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000165344 037__ $$aDZNE-2022-01622
000165344 041__ $$aEnglish
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000165344 1001_ $$aPérez Arévalo, Andrea$$b0
000165344 245__ $$aTrans-cardiac perfusion of neonatal mice and immunofluorescence of the whole body as a method to study nervous system development.
000165344 260__ $$aSan Francisco, California, US$$bPLOS$$c2022
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000165344 520__ $$aWhole animal perfusion is a well-established method that has been used for the past decades in multiple research fields. Particularly, it has been very important for the study of the brain. The rapid and uniform fixation of tissue is essential for the preservation of its integrity and the study of complex structures. For small tissue pieces submerging in formaldehyde solution oftentimes is sufficient to get a good fixation, larger tissues or organs with a more complicated structure present a greater difficulty. Here, we report the precise parameters to successfully perform trans-cardiac perfusion of neonatal mouse pups that allows a uniform fixation of the whole body for subsequent structural analysis and immunohistochemistry. In comparison to standard perfusion procedures of adult mice, changes in the pump velocity, the buffer volume and in the needle size lead to high quality fixation of neonatal mice pups. Further, we present a whole-body section staining, which results in a highly specific immunofluorescence signal suited for detailed analysis of multiple tissues or systems at the same time. Thus, our protocol provides a reproducible and reliable method for neonatal perfusion and staining that can rapidly be applied in any laboratory. It allows a high quality analysis of cellular structures and expression profiles at early developmental stages.
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000165344 650_7 $$01HG84L3525$$2NLM Chemicals$$aFormaldehyde
000165344 650_2 $$2MeSH$$aAnimals
000165344 650_2 $$2MeSH$$aAnimals, Newborn
000165344 650_2 $$2MeSH$$aBrain
000165344 650_2 $$2MeSH$$aFluorescent Antibody Technique
000165344 650_2 $$2MeSH$$aFormaldehyde
000165344 650_2 $$2MeSH$$aImmunohistochemistry
000165344 650_2 $$2MeSH$$aMice
000165344 650_2 $$2MeSH$$aPerfusion: methods
000165344 650_2 $$2MeSH$$aStaining and Labeling
000165344 650_2 $$2MeSH$$aTissue Fixation: methods
000165344 7001_ $$00000-0002-3972-2057$$aLutz, Anne-Kathrin$$b1
000165344 7001_ $$aAtanasova, Ekaterina$$b2
000165344 7001_ $$0P:(DE-2719)2812855$$aBöckers, Tobias$$b3$$eLast author$$udzne
000165344 773__ $$0PERI:(DE-600)2267670-3$$a10.1371/journal.pone.0275780$$gVol. 17, no. 10, p. e0275780 -$$n10$$pe0275780$$tPLOS ONE$$v17$$x1932-6203$$y2022
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