Document Detail


High and Low Molecular Weight Fluorescein Isothiocyanate (FITC)-Dextrans to Assess Blood-Brain Barrier Disruption: Technical Considerations.
MedLine Citation:
PMID:  21423333     Owner:  NLM     Status:  PubMed-not-MEDLINE    
Abstract/OtherAbstract:
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.
Authors:
Angelika Hoffmann; Jörg Bredno; Michael Wendland; Nikita Derugin; Peter Ohara; Max Wintermark
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Publication Detail:
Type:  Journal Article     Date:  2010-11-11
Journal Detail:
Title:  Translational stroke research     Volume:  2     ISSN:  1868-4483     ISO Abbreviation:  Transl Stroke Res     Publication Date:  2011 Mar 
Date Detail:
Created Date:  2013-12-10     Completed Date:  2013-12-11     Revised Date:  2013-12-12    
Medline Journal Info:
Nlm Unique ID:  101517297     Medline TA:  Transl Stroke Res     Country:  United States    
Other Details:
Languages:  eng     Pagination:  106-11     Citation Subset:  -    
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