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Titel: The in situ morphology of microglia is highly sensitive to the mode of tissue fixation
VerfasserIn: Cătălin, Bogdan
Stopper, Laura
Bălşeanu, Tudor-Adrian
Scheller, Anja
Sprache: Englisch
Titel: Journal of Chemical Neuroanatomy
Bandnummer: 86
Seiten: 59-66
Verlag/Plattform: Elsevier
Erscheinungsjahr: 2017
DDC-Sachgruppe: 610 Medizin, Gesundheit
Dokumenttyp: Journalartikel / Zeitschriftenartikel
Abstract: Microglia are known as the most motile cells in the central nervous system (CNS). It was shown in vivo that they permanently scan their direct microenvironment and react to pathological conditions within minutes. Many studies of brain pathologies use fixed brain tissue to investigate cellular changes. Unfortunately, due to technical reasons, the time span between the induction of the fixation procedure (start of the perfusion) and the finally-fixed tissue lasts several minutes, giving time to microglia to start reacting to the ischemic conditions due to perfusion start. Here, we investigated the microglial changes generated by the fixation itself in TgH(CX3CR1-EGFP) mice with fluorescent labelled microglia using confocal laser scanning microscopy (CLSM) of fixed brain tissue as well as two-photon laser scanning microscopy (2P-LSM) during the perfusion of a living animal. We revealed the impact of fixation and buffer parameters on cell morphology. The largest morphological differences compared to physiological in vivo branch arborization were observed when the directly dissected brain was immersed in paraformaldehyde fixation solution overnight, without prior fixative perfusion of the animal. But even perfusion with a fixative, followed by post-fixation leads to small changes in microglial process length and number and could not be prevented when compared to physiological in vivo microglia morphology acquired using in vivo 2P-LSM. Interestingly, perfusion with different buffers either oxygenated artificial cerebrospinal fluid or phosphate buffered saline prior to perfusion-fixation showed minor microglia changes in arborization and/or number of processes. Fixation methods influence microglia morphology. Therefore, to define microglia activation states immunohistochemical stainings or genetic labelling of the cells have to be included in addition to morphological analysis.
DOI der Erstveröffentlichung: 10.1016/j.jchemneu.2017.08.007
URL der Erstveröffentlichung: https://www.sciencedirect.com/science/article/abs/pii/S0891061817300571
Link zu diesem Datensatz: urn:nbn:de:bsz:291--ds-395666
hdl:20.500.11880/35660
http://dx.doi.org/10.22028/D291-39566
ISSN: 0891-0618
Datum des Eintrags: 18-Apr-2023
Fakultät: M - Medizinische Fakultät
Fachrichtung: M - Physiologie
Professur: M - Prof. Dr. Frank Kirchhoff
Sammlung:SciDok - Der Wissenschaftsserver der Universität des Saarlandes

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