Stracke Frank, Kreiner-Møller Asger, Zimmermann Heiko
Fraunhofer Institute for Biomedical Technology, Ensheimer Str. 48, 66386, St. Ingbert, Germany.
Methods Mol Biol. 2015;1257:229-41. doi: 10.1007/978-1-4939-2193-5_8.
Laser scanning microscopy is emerging as a powerful imaging tool in cryobiology. The basic microscopy system can be combined with various imaging modalities including Raman spectroscopy, fluorescence correlation spectroscopy, fluorescence lifetime imaging, or multiphoton imaging. Multiphoton imaging can be used to study intracellular ice formation at the subcellular level. A Raman imaging modality can be used for chemical mapping of frozen samples. A Raman spectrum gives information about characteristic molecular vibrations of specific groups in molecules. Raman images can be used to determine the localization of intra- and extracellular constituents and the various forms of water in freeze-concentrated solutions. Spectra can be collected during freezing and thawing of a sample using a temperature-controlled sample holder. In this chapter, various advanced cryoimaging methods are described. Special emphasis is given on the different imaging modalities that can be used to study the various aspects of cryopreservation.
激光扫描显微镜正成为低温生物学中一种强大的成像工具。基本的显微镜系统可以与各种成像方式相结合，包括拉曼光谱、荧光相关光谱、荧光寿命成像或多光子成像。多光子成像可用于在亚细胞水平研究细胞内冰晶形成。拉曼成像方式可用于对冷冻样品进行化学绘图。拉曼光谱给出了分子中特定基团的特征分子振动信息。拉曼图像可用于确定冷冻浓缩溶液中细胞内和细胞外成分以及各种水形态的定位。使用温度可控的样品架可以在样品冷冻和解冻过程中收集光谱。在本章中，描述了各种先进的低温成像方法。特别强调了可用于研究冷冻保存各个方面的不同成像方式。
