相关实验视频

Updated: Sep 13, 2025

Quantitative Microtubule Fractionation Technique to Separate Stable Microtubules, Labile Microtubules, and Free Tubulin in Mouse Tissues

Quantitative Microtubule Fractionation Technique to Separate Stable Microtubules, Labile Microtubules, and Free Tubulin in Mouse Tissues

Published on: November 17, 2023

基于单分子定位显微镜的微管超结构的定量表征

Zhao Xie¹, Fen Hu¹, Mingxin Chen¹

¹The Key Laboratory of Weak-Light Nonlinear Photonics of Education Ministry, School of Physics and TEDA Institute of Applied Physics, Nankai University, Tianjin, China.

Cytoskeleton (Hoboken, N.J.)

|July 30, 2025

概括

单分子局部化显微镜揭示了化学固定对微管完整性的影响. 甲-甲联合固定最好地保留了纳米尺度成像的微管结构.

科学领域:

细胞生物学细胞生物学
显微镜的使用方法
生物化学生物化学

背景情况:

传统的成像方法往往会掩盖细胞骨架构的纳米级细节.
单分子局部化显微镜 (SMLM) 为可视化超结构细节提供超分辨率能力.
微管的结构完整性对于细胞功能至关重要,并且可以受到固定协议的影响.

研究的目的:

开发一个定量框架,用SMLM来评估微管连续性和完整性.
评估各种化学固定协议对微管结构保存的影响.
为基于SMLM的微管成像和药理学查建立一个最佳的固定协议.

主要方法:

应用SMLM超分辨率成像来分析微管结构.
使用碎片化指数 (FI) 量化微管碎片化的计算算法开发.
用诺科达治疗的细胞中微管长度指数 (LI) 的定量分析.

主要成果:

与传统成像相反,SMLM揭示了显著的微管碎片化与甲 (PFA) 固定.
建立了固定效率的等级:4%的PFA > 甲醇 > 1%的甲 (GA) ≈3%的PFA + 0.1%的GA.
PFA-GA联合固定显示出优越的结构保存与最小的背景噪声,即使在短固定时间 (10分钟).
诺可达治疗显示了由LI量化的剂量依赖的微管分解.

结论:

基于SMLM的定量管道被开发用于评估微管道完整性.
准甲-甲联合固定被确定为SMLM中维护微管结构的最佳协议.
开发的框架作为一种可扩展的工具,用于基于超分辨率的微管向剂的药理学查.

关键词:

化学固定是化学固定的一种方式.计算算法计算算法免疫光成像成像技术微管是微管的组成部分.单分子定位显微镜 (SMLM) 技术超高分辨率成像成像技术

更多相关视频

Simultaneous Visualization of the Dynamics of Crosslinked and Single Microtubules In Vitro by TIRF Microscopy

Simultaneous Visualization of the Dynamics of Crosslinked and Single Microtubules In Vitro by TIRF Microscopy

Published on: February 18, 2022

Directly Measuring Forces Within Reconstituted Active Microtubule Bundles

Directly Measuring Forces Within Reconstituted Active Microtubule Bundles

Published on: May 10, 2022

相关概念视频

Microtubules

Microtubules

There are three types of cytoskeletal structures in eukaryotic cells—microfilaments, intermediate filaments, and microtubules. With a diameter of about 25 nm, microtubules are the thickest of these fibers. Microtubules carry out a variety of functions that include cell structure and support, transport of organelles, cell motility (movement), and the separation of chromosomes during cell division.

Studying the Cytoskeleton

Studying the Cytoskeleton

The cytoskeletal architecture can be studied using different microscopic and biochemical techniques. Electron microscopy was instrumental in discovering the cytoskeletal architecture around the 1960s, which allowed obtaining structural information at a high-resolution level. However, the sample preparation procedure often limits this ability in biological samples. Several protocols have been developed over the years to optimize sample preparation. In one of the protocols known as rotary...