FITC-DEXTRAN TRACERS IN MICRO-CIRCULATORY AND PERMEABILITY STUDIES USING COMBINED FLUORESCENCE STEREO MICROSCOPY, FLUORESCENCE LIGHT-MICROSCOPY AND ELECTRON-MICROSCOPY
FITC-DEXTRAN TRACERS IN MICRO-CIRCULATORY AND PERMEABILITY STUDIES USING COMBINED FLUORESCENCE STEREO MICROSCOPY, FLUORESCENCE LIGHT-MICROSCOPY AND ELECTRON-MICROSCOPY
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DOI：
10.1007/bf00507826
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发表时间：
1981-01-01
期刊：
影响因子：
--
通讯作者：
THORBALL, N
中科院分区：
文献类型：
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作者：
THORBALL, N
将荧光素-异硫氰酸酯偶联到葡聚糖（FITC-D）通过允许对组织样本进行荧光立体显微镜和高倍光学显微镜检查来扩展葡聚糖作为EM示踪颗粒的有用性。在实验期间、固定期间和随后的组织处理期间，可以在体内研究示踪剂的命运。还可以研究示踪剂的一些简单的物理化学特性，以及固定剂的影响（如果有的话）。FITC-D在6.5-8.5的pH范围内无变化，丙酮比乙醇沉淀更快，并在组织固定过程中以未知方式与戊二醛和四氧化锇反应。MW为70，000和150，000的FITC-D在用这些方法制备组织期间未显示扩散迹象，而当组织在固定剂载体中保存数天时，FITC-D 40，000在轻微程度上扩散。在组织固定和制备期间确保较低MW FITC-D的保存更加困难，并且所描述的方法是不充分的。右旋糖酐在大多数大鼠品系中引起过敏反应，但Wistar Furth大鼠耐受良好。将FITC引入右旋糖酐分子可能会改变生物反应，但Wistar Furth大鼠也耐受良好。组合的荧光立体显微镜，荧光显微镜的部分，染色切片和EM的光学显微镜，使得有可能遵循一个特定的微循环区域，在体内选择，在EM的最终研究。
Coupling fluorescein-isothiocyanate to dextrans (FITC-D) extends the usefulness of the dextrans as EM tracer particles by permitting preceding fluorescence stereomicroscopy and high-power light microscopy of the tissue specimens. The fate of the tracer may be studied in vivo during the experiment, during fixation and during the succeeding tissue processing. A study of some simple physicochemical characteristics of the tracer, and the influence, if any, of the fixing agent are also made possible. FITC-D was unchanged in the pH range from 6.5-8.5, more rapidly precipitated by acetone than by alcohol, and to react with glutaraldehyde and osmium tetroxide in an unknown way during tissue fixation. FITC-D with MW 70,000 and 150,000 showed no signs of diffusion during tissue preparation with these methods, whereas FITC-D 40,000 did so to a slight degree, when the tissue was kept for several days in the fixative vehicle. Securing the preservation of the lower MW FITC-D during tissue fixation and preparation is more difficult, and the described methods are not adequate. Dextrans provoke an anaphylactic reaction in most rat strains, but are well tolerated by Wistar Furth rats. The introduction of FITC into the dextran molecule might alter the biological reactions, but was also well tolerated by Wistar Furth rats. Combined fluorescence stereomicroscopy, fluorescence microscopy of sections, light microscopy of stained sections and EM made it possible to follow a particular microcirculatory area, selected in vivo, to the final study in the EM.