Optical Amplification Microscopy of Weak Back-Scattered Light

弱背散射光的光学放大显微镜

• 批准号: 9214154
• 负责人: Stephen A Boppart
• 金额: $ 54.71万
• 依托单位: UNIVERSITY OF ILLINOIS AT URBANA-CHAMPAIGN
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-09-01 至 2020-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9214154
• 关键词: Address; Amplifiers; Apoptosis; Back; Ballistics; Basic Science; Biological; Biological Process; Cell Culture Techniques; Cells; Clinical Research; Complex; Confocal Microscopy; Data; Detection; Development; Diagnostic; Electronics; Environment; Event; Feasibility Studies; Fluorescence; Future; Generations; Goals; Image; Imaging technology; In Vitro; Label; Life; Light; Medical; Microscope; Microscopy; Modality; Molecular; Multimodal Imaging; Mus; Noise; Optical Coherence Tomography; Optics; Performance; Photons; Physics; Process; Research; Resolution; Sampling; Sensitivity and Specificity; Signal Transduction; Skin; Source; Speed; Techniques; Technology; Tissues; base; bioimaging; cellular imaging; clinical Diagnosis; clinical application; clinical investigation; clinical practice; contrast imaging; cost; detector; fascinate; imaging modality; imaging system; improved; in vivo; in vivo imaging; insight; instrument; instrumentation; intravital imaging; light scattering; microscopic imaging; multi-photon; novel; optical imaging; practical application; pre-clinical; second harmonic; temporal measurement; two-photon

SUMMARY Optical imaging, which offers sufficient spatial resolution, specificity, sensitivity, and temporal resolution, has provided substantial insights into important biological processes at the cellular and molecular levels. Although high quality optical images can be obtained from in vitro cell cultures and/or thin tissue sections, intravital cell imaging in a complex, three-dimensional living tissue environment remains quite challenging. Because biological tissue naturally does not favor the propagation of light, and because of the inevitable presence of strong ambient light in the environment, special challenges arise in virtually every in vivo biomedical optical imaging, namely weak light signals and strong light backgrounds (including the multiply-scattered light in the tissue and the environment light). These challenges have significantly limited the full application of optical imaging in in vivo pre-clinical and clinical investigations. Currently, the detection of weak light signals for optical imaging relies heavily on the use of highly sensitive electronic detectors and electronic amplifiers. Although high-end electronic photo receivers are often very sensitive, the high sensitivity leads to the imaging systems being extremely prone to random photon noise, such environmental background (room) light, which is problematic for practical applications (e.g. in vivo studies and clinical practice). Furthermore, electronic detectors are incapable of distinguishing image-bearing ballistic photons from the multiply-scattered light background, which as a predominant source of noise in optical imaging of biological samples, can be overwhelming and significantly degrade resolution when imaging microstructure deep in tissue. In this proposed project, we will develop a multimodal microscope that utilizes a novel high speed optical parametric amplifier (OPA) to optically amplify weak back-scattered light signals, and demonstrate its capabilities by investigating in vivo cellular apoptosis events in murine skin. As shown by our preliminary data, the OPA will not only provide a high level of signal gain to improve detection sensitivity, but also provide an inherent nonlinear optical gate to both extract imaging-bearing signals and reject the noise sources from environmental photons and multiply-scattered background light. We will systematically explore the benefits afforded by this OPA for multimodal imaging that will include label-free reflectance confocal microscopy and optical coherence tomography. Improvement in resolution, contrast, imaging depth, and reduced photo-damage, will be investigated. The successful completion of this project will demonstrate a high-speed, robust, optical intravital microscope that combines multiple modalities with enhanced performance and new fascinating imaging functions uniquely enabled by the OPA. This intravital microscope will not only enable new biological and clinical studies, but also promote the development of new optical imaging technologies based on optical amplifiers.

概括 光学成像提供足够的空间分辨率，特异性，灵敏度和时间分辨率，具有 对细胞和分子水平的重要生物过程提供了大量见解。虽然 可以从体外细胞培养物和/或薄组织，插入式细胞中获得高质量的光学图像 在复杂的三维生活组织环境中进行成像仍然很具有挑战性。因为生物学 组织自然不利于光的传播，并且由于不可避免地存在强烈的环境 环境中的光线，几乎每个体内生物医学光学成像都会出现特殊挑战，即 弱的光信号和强光背景（包括组织中的多重散射光和 环境光）。这些挑战极大地限制了光学成像在体内的全面应用 临床前和临床研究。目前，检测光学成像的弱光信号依赖 大量使用高度敏感的电子探测器和电子放大器。虽然高端电子 照片接收器通常非常敏感，高灵敏度会导致成像系统非常容易发生 对于随机的光子噪声，这种环境背景（房间）光，这对于实用性是有问题的 应用（例如体内研究和临床实践）。此外，电子探测器无法 将带有图像的弹道光子与多散射光背景区分开来 在生物样品的光学成像中，主要的噪声来源可能是压倒性的，显着的 在组织深处成像微观结构时，降解分辨率。 在这个拟议的项目中，我们将开发一个多模式显微镜，该显微镜利用了新型的高速光学 参数放大器（OPA）以光学地放大弱的背面光信号，并演示其功能 通过研究鼠皮肤中体内细胞凋亡事件。如我们的初步数据所示，OPA将 不仅提供高水平的信号增益以提高检测灵敏度，而且还提供了固有的非线性 既要提取成像信号的光学门，又拒绝来自环境光子的噪声源 并乘以散落的背景光。我们将系统地探索此OPA所提供的好处 多模式成像，其中包括无标签反射率共焦显微镜和光学连贯性 断层扫描。分辨率，对比度，成像深度和减少的照片损伤的改善将是 调查。该项目的成功完成将展示高速，健壮，光插入 显微镜将多种方式与增强性能和新的迷人成像功能相结合 OPA独特地启用。这种插入显微镜不仅可以实现新的生物学和临床研究，还可以 但也促进基于光学放大器的新光学成像技术的开发。