Skip to main content
Top
Published in: Acta Neurochirurgica 6/2023

Open Access 06-05-2023 | Epilepsy | Technical Note

Technical report: surgical preparation of human brain tissue for clinical and basic research

Authors: J. Straehle, V. M. Ravi, D.H. Heiland, C. Galanis, M. Lenz, Junyi Zhang, N.N Neidert, A. El Rahal, I. Vasilikos, P. Kellmeyer, C. Scheiwe, J.H. Klingler, C. Fung, A. Vlachos, J. Beck, O. Schnell

Published in: Acta Neurochirurgica | Issue 6/2023

Login to get access

Abstract

Background

The study of the distinct structure and function of the human central nervous system, both in healthy and diseased states, is becoming increasingly significant in the field of neuroscience. Typically, cortical and subcortical tissue is discarded during surgeries for tumors and epilepsy. Yet, there is a strong encouragement to utilize this tissue for clinical and basic research in humans. Here, we describe the technical aspects of the microdissection and immediate handling of viable human cortical access tissue for basic and clinical research, highlighting the measures needed to be taken in the operating room to ensure standardized procedures and optimal experimental results.

Methods

In multiple rounds of experiments (n = 36), we developed and refined surgical principles for the removal of cortical access tissue. The specimens were immediately immersed in cold carbogenated N-methyl-D-glucamine-based artificial cerebrospinal fluid for electrophysiology and electron microscopy experiments or specialized hibernation medium for organotypic slice cultures.

Results

The surgical principles of brain tissue microdissection were (1) rapid preparation (<1 min), (2) maintenance of the cortical axis, (3) minimization of mechanical trauma to sample, (4) use of pointed scalpel blade, (5) avoidance of cauterization and blunt preparation, (6) constant irrigation, and (7) retrieval of the sample without the use of forceps or suction. After a single round of introduction to these principles, multiple surgeons adopted the technique for samples with a minimal dimension of 5 mm spanning all cortical layers and subcortical white matter. Small samples (5–7 mm) were ideal for acute slice preparation and electrophysiology. No adverse events from sample resection were observed.

