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: 9904165
• 负责人: Mihaela Balu
• 金额: $ 48.32万
• 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 2022-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9904165
• 关键词: Address; Adenosine; Adoption; Affect; Basal cell carcinoma; Benchmarking; Biology; Cells; Cellular Morphology; Characteristics; Clinical; Clinical Research; Collagen; Computers; Contrast Sensitivity; Cutaneous; DNA Damage; Data; Detection; Development; Devices; Diagnosis; Dinucleoside Phosphates; Discrimination; Dose; Elastin Fiber; Electronics; Environment; Epithelial; Epithelium; Evaluation; Fiber; Flavins; Fluorescence; Generations; Germany; Goals; Head; Heterogeneity; Image; Keratin; Label; Lasers; Light; Low-Level Laser Therapy; Maps; Measures; Mechanics; Melanins; Metabolic; Metabolism; Microscope; Mitochondria; Modality; Molecular; Morphology; Nicotinamide adenine dinucleotide; Noise; Optics; Organ Transplantation; Patient imaging; Patients; Performance; Phenotype; Photons; Pigments; Pilot Projects; Population; Positioning Attribute; Procedures; Process; Resolution; Safety; Scanning; Signal Transduction; Skin; Skin Pigmentation; Skin Tissue; Skin graft; Source; Speed; Structure; System; Technology; Therapeutic; Tissues; Translations; Transplantation; Vitiligo; arm; base; cell injury; cellular imaging; clinical imaging; contrast enhanced; cost; cost effective; design; epithelial wound; experience; healing; healthy volunteer; improved; in vivo; innovation; instrument; keratinocyte; lens; melanocyte; melanoma; metabolic imaging; microscopic imaging; migration; molecular imaging; multi-photon; multiphoton microscopy; next generation; portability; prototype; quantitative imaging; response; safety assessment; second harmonic; skin disorder; standard of care; submicron; success; technology development; tool; translational study; treatment response; two-photon; wound; wound healing

PROJECT SUMMARY Multiphoton microscopy (MPM) can provide sub-micron resolution images of living tissues in their native environment with label-free molecular contrast from multiple modalities, including second harmonic generation (SHG) and two-photon excited fluorescence (TPEF). Several endogenous tissue components can be visualized, including collagen (from SHG) and reduced nicotinamide adenine dinucleotide (NADH), flavin adenosine dinucleotide (FAD), keratin, melanin and elastin fibers (from TPEF). We have advanced label-free MPM technologies in skin clinical/translational studies for characterizing keratinocyte metabolism, diagnosing melanoma, understanding melanocyte biology, detecting basal cell carcinoma, quantifying skin pigmentation, and assessing the effects of cutaneous laser therapy. Many of these studies have been completed using a commercial multi-photon microscope for clinical skin imaging that has limitations in terms of field-of-view (FOV), speed, footprint, and cost. In order to address these barriers to clinical adoption, we propose to build a “next-generation” clinical multiphoton microscope that integrates advanced benchtop technologies into a compact, practical, and cost-effective bedside device. This new instrument will have comparable FOV, resolution, and scanning features to standard-of-care reflectance confocal microscopes (RCM), yet provide unique structural and metabolic contrast from multiple modalities (TPEF and SHG) that can only be achieved with MPM. We will establish the clinical safety of this device in a light dose escalation study that assesses DNA and cellular damage, and establish key performance benchmarks in a 12-patient clinical study of healthy volunteers across a range of skin types. In addition, we will conduct pilot studies of wound re-epithelialization and melanocyte migration in the context of vitiligo micro-grafts, a clinical procedure where pigmented skin is transplanted into skin affected by vitiligo, which is devoid of melanocytes. Melanocytes migrating out of engrafted skin and keratinocytes turning over within engrafted skin can be visualized by measuring the TPEF of cellular melanin and co-factors (NADH, FAD+) in and around the grafts, effectively identifying different cell populations involved in wound healing. Our broad, long term goal is to develop ev-MPM as a practical approach for rapid, in vivo characterization of cellular morphologic and metabolic imaging endpoints in patients. These can be used to understand and optimize wound healing and provide a practical beside platform for detecting, diagnosing, and optimizing therapeutic response in skin diseases.

项目概要 多光子显微镜（MPM）可以提供活体组织的亚微米分辨率图像 具有来自多种模式（包括二次谐波生成）的无标记分子对比的环境 （SHG）和双光子激发荧光（TPEF）可以是几种内源性组织成分。 可视化，包括胶原蛋白（来自 SHG）和还原型烟酰胺腺嘌呤二核苷酸 (NADH)、黄素 腺苷二核苷酸 (FAD)、角蛋白、黑色素和弹性蛋白纤维（来自 TPEF）。 MPM 技术在皮肤临床/转化研究中用于表征角质形成细胞代谢、诊断 黑色素瘤、了解黑色素细胞生物学、检测基底细胞癌、量化皮肤色素沉着、 并评估皮肤激光治疗的效果，其中许多研究已经使用a完成。 用于临床皮肤成像的商用多光子显微镜在视野方面存在局限性 （FOV）、速度、占地面积和成本为了解决临床采用的这些障碍，我们建议建立。 “下一代”临床多光子显微镜，将先进的台式技术集成到 紧凑、实用且具有成本效益的床边设备将具有可比的 FOV、 分辨率和扫描功能达到标准护理反射共焦显微镜 (RCM) 的水平，但还提供 多种模式（TPEF 和 SHG）的独特结构和代谢对比只能实现 我们将通过 MPM 进行轻剂量递增研究来确定该设备的临床安全性。 DNA 和细胞损伤，并在 12 名健康患者的临床研究中建立关键性能基准 此外，我们还将针对伤口上皮再生进行试点研究。 和白癜风微移植背景下的黑素细胞迁移，这是一种将色素皮肤移植到皮肤上的临床手术 移植到受白癜风影响的皮肤中，该皮肤缺乏黑色素细胞迁移出来。 可以通过测量 TPEF 来观察移植皮肤和移植皮肤内角质形成细胞的变化 移植物内部和周围的细胞黑色素和辅助因子（NADH、FAD+），有效识别不同的细胞 我们广泛的长期目标是将 ev-MPM 开发为一种实用的方法。 快速体内表征细胞形态和代谢成像终点的方法 这些可用于了解和优化伤口愈合，并提供一个实用的旁平台。 用于检测、诊断和优化皮肤病的治疗反应。