整合素依赖细胞矩阵粘附的成像和定量分析
Anne-Marieke D van Stalborch1, Andrew G Clark2, Arnoud Sonnenberg3
1Amsterdam University Medical Center, 1066 CX Amsterdam, the Netherlands.
STAR protocols
|August 24, 2023
概括
该协议详细描述了成像和分析因特林依赖细胞矩阵粘附,这对于诸如伤口愈合等各种生物过程至关重要. 它为研究细胞粘附动态的研究人员提供了全面的指南.
科学领域:
- 细胞生物学 细胞生物学
- 生物物理学的生物物理.
背景情况:
- 综合素介导的细胞-细胞外基质 (ECM) 粘附对于包括胚胎发育,免疫反应,组织组织和伤口愈合在内的基本生物过程至关重要.
- 了解这些粘附的动态和调节对于破译健康和疾病中的细胞行为至关重要.
研究的目的:
- 提出一个详细的方案,用于成像和定量分析的整合素依赖的细胞矩阵粘附.
- 为研究细胞-ECM相互作用提供从细胞培养到先进显微镜和数据分析的全面工作流.
主要方法:
- 该协议涵盖细胞培养,病毒制备和lentiviral转导用于细胞标签.
- 它详细介绍了成像技术,包括宽场,共聚焦和全内部反射光显微镜 (TIR-FM).
- 定量分析通过提供脚本进行,并使用活细胞成像和光漂白后光恢复 (FRAP) 评估粘附动力学.
主要成果:
- 该协议可以对细胞矩阵粘附进行强大的成像.
- 定量分析工具和FRAP方法允许对粘附尺寸,数量和动态进行详细评估.
- 提出的方法有助于研究积分依赖的粘附周转率和稳定性.
结论:
- 该协议为研究人员提供了一种标准化和全面的方法,用于研究整合素依赖的细胞矩阵粘附.
- 描述的技术对于我们进一步了解细胞粘附在生理和病理过程中的作用至关重要.
- 该协议支持对粘附动态的定量分析,这对于发育生物学和再生医学等领域至关重要.
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