Conclusion

The microdissection technique of human cortical access tissue is safe and easily adoptable into the routine of neurosurgical procedures. The standardized and reliable surgical extraction of human brain tissue lays the foundation for human-to-human translational research on human brain tissue.
Appendix
Available only for authorised users
Literature
2.
go back to reference Campagnola L, Seeman SC, Chartrand T, Kim L, Hoggarth A, Gamlin C, Ito S, Trinh J, Davoudian P, Radaelli C, Kim M-H, Hage T, Braun T, Alfiler L, Andrade J, Bohn P, Dalley R, Henry A, Kebede S et al Local connectivity and synaptic dynamics in mouse and human neocortex. Science 375:eabj5861. https://doi.org/10.1126/science.abj5861 Campagnola L, Seeman SC, Chartrand T, Kim L, Hoggarth A, Gamlin C, Ito S, Trinh J, Davoudian P, Radaelli C, Kim M-H, Hage T, Braun T, Alfiler L, Andrade J, Bohn P, Dalley R, Henry A, Kebede S et al Local connectivity and synaptic dynamics in mouse and human neocortex. Science 375:eabj5861. https://​doi.​org/​10.​1126/​science.​abj5861
4.
go back to reference Duffau H (2020) Functional mapping before and after low-grade glioma surgery: a new way to decipher various spatiotemporal patterns of individual neuroplastic potential in brain tumor patients. Cancers 12:2611CrossRefPubMedPubMedCentral Duffau H (2020) Functional mapping before and after low-grade glioma surgery: a new way to decipher various spatiotemporal patterns of individual neuroplastic potential in brain tumor patients. Cancers 12:2611CrossRefPubMedPubMedCentral
5.
go back to reference Fiala JC, Spacek J, Harris KM (2002) Dendritic spine pathology: cause or consequence of neurological disorders? Brain Res Rev 39:29–54CrossRefPubMed Fiala JC, Spacek J, Harris KM (2002) Dendritic spine pathology: cause or consequence of neurological disorders? Brain Res Rev 39:29–54CrossRefPubMed
7.
go back to reference Henrik Heiland D, Ravi VM, Behringer SP, Frenking JH, Wurm J, Joseph K, Garrelfs NWC, Strähle J, Heynckes S, Grauvogel J, Franco P, Mader I, Schneider M, Potthoff A-L, Delev D, Hofmann UG, Fung C, Beck J, Sankowski R et al (2019) Tumor-associated reactive astrocytes aid the evolution of immunosuppressive environment in glioblastoma. Nat Commun 10:2541. https://doi.org/10.1038/s41467-019-10493-6CrossRefPubMedPubMedCentral Henrik Heiland D, Ravi VM, Behringer SP, Frenking JH, Wurm J, Joseph K, Garrelfs NWC, Strähle J, Heynckes S, Grauvogel J, Franco P, Mader I, Schneider M, Potthoff A-L, Delev D, Hofmann UG, Fung C, Beck J, Sankowski R et al (2019) Tumor-associated reactive astrocytes aid the evolution of immunosuppressive environment in glioblastoma. Nat Commun 10:2541. https://​doi.​org/​10.​1038/​s41467-019-10493-6CrossRefPubMedPubMedCentral
8.
go back to reference Hollon TC, Pandian B, Adapa AR, Urias E, Save AV, Khalsa SSS, Eichberg DG, D’Amico RS, Farooq ZU, Lewis S, Petridis PD, Marie T, Shah AH, Garton HJL, Maher CO, Heth JA, McKean EL, Sullivan SE, Hervey-Jumper SL et al (2020) Near real-time intraoperative brain tumor diagnosis using stimulated Raman histology and deep neural networks. Nat Med 26:52–58. https://doi.org/10.1038/s41591-019-0715-9CrossRefPubMedPubMedCentral Hollon TC, Pandian B, Adapa AR, Urias E, Save AV, Khalsa SSS, Eichberg DG, D’Amico RS, Farooq ZU, Lewis S, Petridis PD, Marie T, Shah AH, Garton HJL, Maher CO, Heth JA, McKean EL, Sullivan SE, Hervey-Jumper SL et al (2020) Near real-time intraoperative brain tumor diagnosis using stimulated Raman histology and deep neural networks. Nat Med 26:52–58. https://​doi.​org/​10.​1038/​s41591-019-0715-9CrossRefPubMedPubMedCentral
