Stereological and Flow Cytometry Characterization of Leukocyte Subpopulations in Models of Transient or Permanent Cerebral Ischemia
作者： Iván Ballesteros Martín , María Isabel Cuartero Desviat , Ana Moraga Yébenes , Juan de la Parra , Ignacio Lizasoaín Hernández
DOI: 10.3791/52031
关键词:
摘要: Microglia activation, as well extravasation of haematogenous macrophages and neutrophils, is believed to play a pivotal role in brain injury after stroke. These myeloid cell subpopulations can display different phenotypes functions need be distinguished characterized study their regulation contribution tissue damage. This protocol provides two methodologies for immune characterization: precise stereological approach flow cytometric analysis. The based on the optical fractionator method, which calculates total number cells an area interest (infarcted brain) estimated by systematic random sampling. second characterization simple way isolate leukocyte suspensions characterize them cytometry, allowing microglia, infiltrated monocytes neutrophils ischemic tissue. In addition, it also details cerebral ischemia model mice that exclusively affects cortex, generating highly reproducible infarcts with low rate mortality, procedure histological processing infarct volume Cavalieri method.
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