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相关概念视频

Protein Dynamics in Living Cells01:19

Protein Dynamics in Living Cells

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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...
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Super-resolution Fluorescence Microscopy01:37

Super-resolution Fluorescence Microscopy

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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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A Guide to Structured Illumination TIRF Microscopy at High Speed with Multiple Colors
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通过结构化照明显微镜进行像素智能扩散成像,在光漂白后进行光恢复 (SIM-FRAP).

Dustin M Harmon1, Maura M Wimsatt1, Michelle L Cousineau2

  • 1Department of Chemistry, Purdue University, West Lafayette, Indiana 47907, United States.

Analytical chemistry
|February 16, 2026
PubMed
概括

结构化照明显微镜 - - 光漂白后光恢复 (SIM-FRAP) 能够在无形固体分散中进行像素式扩散映射. 这种技术量化了溶解前沿的分子运动,揭示了配方设计的洞察力.

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Internalization and Observation of Fluorescent Biomolecules in Living Microorganisms via Electroporation

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科学领域:

  • 制药科学 制药科学
  • 材料科学 材料科学 材料科学
  • 显微镜的使用方法

背景情况:

  • 在无形固体分散 (ASD) 中量化分子扩散性对于设计有效的药物配方至关重要.
  • 由于相分离,ASD中的高药物负载可以降低溶解率,形成富含药物的屏障层.
  • 分子移动性是了解紧体内的不同阶段溶解趋势的关键.

研究的目的:

  • 开发和应用一种新的SIM-FRAP技术,用于药品紧包中的像素智能扩散映射.
  • 研究ASD溶解前线的分子移动性,包括玻璃,水合,凝和相隔域.
  • 为了证明SIM-FRAP对分子扩散的无细分量化分析的能力.

主要方法:

  • 整合结构化照明显微镜 (SIM) 重建算法与光漂白 (FT-FRAP) 后的空间里叶变换光恢复.
  • 使用SIM进行光漂白,然后进行空间里叶变换和逆里叶变换,以生成依赖调制的振幅图.
  • 执行像素智能适合指数衰变函数,以确定整个视野的扩散系数.

主要成果:

  • 通过SIM-FRAP,SIM-FRAP成功地通过模拟制药紧件的溶解前端生成了分子移动性的像素智能扩散图.
  • 该技术揭示了紧和凝阶段平稳变化的扩散性,以及相隔域中不连续的变化.
  • 每个实验都能回收大量的扩散系数 (高达100万),提供详细的空间信息.

结论:

  • SIM-FRAP提供了一种强大的,没有细分的方法来量化ASD中的分子扩散性.
  • 该技术直接可视化分子移动性的空间变化,与假设的相位分离现象一致.
  • SIM-FRAP的实验简单性促进了其在配方开发和材料表征方面的更广泛应用.