10.
go back to reference Kato H, Ito Z, Matsuoka S, Sakurai Y (1973) Electrical activities of neurons in the sliced human cortex in vitro. Electroencephalogr Clin Neurophysiol 35:457–462CrossRefPubMed Kato H, Ito Z, Matsuoka S, Sakurai Y (1973) Electrical activities of neurons in the sliced human cortex in vitro. Electroencephalogr Clin Neurophysiol 35:457–462CrossRefPubMed
12.
go back to reference Kovacs FE, Knop T, Urbanski MJ, Freiman I, Freiman TM, Feuerstein TJ, Zentner J, Szabo B (2012) Exogenous and endogenous cannabinoids suppress inhibitory neurotransmission in the human neocortex. Neuropsychopharmacology 37:1104–1114CrossRefPubMed Kovacs FE, Knop T, Urbanski MJ, Freiman I, Freiman TM, Feuerstein TJ, Zentner J, Szabo B (2012) Exogenous and endogenous cannabinoids suppress inhibitory neurotransmission in the human neocortex. Neuropsychopharmacology 37:1104–1114CrossRefPubMed
14.
go back to reference Le Duigou C, Savary E, Morin-Brureau M, Gomez-Dominguez D, Sobczyk A, Chali F, Milior G, Kraus L, Meier JC, Kullmann DM (2018) Imaging pathological activities of human brain tissue in organotypic culture. J Neurosci Methods 298:33–44CrossRefPubMedPubMedCentral Le Duigou C, Savary E, Morin-Brureau M, Gomez-Dominguez D, Sobczyk A, Chali F, Milior G, Kraus L, Meier JC, Kullmann DM (2018) Imaging pathological activities of human brain tissue in organotypic culture. J Neurosci Methods 298:33–44CrossRefPubMedPubMedCentral
15.
go back to reference Lenz M, Kruse P, Eichler A, Straehle J, Beck J, Deller T, Vlachos A (2021) All-trans retinoic acid induces synaptic plasticity in human cortical neurons. Elife 10:e63026CrossRefPubMedPubMedCentral Lenz M, Kruse P, Eichler A, Straehle J, Beck J, Deller T, Vlachos A (2021) All-trans retinoic acid induces synaptic plasticity in human cortical neurons. Elife 10:e63026CrossRefPubMedPubMedCentral
17.
go back to reference Maier JP, Ravi VM, Kueckelhaus J, Behringer SP, Garrelfs N, Will P, Sun N, von Ehr J, Goeldner JM, Pfeifer D, Follo M, Hannibal L, Walch AK, Hofmann UG, Beck J, Heiland DH, Schnell O, Joseph K (2021) Inhibition of metabotropic glutamate receptor III facilitates sensitization to alkylating chemotherapeutics in glioblastoma. Cell Death Dis 12:723–723. https://doi.org/10.1038/s41419-021-03937-9CrossRefPubMedPubMedCentral Maier JP, Ravi VM, Kueckelhaus J, Behringer SP, Garrelfs N, Will P, Sun N, von Ehr J, Goeldner JM, Pfeifer D, Follo M, Hannibal L, Walch AK, Hofmann UG, Beck J, Heiland DH, Schnell O, Joseph K (2021) Inhibition of metabotropic glutamate receptor III facilitates sensitization to alkylating chemotherapeutics in glioblastoma. Cell Death Dis 12:723–723. https://​doi.​org/​10.​1038/​s41419-021-03937-9CrossRefPubMedPubMedCentral
18.
go back to reference Mohan H, Verhoog MB, Doreswamy KK, Eyal G, Aardse R, Lodder BN, Goriounova NA, Asamoah B, Brakspear ABC, Groot C, van der Sluis S, Testa-Silva G, Obermayer J, ZSRM B, Narayanan RT, Baayen JC, Segev I, Mansvelder HD, de Kock CPJ (2015) Dendritic and axonal architecture of individual pyramidal neurons across layers of adult human neocortex. Cereb Cortex 25:4839–4853. https://doi.org/10.1093/cercor/bhv188CrossRefPubMedPubMedCentral Mohan H, Verhoog MB, Doreswamy KK, Eyal G, Aardse R, Lodder BN, Goriounova NA, Asamoah B, Brakspear ABC, Groot C, van der Sluis S, Testa-Silva G, Obermayer J, ZSRM B, Narayanan RT, Baayen JC, Segev I, Mansvelder HD, de Kock CPJ (2015) Dendritic and axonal architecture of individual pyramidal neurons across layers of adult human neocortex. Cereb Cortex 25:4839–4853. https://​doi.​org/​10.​1093/​cercor/​bhv188CrossRefPubMedPubMedCentral
