| Literature DB >> 25502884 |
Thomas Wälchli1, José María Mateos2, Oliver Weinman3, Daniela Babic4, Luca Regli5, Simon P Hoerstrup6, Holger Gerhardt7, Martin E Schwab3, Johannes Vogel8.
Abstract
During development and in various diseases of the CNS, new blood vessel formation starts with endothelial tip cell selection and vascular sprout migration, followed by the establishment of functional, perfused blood vessels. Here we describe a method that allows the assessment of these distinct angiogenic steps together with antibody-based protein detection in the postnatal mouse brain. Intravascular and perivascular markers such as Evans blue (EB), isolectin B4 (IB4) or laminin (LN) are used alongside simultaneous immunofluorescence on the same sections. By using confocal laser-scanning microscopy and stereological methods for analysis, detailed quantification of the 3D postnatal brain vasculature for perfused and nonperfused vessels (e.g., vascular volume fraction, vessel length and number, number of branch points and perfusion status of the newly formed vessels) and characterization of sprouting activity (e.g., endothelial tip cell density, filopodia number) can be obtained. The entire protocol, from mouse perfusion to vessel analysis, takes ∼10 d.Entities:
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Year: 2014 PMID： 25502884 DOI： 10.1038/nprot.2015.002
Source DB: PubMed Journal: Nat Protoc ISSN： 1750-2799 Impact factor: 13.491