Development of a fast scanning, extended field-of-view multiphoton microscope for clinical skin imaging

开发用于临床皮肤成像的快速扫描、扩展视场多光子显微镜

• 批准号: 10680741
• 负责人: Mihaela Balu
• 金额: $ 58.04万
• 依托单位: UNIVERSITY OF CALIFORNIA-IRVINE
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-07-01 至 2027-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10680741
• 关键词: Affect; Algorithms; Area; Atopic Dermatitis; Autoimmune; Autoimmune Diseases; Behavior; Benchmarking; Biological Markers; Cells; Cellular Assay; Cellular Morphology; Chemicals; Clinical; Clinical Research; Collagen; Compensation; Computing Methodologies; Cutaneous; Data; Dedications; Dermal; Dermis; Detection; Development; Devices; Elastin; Epithelium; Fiber; Fluorescence; Funding; Generations; Genetic Transcription; Goals; Grant; Histologic; Human; Image; Imaging Device; Immune; Immune response; Immunologic Monitoring; In Situ; Infiltration; Keratin; Label; Lasers; Location; Macrophage; Malignant Neoplasms; Measurement; Melanins; Metabolic; Methods; Microscope; Microscopic; Modality; Molecular; Morphology; Movement; NADH; Optical Methods; Optics; Organ; Organ Transplantation; Patients; Performance; Physiologic pulse; Play; Population; Process; Reporter; Research; Resolution; Scanning; Signal Transduction; Skin; Source; Specificity; Speed; Structure; Techniques; Technology; Therapeutic; Time; Tissues; Visualization; Work; cell motility; cellular imaging; clinical imaging; cofactor; contrast imaging; convolutional neural network; data acquisition; design; detection sensitivity; experience; fluorophore; healing; human imaging; imaging platform; immune cell infiltrate; immune imaging; improved; in vivo; instrument; intravital imaging; metabolic imaging; microscopic imaging; mouse model; multi-photon; multiphoton microscopy; operation; portability; protein biomarkers; prototype; second harmonic; skin disorder; standard of care; technology development; tool; treatment response; tumor-immune system interactions; two-photon; wound; wound care; wound healing; wound treatment

Summary The study of mammalian immune cells and their interactions with tissue in situ is critical for understanding how they regulate processes ranging from wound healing to autoimmune disease initiation to cancer and for designing better therapeutic strategies to treat these prevalent conditions. Intravital multiphoton microscopy (MPM) combined with a rich repertoire of fluorescent reporter mouse models and in vivo cell and tissue labeling techniques have made it possible to visualize immune cell-tissue interactions at a subcellular level in skin and other organs. However, there are significant differences in the structure and immune milieu of human skin that limits the translatability of these findings to the human cutaneous immune response. Our group has recently developed a fast large area multiphoton exoscope (FLAME), a unique imaging platform optimized for efficient clinical skin imaging to rapidly generate macroscopic images (mm to cm-scale) with microscopic resolution (0.5- 1µm) based on label-free molecular contrast (fluorescence intensity and lifetime). In this application, we leverage our extensive experience in MPM technology development and clinical imaging of more than 400 patients over the past several years to develop the first MPM-based clinical device (iFLAME) as a research imaging tool optimized for, and dedicated to, in vivo label-free imaging of immune cell populations and their dynamics in human skin. In Aim 1, we develop iFLAME as a clinical research tool for efficient in vivo label-free imaging of dermal cell populations and their dynamics in human skin. This work involves development of detection and analytic approaches as well as optical and computational methods to enable rapid fluorescence lifetime detection and analysis necessary to automate measurements of the cellular morphological and metabolic signatures. In Aim 2, we validate iFLAME performance by demonstrating in vivo characterization of immune cells in normal and inflamed human skin. In Aim 3, we develop quantitative morphological and metabolic MPM imaging endpoints to assess immune infiltrates and their dynamics in human skin in the context of monitoring wound healing. This work represents the first attempt to use intrinsic sources of MPM contrast to image, identify, and quantify key immune cells in human skin in vivo based on their optical signatures and migratory behavior. Our long-term goal is to develop iFLAME as a clinical research tool for rapid, label-free imaging of immune cells in skin based on cellular morphologic and metabolic imaging endpoints. These can be used to better understand, evaluate and optimize wound healing, autoimmune skin diseases and therapeutic responses.

