Quantitative phase imaging microscopy for label-free, high-content imaging

光学显微镜是生物医学研究中的一种常规工具，它允许我们观察细胞的内部工作方式，以及细胞如何共同形成组织。能够看到细胞的运动有助于我们了解健康细胞和组织中的重要过程以及疾病中的问题所在。细胞本质上是透明的，很难在其自然状态下成像。通常，我们添加荧光标记来实现细胞亚结构或特定蛋白质的可视化。虽然这样的荧光探针可以产生出色的高对比度图像，但探针的引入和成像都可能对细胞活性有害，并干扰观察过程。例如，一个庞大的荧光分子与感兴趣的蛋白质结合，可以影响该分子的行为，而荧光成像会产生对细胞有害的自由基。在一些细胞中，如患者来源的细胞或其他敏感细胞类型，可能根本不可能添加荧光标记，使得荧光成像变得不可能。透过透明细胞的光经历相移，通过用参考光束干涉这种光，我们可以产生全息图。使用精心设计的算法，可以从这些全息图中提取细胞密度形式的空间信息。从这些全息图产生的图像类似于从荧光标记的细胞获得的图像，提供不需要荧光标记的高对比度图像。这项技术被称为定量相位成像，允许我们以最小的扰动对细胞进行自然状态的成像。这个应用程序请求资金购买定量相位成像显微镜，用于微创细胞成像。所要求的显微镜将配备全自动采集和分析，使我们能够并行成像多达96个样本，并以客观的方式自动提取有关细胞增殖、运动和形状的信息。因此，这种显微镜将支持一系列发现和转译研究项目的基于细胞的筛选，以了解和开发罕见遗传病、细菌感染和癌症的治疗方法。

Optical microscopy is a routine tool in biomedical research, allowing us to view the inner workings of cells and how cells work together to form tissues. Being able to see the movement of cells helps us to understand important processes in healthy cells and tissues and what goes wrong in disease.Cells are inherently transparent and difficult to image in their native state. Typically, we add fluorescent markers to enable visualisation of cellular substructures or specific proteins. While such fluorescent probes result in excellent high contrast images, both the introduction and imaging of the probe can be detrimental to cell viability and disturb the observed process. The binding of a bulky fluorescent molecule to a protein of interest, for example, can affect the behaviour of that molecule, while fluorescence imaging produces free radicals which are harmful to the cells. In some cells, such as patient-derived cells or other sensitive cell types, it may not be possible to add fluorescent markers at all, making fluorescence imaging impossible.Light transmitted through transparent cells undergoes a phase shift, by interfering this light with a reference beam, we can generate holograms. Spatial information in the form of cellular density can be extracted from these holograms using carefully designed algorithms. The images generated from these holograms resemble those achieved from fluorescently labelled cells, supplying high contrast images without requiring fluorescent markers. This technique, known as quantitative phase imaging, allows us to image cells in their native state with minimal perturbation.This application requests funds to purchase a quantitative phase imaging microscope for minimally invasive cellular imaging. The requested microscope will come with fully automated acquisition and analysis, enabling us to image up to 96 samples in parallel and extract information on the proliferation, movement and shape of cells automatically in an objective manner. This microscope will therefore support cell-based screening for a range of discovery and translational research projects to understand and develop treatments for rare genetic diseases, bacterial infections and cancer.