TY  - JOUR
AU  - Pérez Arévalo, Andrea
AU  - Lutz, Anne-Kathrin
AU  - Atanasova, Ekaterina
AU  - Böckers, Tobias
TI  - Trans-cardiac perfusion of neonatal mice and immunofluorescence of the whole body as a method to study nervous system development.
JO  - PLOS ONE
VL  - 17
IS  - 10
SN  - 1932-6203
CY  - San Francisco, California, US
PB  - PLOS
M1  - DZNE-2022-01622
SP  - e0275780
PY  - 2022
AB  - 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.
KW  - Animals
KW  - Animals, Newborn
KW  - Brain
KW  - Fluorescent Antibody Technique
KW  - Formaldehyde
KW  - Immunohistochemistry
KW  - Mice
KW  - Perfusion: methods
KW  - Staining and Labeling
KW  - Tissue Fixation: methods
KW  - Formaldehyde (NLM Chemicals)
LB  - PUB:(DE-HGF)16
C6  - pmid:36227942
C2  - pmc:PMC9560478
DO  - DOI:10.1371/journal.pone.0275780
UR  - https://pub.dzne.de/record/165344
ER  -