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: 10358186
• 负责人: Hakho Lee
• 金额: $ 19.64万
• 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/10358186
• 关键词: 3-Dimensional; Accounting; Adopted; Algorithms; Antineoplastic Agents; Antitumor Drug Screening Assays; Apoptotic; Beds; Brightfield Microscopy; Cell Culture Techniques; Cells; Cellularity; Chemotherapy and/or radiation; Clinical; Code; Computer software; Data; Data Set; Detection; Drug resistance; Drug usage; Environment; Epithelial; Extracellular Matrix; Goals; Gold; 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; Research Personnel; Resistance; Resolution; Scanning; Source; Specimen; Speed; Stimulus; Structure; Supervision; System; Systems Development; Techniques; Testing; Training; Treatment outcome; Visual; advanced analytics; anticancer research; base; cancer imaging; cellular imaging; computational pipelines; computerized data processing; data acquisition; deep learning; deep learning algorithm; design; empowered; experimental study; feature extraction; high resolution imaging; high throughput analysis; imaging platform; imaging system; in vivo; innovation; microscopic imaging; multidimensional data; predicting response; prospective; reconstruction; resistance mechanism; response; single cell analysis; statistics; temozolomide; 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（和器官）已成为癌症研究中的工具工具。这些 自组织的三维（3D）系统可以概括患者的表型和功能性状 体内肿瘤，从而充当强大的测试床以研究肿瘤异质性，与 环境（例如细胞外基质）和对外部刺激的反应（例如化学疗法，放射线）。完全 然而，利用球体的效用受到缺乏高通量分析方法的阻碍。传统的 明亮的轨道显微镜虽然广泛用于监测培养的球形素，但未能捕获详细的细胞 组织；高级荧光显微镜可以解析单个细胞，但其成像吞吐量为 受小型视图（FOV）和所涉及的扫描机制的限制。创新。我们的目标 推进新的体积成像显微镜（VIM），以用于肿瘤球形的单细胞分析。特定的 我们将探索与衍射断层扫描相结合的傅立叶ptychographic显微镜（FPM）。 FPM是基于的 在空间编码的弹片技术上，收集低分辨率图像序列，同时更改 点光源的位置。然后将这些图像在数值中组合在傅立叶空间中， 允许FPM在2D图像中同时获得宽阔的视图和高空间分辨率。我们认为完整的3D 可以通过考虑数值重建过程中的光学衍射来恢复微观图像。 方法。目标1。系统开发。我们将构建一个以：i）新数值算法的VIM系统 重建3D体积图像； ii）一种新的轻弹性策略，以加快数据获取的速度； iii） 针对球形培养和药物治疗优化的微氟墨盒； iv）多色成像能力 分子检测。完整的VIM将解析构成球形的单个细胞以高分辨率 （横向，0.4 µm；轴向，1 µm）在大型成像体积中。 AIM 2。用肿瘤球体治疗监测。我们 将测试VIM的实际实用程序：启用VIM的球体成像比大量成像更早地揭示 球体反应敏感或对药物治疗的抗性。要生成肿瘤模型，我们将使用主要GBM 来自患者的细胞。 GBM球体将在微流体内生长并用药物（替莫唑胺）处理 墨盒。我们将使用VIM监控单个细胞在处理下如何改变其表型，并且 将这些变化与治疗结果相关联。影响。 VIM将成为癌症的变革工具 研究，通过丰富的数据集和高级分析赋予研究人员能力。即时 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，并在各种物理条件下前瞻性检测细胞命运。这些 结果将加强癌症研究中肿瘤球体的临床和科学效用。