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

基本信息

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
项目类别：
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

项目摘要

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免疫细胞至关重要它们如何调节从伤口愈合到自身免疫性疾病发生再到癌症的过程设计更好的治疗策略来治疗这些常见疾病。 (MPM) 结合丰富的荧光报告小鼠模型和体内细胞和组织标记技术使得在皮肤和细胞的亚细胞水平上可视化免疫细胞与组织的相互作用成为可能。然而，人类皮肤的结构和免疫环境存在显着差异。限制了这些发现对人类皮肤免疫反应的可转化性。开发了快速大面积多光子外窥镜（FLAME），这是一种针对高效进行优化的独特成像平台临床皮肤成像可快速生成具有微观分辨率（0.5- 1μm）基于无标记分子对比（荧光强度和寿命）。我们在 MPM 技术开发和超过 400 名患者的临床成像方面拥有丰富的经验过去几年开发出第一个基于 MPM 的临床设备 (iFLAME) 作为研究成像工具针对免疫细胞群及其动态的体内无标记成像进行了优化并致力于在目标 1 中，我们开发了 iFLAME 作为临床研究工具，用于高效的体内无标记成像。这项工作涉及检测和研究人类皮肤中的真皮细胞群及其动态。分析方法以及光学和计算方法可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需的分析。目标 2，我们通过展示正常情况下免疫细胞的体内特征来验证 iFLAME 性能在目标 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的免疫浸润及其动态的终点这项工作代表了首次尝试使用 MPM 对比的内在来源来进行图像、识别和治疗。根据体内人体皮肤中的关键免疫细胞的光学特征和迁移行为对其进行量化。长期目标是将 iFLAME 开发为一种临床研究工具，用于对免疫细胞进行快速、无标记成像基于细胞形态和代谢成像终点的皮肤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

期刊论文数量（5）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

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

使用快速大面积多光子外窥镜（FLAME）进行体内成像，捕捉人体皮肤中黑色素分布的不均匀性。

DOI：
发表时间：
2022-05-16
期刊：
影响因子：
4.6
作者：
Vicente, Juvinch R;Durkin, Amanda;Shrestha, Kristina;Balu, Mihaela
通讯作者：
Balu, Mihaela

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：
发表时间：
2023-08-17
期刊：
影响因子：
0
作者：
Shiu, Jessica;Lentsch, Griffin;Polleys, Christopher M;Mobasher, Pezhman;Ericson, Marissa;Georgakoudi, Irene;Ganesan, Anand K;Balu, Mihaela
通讯作者：
Balu, Mihaela

Vitreoretinal Surgical Instrument Tracking in Three Dimensions Using Deep Learning.

使用深度学习进行三维玻璃体视网膜手术器械跟踪。

DOI：
发表时间：
2023-01-03
期刊：
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

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

深度学习辅助多光子显微镜可减少光暴露并加快高光敏感度和低光敏感度组织的成像。

DOI：
发表时间：
2021
期刊：
影响因子：
3
作者：
McAleer, Stephen;Fast, Alexander;Xue, Yuntian;Seiler, Magdalene J;Tang, William C;Balu, Mihaela;Baldi, Pierre;Browne, Andrew W
通讯作者：
Browne, Andrew W

In vivo multiphoton microscopy of melasma.

黄褐斑的体内多光子显微镜。

DOI：
发表时间：
2019
期刊：
影响因子：
4.3
作者：
Lentsch, Griffin;Balu, Mihaela;Williams, Joshua;Lee, Sanghoon;Harris, Ronald M;König, Karsten;Ganesan, Anand;Tromberg, Bruce J;Nair, Nirmala;Santhanam, Uma;Misra, Manoj
通讯作者：
Misra, Manoj