通过在单分子定位显微镜中限制受限生物分子静态运动来提高定位精度
Jielei Ni1, Bo Cao1,2, Gang Niu3,4,5,6
1Nanophotonics Research Center, Shenzhen Key Laboratory of Micro-Scale Optical Information Technology and Institute of Microscale Optoelectronics, College of Physics and Optoelectronic Engineering, College of Electronics and Information Engineering, Shenzhen University, Shenzhen 518060, China.
Nanophotonics (Berlin, Germany)
|December 5, 2024
概括
固定生物分子的有限静态运动阻碍了单分子定位显微镜 (SMLM) 中的定位精度. 例如,通过后交联来限制这种运动,可以显著提高SMLM成像的准确性.
科学领域:
- 生物物理学的生物物理.
- 显微镜的使用方法
- 分子生物学分子生物学
背景情况:
- 单分子定位显微镜 (SMLM) 对于解决超出衍射极限的纳米尺寸生物分子至关重要.
- 预计SMLM中的固定生物分子会表现出有限的随机运动,但其对定位精度的影响尚不清楚.
研究的目的:
- 调查局限随机运动在SMLM中的定位精度的检测能力和影响.
- 探索通过管理生物分子运动来提高定位精度的方法.
主要方法:
- 在不同的间隔下分析固定的Alexa Fluor-647-结合的寡核酸的局部位移.
- 计算生物分子运动与定位精度之间的关系.
- 评估生物分子大小和后交叉连接对精度的影响,使用染色为氨酸的细胞样本.
主要成果:
- 生物分子的随机运动被检测到并被发现是受限的,随着时间间隔的增加而增加的位移,然后才能和.
- 发现定位精度与局限性随机运动的程度成反比例.
- 放大生物分子降低了精度,而后交叉连接则提高了精度,如图布林成像所示.
结论:
- 固定生物分子的有限静态运动会对SMLM定位精度产生负面影响.
- 限制这种运动的策略,例如后交叉链接,可以提高SMLM中的成像准确性.
相关概念视频
01:37Super-resolution Fluorescence Microscopy
6.9K
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...
6.9K
01:19Protein Dynamics in Living Cells
2.1K
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...
2.1K