20.
go back to reference Narayanan RT, Mohan H, Broersen R, de Haan R, Pieneman AW, de Kock CP (2014) Juxtasomal biocytin labeling to study the structure-function relationship of individual cortical neurons. J Vis Exp 84:e51359 Narayanan RT, Mohan H, Broersen R, de Haan R, Pieneman AW, de Kock CP (2014) Juxtasomal biocytin labeling to study the structure-function relationship of individual cortical neurons. J Vis Exp 84:e51359
21.
go back to reference Neidert N, Straehle J, Erny D, Sacalean V, El Rahal A, Steybe D, Schmelzeisen R, Vlachos A, Reinacher PC, Coenen VA, Mizaikoff B, Heiland DH, Prinz M, Beck J, Schnell O (2022) Stimulated Raman histology in the neurosurgical workflow of a major European neurosurgical center — part A. Neurosurg Rev 45:1731–1739. https://doi.org/10.1007/s10143-021-01712-0CrossRefPubMed Neidert N, Straehle J, Erny D, Sacalean V, El Rahal A, Steybe D, Schmelzeisen R, Vlachos A, Reinacher PC, Coenen VA, Mizaikoff B, Heiland DH, Prinz M, Beck J, Schnell O (2022) Stimulated Raman histology in the neurosurgical workflow of a major European neurosurgical center — part A. Neurosurg Rev 45:1731–1739. https://​doi.​org/​10.​1007/​s10143-021-01712-0CrossRefPubMed
22.
go back to reference Orringer DA, Pandian B, Niknafs YS, Hollon TC, Boyle J, Lewis S, Garrard M, Hervey-Jumper SL, Garton HJL, Maher CO, Heth JA, Sagher O, Wilkinson DA, Snuderl M, Venneti S, Ramkissoon SH, McFadden KA, Fisher-Hubbard A, Lieberman AP et al (2017) Rapid intraoperative histology of unprocessed surgical specimens via fibre-laser-based stimulated Raman scattering microscopy. Nat Biomed Eng 1:0027. https://doi.org/10.1038/s41551-016-0027CrossRefPubMedPubMedCentral Orringer DA, Pandian B, Niknafs YS, Hollon TC, Boyle J, Lewis S, Garrard M, Hervey-Jumper SL, Garton HJL, Maher CO, Heth JA, Sagher O, Wilkinson DA, Snuderl M, Venneti S, Ramkissoon SH, McFadden KA, Fisher-Hubbard A, Lieberman AP et al (2017) Rapid intraoperative histology of unprocessed surgical specimens via fibre-laser-based stimulated Raman scattering microscopy. Nat Biomed Eng 1:0027. https://​doi.​org/​10.​1038/​s41551-016-0027CrossRefPubMedPubMedCentral
23.
24.
go back to reference Ravi VM, Neidert N, Will P, Joseph K, Maier JP, Kückelhaus J, Vollmer L, Goeldner JM, Behringer SP, Scherer F, Boerries M, Follo M, Weiss T, Delev D, Kernbach J, Franco P, Schallner N, Dierks C, Carro MS et al (2022a) T-cell dysfunction in the glioblastoma microenvironment is mediated by myeloid cells releasing interleukin-10. Nat Commun 13:925. https://doi.org/10.1038/s41467-022-28523-1CrossRefPubMedPubMedCentral Ravi VM, Neidert N, Will P, Joseph K, Maier JP, Kückelhaus J, Vollmer L, Goeldner JM, Behringer SP, Scherer F, Boerries M, Follo M, Weiss T, Delev D, Kernbach J, Franco P, Schallner N, Dierks C, Carro MS et al (2022a) T-cell dysfunction in the glioblastoma microenvironment is mediated by myeloid cells releasing interleukin-10. Nat Commun 13:925. https://​doi.​org/​10.​1038/​s41467-022-28523-1CrossRefPubMedPubMedCentral
25.
