title: The workflow from post-mortem human brain sampling to cell microdissection: a Brain Net Europe study.
abstract: Brain banks manage and store fully clinically and pathologically characterised brains. The diversity of techniques used in research projects increases. These biological resource centres are made to adapt brain tissue processing. Furthermore, the development of more sensitive techniques to analyse nucleic acids and proteins offers new fields of exploration when combined with laser capture microdissection in order to decipher the physiopathology of diseases at the cell level. In this study, our goal was to evaluate procedures and set a workflow compatible with the constraints of brain banks, from brain sampling to laser capture microdissection and pre-analytical quality assessment. We compared various methods of freezing brain tissue, focused on morphological quality preservation of brain microscopical structures and on the quality of nucleic acid or protein yields. Staining protocols combined with strategies to lower neurones autofluorescence were adapted for the same purpose. Finally, we found that laser capture microdissection is possible in the setting of brain banks. However, the entire process has to be envisioned from the autopsy to the analysis. The impact on protein or nucleic acid quality is a limitation that restricts the amount of samples available for this purpose.
authors.affiliation: INSERM U1028; CNRS UM5292; Lyon Neuroscience Research Center, Neuro-oncology and Neuro-inflammation Team, 69000, Lyon, France, david.meyronet@chu-lyon.fr.
authors.forename: David
authors.initials: D
authors.lastname: Meyronet
authors.forename: Aline
authors.initials: A
authors.lastname: Dorey
authors.forename: Patrick
authors.initials: P
authors.lastname: Massoma
authors.forename: Catherine
authors.initials: C
authors.lastname: Rey
authors.forename: Eudeline
authors.initials: E
authors.lastname: Alix
authors.forename: Karen
authors.initials: K
authors.lastname: Silva
authors.forename: Corinne
authors.initials: C
authors.lastname: Perrin
authors.forename: Isabelle
authors.initials: I
authors.lastname: Quadrio
authors.forename: Armand
authors.initials: A
authors.lastname: Perret-Liaudet
authors.forename: Nathalie
authors.initials: N
authors.lastname: Streichenberger
authors.forename: Nicole
authors.initials: N
authors.lastname: Thomasset
authors.forename: Jérôme
authors.initials: J
authors.lastname: Honnorat
authors.forename: Thomas
authors.initials: T
authors.lastname: Arzberger
authors.forename: Hans
authors.initials: H
authors.lastname: Kretzschmar
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D001921:Brain / Q000473:pathology*; D001927:Brain Diseases / Q000473:pathology; D005060:Europe; D005260:Female; D006801:Humans; D008297:Male; D042282:Microdissection; D008875:Middle Aged; D009474:Neurons / Q000378:metabolism / Q000473:pathology*; D011180:Postmortem Changes; D011506:Proteins / Q000235:genetics / Q000378:metabolism; D013048:Specimen Handling; D013194:Staining and Labeling; D014015:Tissue Banks; D057188:Workflow
chemical_list: D011506:Proteins