活细胞表面的小分子标记用于三维超高分辨率显微镜
Marissa K Lee1, Prabin Rai, Jarrod Williams
1Department of Chemistry, Stanford University , Stanford, California 94305, United States.
Journal of the American Chemical Society
|September 16, 2014
概括
我们开发了一种新的罗达胺螺旋乳染料,用于对细菌细胞表面进行无毒,超分辨率 (SR) 的成像. 这种可光控制的发射器可以通过可见光精确地可视化细胞结构,从而为细胞生长提供了新的见解.
科学领域:
- 显微镜和成像技术
- 化学生物学 化学生物学
- 细菌细胞生物学 细菌细胞生物学
背景情况:
- 由于尺寸限制,细菌细胞表面成像需要超分辨率方法.
- 传统的罗达胺螺旋使用有害的紫外线进行光交换.
研究的目的:
- 开发一种可光控制的罗达胺螺旋,用于无毒的3D超分辨率 (SR) 成像.
- 为了能够对活细菌细胞表面进行特定的标记.
主要方法:
- 代合成和光谱鉴定罗达胺螺旋乳酸盐.
- 功能化与N-hydroxysuccinimide用于共价胺标签.
- 3D超分辨率 (SR) 显微镜对标记的Caulobacter crescentus进行检测.
主要成果:
- 优化了可见光 (>400 nm) 的罗达螺旋胺光开关.
- 共价标签实现了细菌细胞表面的统一和特异性采样.
- 高分辨率 (10-20纳米) 3D SR重建揭示了细胞茎长度分布和生长部位.
结论:
- 可光控制的罗达胺螺旋乳为活细胞SR成像提供了一种无毒的,特定的标记策略.
- 这种方法允许细菌表面动态和结构的详细研究.
- 优化的染料是纳米生物成像的多功能工具.
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