go back to reference Ravi VM, Will P, Kueckelhaus J, Sun N, Joseph K, Salié H, Vollmer L, Kuliesiute U, von Ehr J, Benotmane JK, Neidert N, Follo M, Scherer F, Goeldner JM, Behringer SP, Franco P, Khiat M, Zhang J, Hofmann UG et al (2022b) Spatially resolved multi-omics deciphers bidirectional tumor-host interdependence in glioblastoma. Cancer Cell 40:639–655.e13. https://doi.org/10.1016/j.ccell.2022.05.009CrossRefPubMed Ravi VM, Will P, Kueckelhaus J, Sun N, Joseph K, Salié H, Vollmer L, Kuliesiute U, von Ehr J, Benotmane JK, Neidert N, Follo M, Scherer F, Goeldner JM, Behringer SP, Franco P, Khiat M, Zhang J, Hofmann UG et al (2022b) Spatially resolved multi-omics deciphers bidirectional tumor-host interdependence in glioblastoma. Cancer Cell 40:639–655.e13. https://​doi.​org/​10.​1016/​j.​ccell.​2022.​05.​009CrossRefPubMed
26.
go back to reference Reinecke D, von Spreckelsen N, Mawrin C, Ion-Margineanu A, Fürtjes G, Jünger ST, Khalid F, Freudiger CW, Timmer M, Ruge MI, Goldbrunner R, Neuschmelting V (2022) Novel rapid intraoperative qualitative tumor detection by a residual convolutional neural network using label-free stimulated Raman scattering microscopy. Acta Neuropathol Commun 10:109. https://doi.org/10.1186/s40478-022-01411-xCrossRefPubMedPubMedCentral Reinecke D, von Spreckelsen N, Mawrin C, Ion-Margineanu A, Fürtjes G, Jünger ST, Khalid F, Freudiger CW, Timmer M, Ruge MI, Goldbrunner R, Neuschmelting V (2022) Novel rapid intraoperative qualitative tumor detection by a residual convolutional neural network using label-free stimulated Raman scattering microscopy. Acta Neuropathol Commun 10:109. https://​doi.​org/​10.​1186/​s40478-022-01411-xCrossRefPubMedPubMedCentral
30.
go back to reference Schneider M, Vollmer L, Potthoff A-L, Ravi VM, Evert BO, Rahman MA, Sarowar S, Kueckelhaus J, Will P, Zurhorst D (2021) Meclofenamate causes loss of cellular tethering and decoupling of functional networks in glioblastoma. Neuro-oncology Schneider M, Vollmer L, Potthoff A-L, Ravi VM, Evert BO, Rahman MA, Sarowar S, Kueckelhaus J, Will P, Zurhorst D (2021) Meclofenamate causes loss of cellular tethering and decoupling of functional networks in glioblastoma. Neuro-oncology
32.
go back to reference Shapson-Coe A, Januszewski M, Berger DR, Pope A, Wu Y, Blakely T, Schalek RL, Li PH, Wang S, Maitin-Shepard J, Karlupia N, Dorkenwald S, Sjostedt E, Leavitt L, Lee D, Bailey L, Fitzmaurice A, Kar R, Field B et al (2021) A connectomic study of a petascale fragment of human cerebral cortex. bioRxiv, p 2021.05.29.446289. https://doi.org/10.1101/2021.05.29.446289CrossRef Shapson-Coe A, Januszewski M, Berger DR, Pope A, Wu Y, Blakely T, Schalek RL, Li PH, Wang S, Maitin-Shepard J, Karlupia N, Dorkenwald S, Sjostedt E, Leavitt L, Lee D, Bailey L, Fitzmaurice A, Kar R, Field B et al (2021) A connectomic study of a petascale fragment of human cerebral cortex. bioRxiv, p 2021.05.29.446289. https://​doi.​org/​10.​1101/​2021.​05.​29.​446289CrossRef
33.
go back to reference Ståhl PL, Fredrik S, Sanja V, Anna L, Fernández NJ, Jens M, Stefania G, Michaela A, Westholm JO, Mikael H, Annelie M, Sten L, Simone C, Åke B, Fredrik P, Igor CP, Pelin S, Jan M, Olaf B et al (2016) Visualization and analysis of gene expression in tissue sections by spatial transcriptomics. Science 353:78–82. https://doi.org/10.1126/science.aaf2403CrossRefPubMed Ståhl PL, Fredrik S, Sanja V, Anna L, Fernández NJ, Jens M, Stefania G, Michaela A, Westholm JO, Mikael H, Annelie M, Sten L, Simone C, Åke B, Fredrik P, Igor CP, Pelin S, Jan M, Olaf B et al (2016) Visualization and analysis of gene expression in tissue sections by spatial transcriptomics. Science 353:78–82. https://​doi.​org/​10.​1126/​science.​aaf2403CrossRefPubMed
34.
