Synchrotron UV Fluorescence Microscopy Uncovers New Probes in Cells and Tissues
Synchrotron UV Fluorescence Microscopy Uncovers New Probes in Cells and Tissues
复制标题
DOI：
10.1017/s1431927610093852
复制
发表时间：
2010-10-01
影响因子：
2.8
通讯作者：
Refregiers, Matthieu
中科院分区：
文献类型：
--
作者：
Jamme, Frederic;Villette, Sandrine;Refregiers, Matthieu
深紫外（DUV，低于350 nm）荧光的使用开辟了生物学的新可能性，因为它不需要外部特异性探针或标记，而是允许使用在该波长范围内激发时存在于许多生物分子中的固有荧光。事实上，观察无标记的生物分子或活性药物确保了标记不会改变生物定位或其任何性质。在过去，由于有限的光源和显微镜光学器件，实现DUV荧光成像并不容易。两个世界共存：具有DUV激发的全光谱信息的荧光分光光度测量，其缺乏高分辨率定位，以及具有非常好的空间分辨率但光谱分辨率差的微观世界，其波长范围限于350 nm。为了将两个世界的优势联合收割机结合起来，我们开发了一种用于细胞生物学的DUV荧光显微镜，它与同步加速器光束线耦合，提供180至600 nm的精细可调激发和在图像的每个点上获得的全光谱，以研究直接从活细胞或组织活检内的纳米体积发出的DUV激发荧光。
Use of deep ultraviolet (DUV, below 350 nm) fluorescence opens up new possibilities in biology because it does not need external specific probes or labeling but instead allows use of the intrinsic fluorescence that exists for many biomolecules when excited in this wavelength range. Indeed, observation of label free biomolecules or active drugs ensures that the label will not modify the biolocalization or any of its properties. In the past, it has not been easy to accomplish DUV fluorescence imaging due to limited sources and to microscope optics. Two worlds were coexisting: the spectrofluorometric measurements with full spectrum information with DUV excitation, which lacked high-resolution localization, and the microscopic world with very good spatial resolution but poor spectral resolution for which the wavelength range was limited to 350 nm. To combine the advantages of both worlds, we have developed a DUV fluorescence microscope for cell biology coupled to a synchrotron beamline, providing fine tunable excitation from 180 to 600 nm and full spectrum acquired on each point of the image, to study DUV excited fluorescence emitted from nanovolumes directly inside live cells or tissue biopsies.