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@ARTICLE{PrezArvalo:165344,
author = {Pérez Arévalo, Andrea and Lutz, Anne-Kathrin and
Atanasova, Ekaterina and Böckers, Tobias},
title = {{T}rans-cardiac perfusion of neonatal mice and
immunofluorescence of the whole body as a method to study
nervous system development.},
journal = {PLOS ONE},
volume = {17},
number = {10},
issn = {1932-6203},
address = {San Francisco, California, US},
publisher = {PLOS},
reportid = {DZNE-2022-01622},
pages = {e0275780},
year = {2022},
abstract = {Whole animal perfusion is a well-established method that
has been used for the past decades in multiple research
fields. Particularly, it has been very important for the
study of the brain. The rapid and uniform fixation of tissue
is essential for the preservation of its integrity and the
study of complex structures. For small tissue pieces
submerging in formaldehyde solution oftentimes is sufficient
to get a good fixation, larger tissues or organs with a more
complicated structure present a greater difficulty. Here, we
report the precise parameters to successfully perform
trans-cardiac perfusion of neonatal mouse pups that allows a
uniform fixation of the whole body for subsequent structural
analysis and immunohistochemistry. In comparison to standard
perfusion procedures of adult mice, changes in the pump
velocity, the buffer volume and in the needle size lead to
high quality fixation of neonatal mice pups. Further, we
present a whole-body section staining, which results in a
highly specific immunofluorescence signal suited for
detailed analysis of multiple tissues or systems at the same
time. Thus, our protocol provides a reproducible and
reliable method for neonatal perfusion and staining that can
rapidly be applied in any laboratory. It allows a high
quality analysis of cellular structures and expression
profiles at early developmental stages.},
keywords = {Animals / Animals, Newborn / Brain / Fluorescent Antibody
Technique / Formaldehyde / Immunohistochemistry / Mice /
Perfusion: methods / Staining and Labeling / Tissue
Fixation: methods / Formaldehyde (NLM Chemicals)},
cin = {AG Böckers},
ddc = {610},
cid = {I:(DE-2719)1910002},
pnm = {352 - Disease Mechanisms (POF4-352)},
pid = {G:(DE-HGF)POF4-352},
typ = {PUB:(DE-HGF)16},
pubmed = {pmid:36227942},
pmc = {pmc:PMC9560478},
doi = {10.1371/journal.pone.0275780},
url = {https://pub.dzne.de/record/165344},
}