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Super-resolution fluorescence microscopy (SRFM) provides a better resolution than conventional fluorescence microscopy by reducing the point spread function (PSF). PSF is the light intensity distribution from a point that causes it to appear blurred. Due to PSF, each fluorescing point appears bigger than its actual size, and it is the PSF interference of nearby fluorophores that causes the blurred image. Various approaches to achieving higher resolution through SRFM have recently been...
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一个近红外Aptamer:活细胞超分辨率RNA成像的染料系统

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我们开发了SiRiuS, 一种用于活细胞RNA成像的近红外光吸收系统. 这种先进的探测器为超分辨率显微镜和动态RNA过程可视化提供了高性能.

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科学领域:

  • 分子生物学
  • 生物化学
  • 显微镜

背景情况:

  • 罗达胺 (SiR) 探针为活细胞和超分辨率成像提供了优势.
  • 光照明器 (FLAP) 是RNA可视化的新兴工具.
  • 在基于FLAP的RNA成像中,SiR探测器未得到充分利用.

研究的目的:

  • 开发一种基于SiR的高性能FLAP系统,用于哺乳动物活细胞中的RNA成像.
  • 增强阿普坦-染料相互作用,以提高性和光稳定性.
  • 实现动态RNA过程的时间分辨率和超分辨率成像.

主要方法:

  • 结合进化的体进化 (FACS) 与理性设计,突变和切断.
  • 有系统的化学衍生来改善SiR染料.
  • 在活细胞成像和STED超分辨率显微镜中进行验证.

主要成果:

  • 开发了具有高性和光稳定的SiRiuS:SiR-5染剂对.
  • 证明强烈的光增强有效的活细胞RNA可视化.
  • 启用了动态RNA过程的时间分辨率成像,例如压力颗粒的形成.
  • 在STED超分辨率显微镜中成功应用该系统用于亚衍射极限RNA成像.

结论:

  • SiRiuS:SiR-5系统是一个强大的NIR成像平台,用于活细胞中的RNA.
  • 它有助于先进的应用,包括超分辨率RNA显微镜.
  • 其与其他FLAP的正交性提高了其用于复杂RNA动态研究的实用性.