From fix to fit into the autoptic human brains

Beatrice Paradiso, M Simonato, Gaetano Thiene, Anna M. Lavezzi

Research output: Contribution to journalArticle

Abstract

Formalin-fixed, paraffinembedded (FFPE) human brain tissues are very often stored in formalin for long time. Formalin fixation reduces immunostaining, and the DNA/RNA extraction from FFPE brain tissue becomes suboptimal. At present, there are different protocols of fixation and several procedures and kits to extract DNA/RNA from paraffin embedding tissue, but a gold standard protocol remains distant. In this study, we analyzed four types of fixation systems and compared histo and immuno-staining. Based on our results, we propose a modified method of combined fixation in formalin and formic acid for the autoptic adult brain to obtain easy, fast, safe and efficient immunolabelling of long-stored FFPE tissue. In particular, we have achieved an improved preservation of cellular morphology and obtained success in postmortem immunostaining for NeuN. This nuclear antigen is an important marker for mapping neurons, for example, to evaluate the histopathology of temporal lobe epilepsy or to draw the topography of cardiorespiratory brainstem nuclei in sudden infant death syndrome (SIDS). However, NeuN staining is frequently faint or lost in postmortem human brain tissues. In addition, we attained Fluoro Jade C staining, a marker of neurodegeneration, and immunofluorescent staining for stem cell antigens in the postnatal human brain, utilizing custom fit fixation procedures.
Original languageEnglish
Article number2944
JournalEuropean journal of histochemistry : EJH
Volume62
Issue number3
DOIs
Publication statusPublished - 2018

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Formaldehyde
Brain
Staining and Labeling
formic acid
Paraffin Embedding
RNA
Nuclear Antigens
Sudden Infant Death
Temporal Lobe Epilepsy
DNA
Brain Stem
Stem Cells
Neurons
Antigens

Cite this

Paradiso, B., Simonato, M., Thiene, G., & Lavezzi, A. M. (2018). From fix to fit into the autoptic human brains. European journal of histochemistry : EJH, 62(3), [2944]. https://doi.org/10.4081/ejh.2018.2944

From fix to fit into the autoptic human brains. / Paradiso, Beatrice; Simonato, M; Thiene, Gaetano; Lavezzi, Anna M.

In: European journal of histochemistry : EJH, Vol. 62, No. 3, 2944, 2018.

Research output: Contribution to journalArticle

Paradiso, B, Simonato, M, Thiene, G & Lavezzi, AM 2018, 'From fix to fit into the autoptic human brains', European journal of histochemistry : EJH, vol. 62, no. 3, 2944. https://doi.org/10.4081/ejh.2018.2944
Paradiso, Beatrice ; Simonato, M ; Thiene, Gaetano ; Lavezzi, Anna M. / From fix to fit into the autoptic human brains. In: European journal of histochemistry : EJH. 2018 ; Vol. 62, No. 3.
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