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.

概括 对哺乳动物免疫小球的研究及其与组织的相互作用对于理解至关重要 它们如何调节从伤口愈合到自身免疫性疾病开始到癌症的过程 设计更好的治疗策略来治疗这些普遍的疾病。插入性多光子显微镜 （MPM）与荧光记录器小鼠模型以及体内细胞和组织标记的丰富曲目结合 技术使在皮肤亚细胞水平和 其他器官。但是，人类皮肤的结构和免疫环境存在显着差异 将这些发现的转换性限制为人皮肤免疫反应。我们的小组最近有 开发了一个快速大面积的多光外观（火焰），这是一个优化的独特成像平台，可高效 临床皮肤成像以快速产生具有微观分辨率的宏观图像（MM至CM尺度）（0.5-- 1µm）基于无标签的分子对比度（荧光强度和寿命）。在此应用程序中，我们利用 我们在MPM技术开发和400多名患者的临床成像方面的丰富经验 过去几年，开发第一个基于MPM的临床设备（IFLAME）作为研究成像工具 优化并专门用于免疫群体种群的无体体标签成像及其动力学 人皮。在AIM 1中，我们开发了IFLAME作为一种临床研究工具，用于有效的体内标签成像 真皮细胞群体及其在人体皮肤中的动态。这项工作涉及开发检测和 分析方法以及光学和计算方法，以实现快速荧光寿命检测 和分析以自动化细胞形态和代谢特征的测量。在 AIM 2，我们通过在正常中证明免疫球的体内表征来验证IFLAME性能 并发炎人类皮肤。在AIM 3中，我们开发定量形态和代谢MPM成像 在监测伤口的背景下，评估免疫浸润物及其动态的终点 康复。这项工作代表了使用MPM与图像，识别和形成对比的MPM固有源的首次尝试 根据其光学特征和迁移行为来量化人体皮肤中的关键免疫细胞。我们的 长期目标是开发IFLAME作为一种临床研究工具，用于快速，无标签的免疫核管成像 基于细胞形态和代谢成像终点的皮肤。这些可用于更好地理解 评估并优化伤口愈合，自身免疫性皮肤疾病和治疗反应。

项目成果

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

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

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.