title: Preparation of human formalin-fixed brain slices for electron microscopic investigations.
abstract: Ultra-structural analysis of human post-mortem brain tissue is important for investigations into the pathomechanism of neuropsychiatric disorders, especially those lacking alternative models of studying human-specific morphological features. For example, Von Economo Neurons (VENs) mainly located in the anterior cingulate cortex and in the anterior part of the insula, which seem to play a role in a variety of neuropsychiatric conditions, including frontotemporal dementia, autism and schizophrenia, can hardly be studied in nonhuman animals. Accordingly, little is known about the ultra-structural alterations of these neurons, though important research using qualitative stereological methods has revealed that protein expression of the VENs assigns them a role in immune function. Formaldehyde, which is the most common fixative in human pathology, interferes with the immunoreactivity of the tissue, possibly leading to unreliable results. Therefore, a method for ultra-structural investigations independent of antigenic properties of the fixated tissue is needed. Here, we propose an approach using electron microscopy to examine cytoskeletal structures, synapses and mitochondria in these cells. We also show that our methodology is able to keep tissue consumption to a minimum, while still allowing for the specimens to be handled with ease by using agar embedded slices in contrast to blocks for the embedding procedure. Accordingly, a stepwise protocol utilising 60μm thick human post mortem brain sections for electron microscopic ultra-structural investigations is presented.
authors.affiliation: Institute of Anatomy, Department of Cytology, Ruhr-University Bochum, Germany.
authors.forename: Martin
authors.initials: M
authors.lastname: Krause
authors.affiliation: LWL University Hospital Bochum, Division of Cognitive Neuropsychiatry and Psychiatric Preventive Medicine, Ruhr-University Bochum, Germany.
authors.forename: Martin
authors.initials: M
authors.lastname: Brüne
authors.affiliation: Institute of Anatomy, Department of Cytology, Ruhr-University Bochum, Germany. Electronic address: carsten.theiss@rub.de.
authors.forename: Carsten
authors.initials: C
authors.lastname: Theiss
mesh_terms: D000362:Agar / Q000737:chemistry; D002102:Cadaver; D005404:Fixatives / Q000737:chemistry; D005557:Formaldehyde / Q000737:chemistry*; D006179:Gyrus Cinguli / Q000737:chemistry* / Q000473:pathology / Q000648:ultrastructure*; D006801:Humans; D008854:Microscopy, Electron / Q000379:methods*; D009474:Neurons / Q000737:chemistry* / Q000473:pathology / Q000648:ultrastructure*; D010301:Parkinson Disease, Postencephalitic / Q000473:pathology; D013048:Specimen Handling / Q000379:methods; D016610:Tissue Embedding / Q000379:methods
chemical_list: D005404:Fixatives; D005557:Formaldehyde; D000362:Agar
keywords: Electron microscopy; Human formalin fixed brain slices; Post mortem tissue; Schizophrenia; Ultrastructure, Von Economo Neuron