总结 研究哺乳动物免疫细胞及其与原位组织的相互作用对于理解 它们是如何调节从伤口愈合到自身免疫性疾病引发到癌症以及 设计更好的治疗策略来治疗这些普遍的疾病。活体多光子显微术 (MPM)结合丰富的荧光报告小鼠模型库和体内细胞和组织标记 技术使得在皮肤中亚细胞水平上观察免疫细胞-组织相互作用成为可能， 其他器官。然而，人体皮肤的结构和免疫环境存在显著差异， 限制了这些发现对人类皮肤免疫应答的可转化性。我们集团最近 开发了一种快速大面积多光子外窥镜（FLAME），这是一种独特的成像平台， 临床皮肤成像，以快速生成具有显微分辨率（0.5- 2.5 μ m）的宏观图像（mm至cm尺度）， 1µm），基于无标记分子对比度（荧光强度和寿命）。在这个应用程序中，我们利用 我们在MPM技术开发和超过400例患者的临床成像方面的丰富经验 在过去的几年中，开发了第一个基于MPM的临床设备（iFLAME）作为研究成像工具 优化并致力于免疫细胞群体的体内无标记成像及其在体内的动力学， 人类皮肤在目标1中，我们开发了iFLAME作为一种临床研究工具，用于有效的体内无标记成像， 真皮细胞群及其在人体皮肤中的动态。这项工作涉及发展检测和 分析方法以及光学和计算方法，以实现快速荧光寿命检测 以及自动测量细胞形态和代谢特征所必需的分析。在 目的2，我们通过证明正常人中免疫细胞的体内表征来验证iFLAME性能。 和发炎的人类皮肤在目标3中，我们开发了定量形态学和代谢MPM成像， 在监测伤口的背景下评估人体皮肤中免疫浸润及其动力学的终点 治愈这项工作代表了第一次尝试使用MPM对比度的内在来源来成像，识别， 基于它们的光学特征和迁移行为在体内量化人类皮肤中的关键免疫细胞。我们 长期目标是开发iFLAME作为临床研究工具，用于快速，无标记的免疫细胞成像， 基于细胞形态学和代谢成像终点的皮肤。这些可以用来更好地理解， 评估和优化伤口愈合、自身免疫性皮肤病和治疗反应。

项目成果

Deep Learning-Assisted Multiphoton Microscopy to Reduce Light Exposure and Expedite Imaging in Tissues With High and Low Light Sensitivity.

• DOI: 10.1167/tvst.10.12.30
• 发表时间: 2021-10-04
• 期刊: Translational vision science & technology
• 影响因子: 3
• 作者: McAleer S;Fast A;Xue Y;Seiler MJ;Tang WC;Balu M;Baldi P;Browne AW
• 通讯作者: Browne AW

Non-invasive Imaging Techniques for Monitoring Cellular Response to Treatment in Stable Vitiligo.

用于监测细胞对稳定白癜风治疗反应的非侵入性成像技术。

• DOI: 10.1101/2023.08.15.553419
• 发表时间: 2023
• 期刊: bioRxiv : the preprint server for biology
• 作者: Shiu,Jessica;Lentsch,Griffin;Polleys,ChristopherM;Mobasher,Pezhman;Ericson,Marissa;Georgakoudi,Irene;Ganesan,AnandK;Balu,Mihaela
• 通讯作者: Balu,Mihaela

In vivo imaging with a fast large-area multiphoton exoscope (FLAME) captures the melanin distribution heterogeneity in human skin.

• DOI: 10.1038/s41598-022-12317-y
• 发表时间: 2022-05-16
• 期刊: Scientific reports
• 影响因子: 4.6

Vitreoretinal Surgical Instrument Tracking in Three Dimensions Using Deep Learning.

• DOI: 10.1167/tvst.12.1.20
• 发表时间: 2023-01-03
• 期刊: TRANSLATIONAL VISION SCIENCE & TECHNOLOGY
• 影响因子: 3
• 作者: Baldi, Pierre F.;Abdelkarim, Sherif;Liu, Junze;To, Josiah K.;Ibarra, Marialejandra Diaz;Browne, Andrew W.
• 通讯作者: Browne, Andrew W.