设备，使双光子光学活组织检查技术能够在临床上进行评估。

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.