癌症研究
活细胞成像在
癌症研究中的使用
癌症研究基本概述
癌症的特点是一种复杂的疾病，由一系列连续的基因突变引起，导致细胞功能和增殖缺陷。 新科学技术的发展以及现有方法的优化推动了癌症研究的进步。
活细胞显微镜已成为提高我们对癌症发病机制理解的不可或缺的工具。 延时成像的应用范围从直接可视化免疫细胞和癌细胞之间的瞬时相互作用，到检查更复杂的生物过程，如转移、程序性细胞死亡和肿瘤血管生成。
用于评估抗癌剂的细胞毒性和基因毒性的多板克隆测定
细胞存活和集落形成的无标记、半自动评估
克隆形成或集落形成测定是癌症生物学领域中广泛使用的方法。 它评估了不同治疗对癌细胞存活和形成集落能力的影响。 标准的克隆形成试验方案包括在漫长的潜伏期（1-3 周）结束时检查新形成的菌落。 为了使菌落可视化，需要固定细胞并用 DNA 嵌入染料结晶紫染色。 CytoSMART Omni 通过创建高质量的延时视频，一次持续数天甚至数周，而无需修复或染色您的细胞，从而可以连续监测克隆生长。 监测集落形成的整个过程可以提供很多有价值的信息，包括不同药物治疗的时间依赖性效应。 此外，集成的图像分析软件会自动检测新形成的菌落并评估它们的大小和圆形度。
荧光活细胞显微镜在癌症免疫治疗中的应用
细胞间相互作用的标记成像
癌症免疫疗法利用宿主免疫系统的固有能力来对抗癌症。 荧光显微镜在促进了解免疫细胞和肿瘤细胞之间发生的相互作用方面发挥着关键作用，因为这种细胞相互作用代表了任何免疫治疗方法的核心。 CytoSMART Lux3 FL 荧光成像系统可以通过实时收集动力学数据来捕捉不同类型细胞之间的空间相互作用。 由于成像可以直接从培养箱内部进行，细胞相互作用不会受到不想要的外在因素的影响。
用于研究癌症转移的体外细胞迁移试验
基于算法的细胞迁移分析
癌症转移涉及恶性细胞从其原始部位扩散，是导致癌症相关死亡人数最多的原因。 已经开发了许多体外测定以在更受控的环境下复制转移的不同阶段。 划痕分析（也称为伤口愈合分析）尤其可以帮助分析在各种趋化因子和生长因子影响下的癌细胞迁移。 所有 CytoSMART 活细胞成像平台（CytoSMART Omni、Lux2 或 Lux3FL）都配备了划痕试验的软件，可以分析划痕闭合和细胞迁移的速率和效率。
用于癌症建模的 3D 细胞培养
肿瘤类器官代表了一种更复杂的癌症研究模型，因为它们更准确地模拟了癌症的遗传、细胞和病理生理特征。 CytoSMART 技术已经开发出有效的解决方案来处理肿瘤衍生的类器官。 CytoSMART Omni 明场成像系统可以通过直接从培养箱内记录全血管延时视频来密切监测肿瘤类器官的形成过程。 康宁细胞计数器采用创新的类器官计数软件，可以立即确定样品中类器官的精确数量和大小。
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Appnotes
Cell Cytoxicity Assay to Analyze Drug Response
Cell Viability Analysis
Cell Cytoxicity Assay to Analyze Drug Response
The effect of drugs and drug candidates on the viability of cells in culture can be determined using cell counting, live/dead assays and metabolic assays. However, these assays are often end-point measurements. Alternatively, cells can be monitored using bright-field microscopy, by creating time-lapse videos for a culture period of multiple days. To study the lasting effect of the drug candidate.
In this study the cytotoxic effect of Paclitaxel, a chemotherapy drug, was investigated for a range of concentrations. The effect on cell viability between drug concentrations was compared by analyzing confluency measurements obtained using automated live-cell imaging. The entire experiment was performed inside a CO2-incubator, ensuring optimal culturing conditions and cells were imaged every hour for a period of 3 days.
Cell Migration
Cell migration is essential for physiological development and homeostasis, among other things. It is part of processes such as angiogenesis and wound healing. Conversely, cell migration in pathologies, including cancer can lead to worsening and progression of the diseased state.
To gain insight into collective cell migration a variety of assays have been developed. One of which is the wound healing assay, also known as the scratch assay. In this procedure a ‘wound’ is made in a confluent monolayer of cells, after which the gap closure is quantitatively monitored.
For this study a wound healing assay was performed to assess the effect of the drug paclitaxel on the migration of C6 rat glail tumor cells. Automated live-cell imaging was performed inside a CO2-incubator to ensure cells are kept at the desired conditions during the entire experiment. The experiment was performed in a 24 well plate, which was fully imaged every hour for 23 hours. The surface area and gap closure speed were compared for a concentration range of paclitaxel.
Cell Viability Analysis
Cell viability, growth and cytotoxicity studies can be performed using metabolic activity assays. The overall metabolic activity of the cell is indicated by the enzymatic cleavage of colorimetric or fluorescent substrates.
While these assays are relatively straightforward and cheap, they are dependent on culture conditions and intrinsic metabolic activity of the cell type that is being investigated. Furthermore, depletion of the metabolic substrate can lead to a plateau in the fluorescent signal, making assay output unreliable. To overcome these limitations cell viability could be determined optically using confluency measurements.
In the study described here the performance of confluency measurements to assess cell viability were compared to a metabolic activity assay: cell titer blue. Confluency was visualized using automated bright-field microscopy and subsequently analyzed using image analysis algorithms. Images were collected inside a CO2-incubator, keeping the culture at optimal conditions. For the cell titer blue assay resazurin was added to the medium and incubated for 3 hours. The fluorescent signal was normalized to the control to obtain the relative metabolic activity as a measure of cell viability. The comparison between the methods was performed for two pancreatic cancer cell lines, PACO7 and POCA43.