• 主题
高级搜索  >
历史搜索 清空历史
首页> 外文会议>Conference on Multiphoton Microscopy in the Biomedical Sciences IV; 20040125-20040127; San Jose,CA; US >Multiphoton FLIM and Spectral Imaging of Cells and Tissues
【24h】

Multiphoton FLIM and Spectral Imaging of Cells and Tissues

机译：细胞和组织的多光子FLIM和光谱成像

获取原文
获取原文并翻译 | 示例
页面导航
  • 摘要
  • 著录项
  • 相似文献
  • 相关主题

摘要

Five-dimensional (5D) multiphoton measurements with submicron spatial resolution, 270 ps temporal resolution and 5 nm spectral resolution have been performed on living cells and tissues at 750 nm - 850 nm laser excitation. A compact (65x62x48 cm~3) multiport laser scanning microscope TauMap (JenLab GmbH) equipped with fast PMT and CCD camera, SPC 830 time-correlated single photon counting board and Sagnac interferometer was used. Laser excitation radiation was provided by a tuneable MaiTai Thsapphire femtosecond laser as well as by a 405 nm 50 MHz picosecond laser diode. The spectral and temporal fluorescence behaviour of intratissue chloroplasts of water plant leafs, of a variety of exogenous fluorophores as well as of fluorescent proteins in transfected brain cells have been studied. When calculating fluorescence lifetime images (FLIM) we found differences in intracellular two-photon fluorescence lifetimes vs. one-photon fluorescence lifetimes. Multiphoton FLIM-FRET and multiphoton spectral FRET studies have been performed in living HBMEC brain cells using CFP and YFP fusion proteins. It was shown that FLIM-FRET data depend on laser power due to photodestructive multiphoton effects. This has to be considered in long-term fluorescence resonance energy transfer studies of dynamic protein-protein interactions.
机译：已在750 nm-850 nm激光激发下对活细胞和组织进行了亚微米空间分辨率，270 ps时间分辨率和5 nm光谱分辨率的五维（5D）多光子测量。使用紧凑型（65x62x48 cm〜3）多端口激光扫描显微镜TauMap（JenLab GmbH），该显微镜配备了快速PMT和CCD相机，SPC 830时间相关的单光子计数板和Sagnac干涉仪。激光激发辐射由可调节的MaiTai Thsapphire飞秒激光器以及405 nm 50 MHz皮秒激光二极管提供。研究了水生植物叶片内叶绿体，各种外源荧光团以及转染的脑细胞中荧光蛋白的光谱和时间荧光行为。在计算荧光寿命图像（FLIM）时，我们发现细胞内两光子荧光寿命与单光子荧光寿命之间存在差异。已经使用CFP和YFP融合蛋白在活的HBMEC脑细胞中进行了多光子FLIM-FRET和多光子光谱FRET研究。结果表明，由于光破坏性多光子效应，FLIM-FRET数据取决于激光功率。在动态蛋白质-蛋白质相互作用的长期荧光共振能量转移研究中必须考虑到这一点。

著录项

相似文献

  • 外文文献
  • 中文文献
  • 专利
获取原文

客服邮箱：kefu@zhangqiaokeyan.com

京公网安备：11010802029741号 ICP备案号：京ICP备15016152号-6 六维联合信息科技 (北京) 有限公司©版权所有

  • 客服微信

  • 服务号