title: Confocal imaging of microglial cell dynamics in hippocampal slice cultures.
abstract: Methods are described for imaging the cellular dynamics of microglia in live mammalian brain slice cultures. Brain slices prepared from developing rat hippocampus are cultured for up to 2 weeks by the roller tube or static filter culture technique, stained with one or more fluorescent dyes, and imaged by scanning laser confocal microscopy. One of several cell type-specific or nonspecific fluorescent dyes can be used independently or in combination to label cells in live brain tissues. The fluorescently conjugated plant isolectin GSA-IB4 is useful for identifying microglia and for following their structure, movement, and proliferation. Live and dead neurons and glia can be distinguished using membrane-permeant and -impermeant fluorescent nucleic acid dyes. Nonspecific fluorescent lipids such as DiIC18 can be used as a vital stain to label populations of endocytic and phagocytic cells. Using multichannel confocal imaging, tissue slices that are single-, double-, or triple-labeled can be imaged in the living state in two or three spatial dimensions as well as in time. This provides a means for investigating the cell-cell interaction and dynamic behavior of microglia and other cell types in live brain tissues cultured under various physiological conditions.
authors.affiliation: Department of Biological Sciences, University of Iowa, Iowa City, Iowa 52242-1324, USA. michael-e-dailey@uiowa.edu
authors.forename: M E
authors.initials: ME
authors.lastname: Dailey
authors.forename: M
authors.initials: M
authors.lastname: Waite
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D004210:Dissection / Q000379:methods; D004705:Endocytosis; D016650:Fluorescein-5-isothiocyanate; D005456:Fluorescent Dyes; D006624:Hippocampus / Q000166:cytology* / Q000502:physiology*; D037102:Lectins; D017628:Microglia / Q000166:cytology* / Q000502:physiology*; D018613:Microscopy, Confocal / Q000379:methods; D009457:Neuroglia / Q000166:cytology; D009474:Neurons / Q000166:cytology; D009924:Organ Culture Techniques / Q000295:instrumentation / Q000379:methods; D051381:Rats; D012377:Rodentia
chemical_list: D005456:Fluorescent Dyes; D037102:Lectins; D016650:Fluorescein-5-isothiocyanate