High and Low Molecular Weight Fluorescein Isothiocyanate (FITC)-Dextrans to Assess Blood-Brain Barrier Disruption: Technical Considerations.
High and Low Molecular Weight Fluorescein Isothiocyanate (FITC)-Dextrans to Assess Blood-Brain Barrier Disruption: Technical Considerations.
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DOI：
10.1007/s12975-010-0049-x
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发表时间：
2011-03
影响因子：
6.9
通讯作者：
Wintermark M
中科院分区：
文献类型：
--
作者：
Hoffmann A;Bredno J;Wendland M;Derugin N;Ohara P;Wintermark M
本文旨在报告组织学制备技术如何影响不同分子大小的异硫氰酸荧光素 (FITC)-葡聚糖的外渗模式，通常用作血脑屏障渗漏的标记物。通过使用适当的制备方法，可以最大限度地减少假阴性结果。 Wistar 大鼠接受了 2 小时的大脑中动脉闭塞和磁共振成像。最后一次成像扫描后，注射伊文思蓝和分子量为 4、40 和 70 kDa 的 FITC-葡聚糖。使用不同的组织学制备方法。通过荧光显微镜分析血脑屏障渗漏部位。使用所有制备方法都可以检测到梗塞区域伊文思蓝和高分子 FITC-葡聚糖（40 和 70 kDa）的外渗。如果直接暴露于盐水，这些 FITC-葡聚糖的信号强度会降低。 4-kDa 低分子量 FITC-葡聚糖的外渗只能使用新鲜冷冻的组织切片进行检测。涉及多聚甲醛和蔗糖的制剂导致 4-kDa FITC-葡聚糖溶解在这些反应物中并被洗掉，从而产生无外渗的假阴性结果。 FITC-葡聚糖是表征动物模型中血脑屏障通透性改变的重要工具。通过立即冷冻组织进行直接固定，可以避免低分子量 FITC-葡聚糖的扩散和洗掉。需要了解这一陷阱，以避免产生低分子量 FITC-葡聚糖没有外渗的错误印象。
This note is to report how histological preparation techniques influence the extravasation pattern of the different molecular sizes of fluorescein isothiocyanate (FITC)–dextrans, typically used as markers for blood-brain barrier leakage. By using appropriate preparation methods, false negative results can be minimized. Wistar rats underwent a 2-h middle cerebral artery occlusion and magnetic resonance imaging. After the last imaging scan, Evans blue and FITC–dextrans of 4, 40, and 70 kDa molecular weight were injected. Different histological preparation methods were used. Sites of blood-brain barrier leakage were analyzed by fluorescence microscopy. Extravasation of Evans blue and high molecular FITC–dextrans (40 and 70 kDa) in the infarcted region could be detected with all preparation methods used. If exposed directly to saline, the signal intensity of these FITC–dextrans decreased. Extravasation of the 4-kDa low molecular weight FITC–dextran could only be detected using freshly frozen tissue sections. Preparations involving paraformaldehyde and sucrose resulted in the 4-kDa FITC–dextran dissolving in these reactants and being washed out, giving the false negative result of no extravasation. FITC–dextrans represent a valuable tool to characterize altered blood-brain barrier permeability in animal models. Diffusion and washout of low molecular weight FITC–dextran can be avoided by direct immobilization through immediate freezing of the tissue. This pitfall needs to be known to avoid the false impression that there was no extravasation of low molecular weight FITC–dextrans.
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影响因子：
3.6
作者：
EMMRICH, P;BAUMANN, W;STECHELE, U
通讯作者：
STECHELE, U
DOI：
10.1007/bf00507826
发表时间：
1981-01-01
期刊：
HISTOCHEMISTRY
影响因子：
--
作者：
THORBALL, N
通讯作者：
THORBALL, N
影响因子：
12.7
作者：
WOLMAN, M;KLATZO, I;SPATZ, M
通讯作者：
SPATZ, M
影响因子：
6.3
作者：
Knight, Robert A.;Karki, Kishor;Nagaraja, Tavarekere N.
通讯作者：
Nagaraja, Tavarekere N.
影响因子：
2.4
作者：
Nagaraja, Tavarekere N.;Keenan, Kelly A.;Knight, Robert A.
通讯作者：
Knight, Robert A.