自动化FRAP显微镜用于高通量分析染色体组织和转录中的蛋白质动态
Selçuk Yavuz1, Bart Geverts2, Johan A Slotman2
1Department of Pathology, Erasmus University Medical Center, Rotterdam, The Netherlands.
FEBS open bio
|December 6, 2025
概括
RoboMic是一个自动化显微镜平台,增强了用于蛋白质动力学研究的高通量细胞成像. 它使用人工智能和并行成像来显著增加光漂白后光恢复 (FRAP) 测量.
科学领域:
- 细胞和分子生物学 细胞和分子生物学
- 生物物理学的生物物理.
- 显微镜和成像技术技术.
背景情况:
- 光漂白后光恢复 (FRAP) 对于研究活细胞中的蛋白质动态至关重要.
- 现有的FRAP方法需要人工干预,限制了吞吐量和可扩展性.
- 高通量定量成像对于大规模的生物研究至关重要.
研究的目的:
- 开发RoboMic,一个完全自动化的共聚焦显微镜平台,用于高通量FRAP测试.
- 引入并行FRAP (pFRAP) 进行同时的多细胞分析,显著提高效率.
- 为了证明RoboMic在为蛋白质移动性研究生成可靠数据的能力.
主要方法:
- 开发自动化显微镜控制软件,使用人工智能驱动的细胞核选择和细分.
- 实施顺序FRAP (sFRAP) 和并行FRAP (pFRAP) 成像协议.
- 感兴趣的亚核区域 (ROI) 的自动定义,光漂白和时间间隔成像.
主要成果:
- 使用pFRAP,RoboMic在保持分辨率的同时实现了5到10倍的吞吐量增加.
- 基于人工智能的自动化分析使得FRAP测量效率高且可重复.
- 证明了对雄激素受体和凝聚素复合体的核动力学的强大数据生成.
结论:
- RoboMic显著推进了研究蛋白质动态的高通量定量成像.
- 该平台增加了大规模蛋白质流动性研究的统计能力和可扩展性.
- RoboMic可以适应FRAP之外的各种定量功能成像测试.
相关概念视频
01:19Protein Dynamics in Living Cells
2.6K
Different fluorescence-based techniques are used to study the protein dynamics in living cells. These techniques include FRAP, FRET, and PET.
Fluorescent recovery after photobleaching (FRAP) is a fluorescent-protein-based detection technique used to quantify protein movement rates within the cell. This method exposes a small portion of the cell to an intense laser beam. The laser beam causes permanent photobleaching of the fluorophore-tagged proteins in the exposed region. As the bleached...
Fluorescent recovery after photobleaching (FRAP) is a fluorescent-protein-based detection technique used to quantify protein movement rates within the cell. This method exposes a small portion of the cell to an intense laser beam. The laser beam causes permanent photobleaching of the fluorophore-tagged proteins in the exposed region. As the bleached...
2.6K
02:36Chromatin Immunoprecipitation- ChIP
12.1K
Chromatin immunoprecipitation, or ChIP, is an antibody-based technique used to identify sites on DNA that bind to transcription factors of interest or histone proteins. It also helps determine the type of histone modifications such as acetylation, phosphorylation, or methylation.
Types of ChIP
ChIP can be divided into two types - X-ChIP and N-ChIP. X-ChIP involves in vivo cross-linking of histones and regulatory proteins to DNA, fragmenting the DNA by sonication, and isolating the protein-DNA...
Types of ChIP
ChIP can be divided into two types - X-ChIP and N-ChIP. X-ChIP involves in vivo cross-linking of histones and regulatory proteins to DNA, fragmenting the DNA by sonication, and isolating the protein-DNA...
12.1K