gms | German Medical Science

Joint-Meeting of the German Society for Neuropathology and Neuroanatomy (DGNN) and the Scandinavian Neuropathological Society (SNS)

Deutsche Gesellschaft für Neuropathologie und Neuroanatomie

22.09.-24.09.2016, Hamburg

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Label-free multiphoton microscopy extends the neuropathological armamentarium: Visualization of hippocampal sclerosis

Meeting Abstract

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  • presenting/speaker Ortrud Uckermann - Universitätsklinikum Dresden, Klinik und Poliklinik für Neurochirurgie, Dresden, Germany
  • Roberta Galli - TU Dresden, Klinisches Sensoring und Monitoring, Klinik und Poliklinik für Anästhesiologie und Intensivtherapie, Dresden, Germany
  • Roland Coras - Universitätsklinikum Erlangen, Neuropathologisches Institut, Erlangen, Germany
  • Matthias Meinhardt - Universitätsklinikum Dresden, Institut für Pathologie, Dresden, Germany
  • Ingmar Blümcke - Universitätsklinikum Erlangen, Neuropathologisches Institut, Erlangen, Germany
  • Gerald Steiner - TU Dresden, Klinisches Sensoring und Monitoring, Klinik und Poliklinik für Anästhesiologie und Intensivtherapie, Dresden, Germany
  • Matthias Kirsch - Universitätsklinikum Dresden, Klinik und Poliklinik für Neurochirurgie, Dresden, Germany

Deutsche Gesellschaft für Neuropathologie und Neuroanatomie. Scandinavian Neuropathological Society. Joint-Meeting of the German Society for Neuropathology and Neuroanatomy (DGNN) and the Scandinavian Neuropathological Society (SNS). Hamburg, 22.-24.09.2016. Düsseldorf: German Medical Science GMS Publishing House; 2016. Doc16dgnnP04

doi: 10.3205/16dgnn17, urn:nbn:de:0183-16dgnn172

Veröffentlicht: 14. September 2016

© 2016 Uckermann et al.
Dieser Artikel ist ein Open-Access-Artikel und steht unter den Lizenzbedingungen der Creative Commons Attribution 4.0 License (Namensnennung). Lizenz-Angaben siehe http://creativecommons.org/licenses/by/4.0/.

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Question: Multiphoton microscopy can be applied to fresh tissue and visualizes morphochemical properties and structure without the need for labeling or tissue preparation. Coherent anti-Stokes Raman scattering (CARS) addresses the lipid content, second harmonic generation (SHG) shows collagen, and two-photon excited fluorescence (TPEF) visualizes endogenous fluorophores of the tissue. We investigated this technique to detect pathological changes in hippocampal tissue of patients with pharmacoresistant temporo-mesial epilepsy.

Methods: Samples of human hippocampal tissue (n=15) were obtained during routine surgeries. CARS, SHG and TPEF were acquired simultaneously on unstained cryosections and fresh unfixed samples. Reference histopathology was performed.

Results: Multiphoton microscopy revealed hippocampal layering and micromorphological details: CARS displayed white and gray matter layers and allowed the assessment of axonal myelin. SHG revealed the size and location of blood vessels based on adventitial collagen. Additionally, corpora amylacea (CoA) were found to be SHG-active. They were predominantly located close to blood vessels and in areas with marked neuronal loss. Pyramidal cell bodies were characterized by intense cytoplasmatic endogenous TPEF, while the fluorescence of granular and non-neuronal cells was less pronounced. The lack of TPEF-positive pyramidal neurons in combination with massive deposition of CoA allowed the identification of hippocampus sclerosis (HS).

Conclusion: We present a label-free and fast optical approach that analyses pathological aspects of HS. Hippocampal layering, loss of pyramidal cells and presence of CoA indicative of sclerosis are easily identifiable.In conclusion, label-free multiphoton microscopy has the potential to extend the histopathological armamentarium for assessment of changes of the hippocampal formation.