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The workflow from post-mortem human brain sampling to cell microdissection: a Brain Net Europe study

  • Translational Neurosciences - Original Article
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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.

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Acknowledgments

We thank Rachel Plantier and Marie Pierre Hoba for their skilful technical assistance. This study was supported by the European Union grant FP6: BNEII No LSHM-CT-2004-503039 and Centre Mémoire Ressources de Lyon (Dr. Bernard Croisile, Pr Pierre Krolak-Salmon, Dr. Marie-Hélène Coste). Frozen samples used in this study were provided by Hospices Civils de Lyon Brain Bank, Cardiobiotec, Centre de ressources biologies des Hospices Civils de Lyon.

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Authors and Affiliations

  1. INSERM U1028; CNRS UM5292; Lyon Neuroscience Research Center, Neuro-oncology and Neuro-inflammation Team, 69000, Lyon, France

    David Meyronet, Patrick Massoma, Nicole Thomasset & Jérôme Honnorat

  2. “ProfileXpert”, service de “Génomique & Microgénomique”, Université Lyon 1, SFR santé LYON-EST, UCBL-INSERM US 7-CNRS UMS 3453, 8 av. Rockefeller, 69373, Lyon Cedex 08, France

    Catherine Rey

  3. Neurochemistry Unit, Department of Biochemistry, Hospices Civils de Lyon, Groupement Hospitalier Est, Lyon1 University, Lyon Neuroscience Research Center, INSERM, Lyon, France

    Isabelle Quadrio & Armand Perret-Liaudet

  4. Hospices Civils de Lyon, Centre de Pathologie et de Neuropathologie Est, 69003, Lyon, France

    David Meyronet, Aline Dorey, Eudeline Alix, Karen Silva, Corinne Perrin & Nathalie Streichenberger

  5. INSERM U1028; CNRS UM5292; Lyon Neuroscience Research Center, BIORAN Team, 69000, Lyon, France

    Isabelle Quadrio, Armand Perret-Liaudet & Nathalie Streichenberger

  6. Hospices Civils de Lyon, Service de Neurologie, 69003, Lyon, France

    Jérôme Honnorat

  7. Centre for Neuropathology and Prion Research, Ludwig-Maximilians-University, Munich, Germany

    Thomas Arzberger & Hans Kretzschmar

  8. Department of Psychiatry and Psychotherapy, Ludwig-Maximilians-University, Munich, Germany

    Thomas Arzberger

  9. Centre de Pathologie et Neuropathologie Est, Hospices Civils de Lyon, Groupement Hospitalier Est, Bat A3, 6ème étage, 57 Bd Pinel, 69667, Bron, France

    David Meyronet

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  1. David Meyronet
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  2. Aline Dorey
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  3. Patrick Massoma
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  4. Catherine Rey
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  9. Armand Perret-Liaudet
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  10. Nathalie Streichenberger
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  11. Nicole Thomasset
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  13. Thomas Arzberger
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  14. Hans Kretzschmar
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Correspondence to David Meyronet.

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Meyronet, D., Dorey, A., Massoma, P. et al. The workflow from post-mortem human brain sampling to cell microdissection: a Brain Net Europe study. J Neural Transm 122, 975–991 (2015). https://doi.org/10.1007/s00702-015-1378-4

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