3D Fourier Imaging System for High Throughput Analyses of Cancer Organoids

用于癌症类器官高通量分析的 3D 傅里叶成像系统

• 批准号: 10577796
• 负责人: Hakho Lee
• 金额: $ 19.24万
• 依托单位: MASSACHUSETTS GENERAL HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-03-01 至 2025-02-28
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10577796
• 关键词: 3-Dimensional; Accounting; Adopted; Algorithms; Antineoplastic Agents; Antitumor Drug Screening Assays; Apoptotic; Beds; Brightfield Microscopy; Cells; Cellularity; Classification; Clinical; Code; Color; Computer software; Data; Data Set; Detection; Drug resistance; Drug usage; Environment; Epithelium; Extracellular Matrix; Goals; Heterogeneity; Image; Individual; Infusion procedures; Lateral; Light; Lighting; Machine Learning; Malignant Neoplasms; Mechanics; Mesenchymal; Methods; Microfluidics; Microscope; Microscopy; Modeling; Molecular; Monitor; Optics; Organoids; Outcome; Patients; Pharmaceutical Preparations; Pharmacotherapy; Phenotype; Physiological; Positioning Attribute; Proliferating; Radiation; Research Personnel; Resistance; Resolution; Scanning; Source; Specimen; Speed; Stimulus; Structure; System; Systems Development; Techniques; Testing; Training; Treatment outcome; Visual; advanced analytics; anticancer research; cancer imaging; cellular imaging; chemotherapy; computational pipelines; computerized data processing; data acquisition; deep learning; deep learning algorithm; design; drug development; drug resistance development; empowerment; experimental study; feature extraction; high resolution imaging; high throughput analysis; imaging platform; imaging system; improved; in vivo; innovation; meter; microscopic imaging; multidimensional data; predicting response; prospective; reconstruction; resistance mechanism; response; self organization; single cell analysis; statistics; temozolomide; three dimensional cell culture; tomography; tool; trait; tumor; tumor heterogeneity

Challenges. Tumor spheroids (and organoids) have become an instrumental tool in cancer research. These self-organized, three-dimensional (3D) systems can recapitulate phenotypic and functional traits of patient tumors in vivo, thereby serving as a powerful testing bed to study tumor heterogeneity, interactions with the environment (e.g., extracellular matrix), and responses to external stimuli (e.g., chemotherapy, radiation). Fully harnessing spheroids' utility, however, is stymied by lack of high-throughput analysis methods. Conventional bright-field microscopy, although widely used to monitor spheroids in culture, fails to capture detailed cellular organizations; advanced fluorescent microscopy can resolve individual cells, but its imaging throughput is restricted by the small field-of-view (FOV) and the scanning mechanisms involved. Innovations. We aim to advance a new volumetric imaging microscope (VIM) for single cell analyses in tumor spheroids. Specifically, we will explore integrating Fourier ptychographic microscopy (FPM) with diffraction tomography. FPM is based on a spatially coded-illumination technique, collecting low resolution image sequences while changing the position of a point-light source. These images are then numerically combined in the Fourier space, which allows FPM to achieve both wide field-of-view and high spatial resolution in 2D images. We reason that full 3D microscopic images can be recovered by accounting for optical diffraction during the numerical reconstruction. Approaches. Aim 1. System development. We will build a VIM system featuring: i) a new numerical algorithm to reconstruct 3D volumetric images; ii) a new light-illumination strategy to speed up the data acquisition; iii) microfluidic cartridges optimized for spheroid culture and drug treatment; and iv) multicolor imaging capacity for molecular detection. The complete VIM will resolve individual cells constituting a spheroid at high resolution (lateral, 0.4 µm; axial, 1 µm) in a large imaging volume. Aim 2. Treatment monitoring with tumor spheroids. We will test VIM's practical utility: VIM-enabled spheroid imaging will reveal earlier than bulk imaging whether a spheroid is responsive or resistance to drug treatment. To generate a tumor model, we will use primary GBM cells from patients. GBM spheroids will be grown and treated with drug (temozolomide) inside microfluidic cartridges. We will use the VIM to monitor how single cells change their phenotypes under treatment, and correlate these changes with treatment outcomes. Impact. The VIM will be a transformative tool for cancer research, empowering researchers with rich data sets and substantially advanced analytics. Immediate applications include better monitoring of anticancer drug responses in 3D cell culture, analyzing cellular heterogeneity, and prospectively detecting cellular fate under various physiological conditions. These outcomes will strengthen the clinical and scientific utility of tumor spheroids in cancer research.

挑战：肿瘤球体（和类器官）已成为癌症研究的重要工具。 自组织的三维 (3D) 系统可以概括患者的表型和功能特征 体内肿瘤，从而作为研究肿瘤异质性、与肿瘤的相互作用的强大测试平台 环境（例如细胞外基质）和对外部刺激（例如化疗、放疗）的反应。 然而，由于缺乏传统的高通量分析方法，利用球体的实用性受到阻碍。 明场显微镜虽然广泛用于监测培养中的球体，但无法捕获详细的细胞 组织；先进的荧光显微镜可以解析单个细胞，但其成像吞吐量 受到小视场（FOV）和所涉及的扫描机制的限制。 开发一种新型体积成像显微镜 (VIM)，用于肿瘤球体中的单细胞分析。 我们将探索将傅里叶叠层成像 (FPM) 与衍射断层扫描相结合。 基于空间编码照明技术，收集低分辨率图像序列，同时改变 然后将这些图像在傅里叶空间中进行数字组合，从而得到点光源的位置。 允许 FPM 在 2D 图像中实现宽视场和高空间分辨率，我们推断全 3D 图像。 可以通过在数值重建过程中考虑光学衍射来恢复显微图像。 目标 1. 系统开发 我们将构建一个具有以下特点的 VIM 系统： i) 一种新的数值算法。 重建 3D 体积图像；ii) 加速数据采集的新光照策略； 针对球体培养和药物治疗进行了优化的微流体盒；以及 iv) 多色成像能力； 完整的 VIM 将以高分辨率解析构成球体的单个细胞。 （横向，0.4 µm；轴向，1 µm）在大成像体积中。 目标 2. 使用肿瘤球体进行治疗监测。 将测试 VIM 的实用性：支持 VIM 的球体成像将比批量成像更早地揭示是否 为了生成肿瘤模型，我们将使用原发性 GBM。 来自患者的 GBM 球体将在微流体内生长并用药物（替莫唑胺）进行处理。 我们将使用 VIM 监测单细胞在治疗过程中如何改变其表型，以及 VIM 将成为癌症治疗的变革性工具。 研究，为研究人员提供丰富的数据集和即时先进的分析。 应用包括更好地监测 3D 细胞培养中的抗癌药物反应、分析细胞 异质性，并前瞻性地检测各种生理条件下的细胞命运。 结果将加强肿瘤球体在癌症研究中的临床和科学实用性。