go back to reference Steiner G, Galli R, Preusse G, Michen S, Meinhardt M, Temme A, Sobottka SB, Juratli TA, Koch E, Schackert G, Kirsch M, Uckermann O (2022) A new approach for clinical translation of infrared spectroscopy: exploitation of the signature of glioblastoma for general brain tumor recognition. J Neurooncol. https://doi.org/10.1007/s11060-022-04204-3 Steiner G, Galli R, Preusse G, Michen S, Meinhardt M, Temme A, Sobottka SB, Juratli TA, Koch E, Schackert G, Kirsch M, Uckermann O (2022) A new approach for clinical translation of infrared spectroscopy: exploitation of the signature of glioblastoma for general brain tumor recognition. J Neurooncol. https://​doi.​org/​10.​1007/​s11060-022-04204-3
35.
go back to reference Straehle J, Erny D, Neidert N, Heiland DH, El Rahal A, Sacalean V, Steybe D, Schmelzeisen R, Vlachos A, Mizaikoff B, Reinacher PC, Coenen VA, Prinz M, Beck J, Schnell O (2022) Neuropathological interpretation of stimulated Raman histology images of brain and spine tumors: part B. Neurosurg Rev 45:1721–1729. https://doi.org/10.1007/s10143-021-01711-1CrossRefPubMed Straehle J, Erny D, Neidert N, Heiland DH, El Rahal A, Sacalean V, Steybe D, Schmelzeisen R, Vlachos A, Mizaikoff B, Reinacher PC, Coenen VA, Prinz M, Beck J, Schnell O (2022) Neuropathological interpretation of stimulated Raman histology images of brain and spine tumors: part B. Neurosurg Rev 45:1721–1729. https://​doi.​org/​10.​1007/​s10143-021-01711-1CrossRefPubMed
36.
go back to reference Stupp R, Taillibert S, Kanner A, Read W, Steinberg DM, Lhermitte B, Toms S, Idbaih A, Ahluwalia MS, Fink K, Di Meco F, Lieberman F, Zhu J-J, Stragliotto G, Tran DD, Brem S, Hottinger AF, Kirson ED, Lavy-Shahaf G et al (2017) Effect of tumor-treating fields plus maintenance temozolomide vs maintenance temozolomide alone on survival in patients with glioblastoma: a randomized clinical trial. JAMA 318:2306–2316. https://doi.org/10.1001/jama.2017.18718CrossRefPubMedPubMedCentral Stupp R, Taillibert S, Kanner A, Read W, Steinberg DM, Lhermitte B, Toms S, Idbaih A, Ahluwalia MS, Fink K, Di Meco F, Lieberman F, Zhu J-J, Stragliotto G, Tran DD, Brem S, Hottinger AF, Kirson ED, Lavy-Shahaf G et al (2017) Effect of tumor-treating fields plus maintenance temozolomide vs maintenance temozolomide alone on survival in patients with glioblastoma: a randomized clinical trial. JAMA 318:2306–2316. https://​doi.​org/​10.​1001/​jama.​2017.​18718CrossRefPubMedPubMedCentral
37.
go back to reference Ting JT, Kalmbach B, Chong P, Frates R, Keene CD, Gwinn RP, Cobbs C, Ko AL, Ojemann JG, Ellenbogen RG (2018a) A robust ex vivo experimental platform for molecular-genetic dissection of adult human neocortical cell types and circuits. Sci Rep 8:8407CrossRefPubMedPubMedCentral Ting JT, Kalmbach B, Chong P, Frates R, Keene CD, Gwinn RP, Cobbs C, Ko AL, Ojemann JG, Ellenbogen RG (2018a) A robust ex vivo experimental platform for molecular-genetic dissection of adult human neocortical cell types and circuits. Sci Rep 8:8407CrossRefPubMedPubMedCentral
41.
go back to reference Witcher MR, Park YD, Lee MR, Sharma S, Harris KM, Kirov SA (2010) Three-dimensional relationships between perisynaptic astroglia and human hippocampal synapses. Glia 58:572–587PubMedPubMedCentral Witcher MR, Park YD, Lee MR, Sharma S, Harris KM, Kirov SA (2010) Three-dimensional relationships between perisynaptic astroglia and human hippocampal synapses. Glia 58:572–587PubMedPubMedCentral
42.
43.
go back to reference Yamasaki T, Maekawa T, Fujita T, Tobimatsu S (2017) Connectopathy in autism spectrum disorders: a review of evidence from visual evoked potentials and diffusion magnetic resonance imaging. Front Neurosci 11:627CrossRefPubMedPubMedCentral Yamasaki T, Maekawa T, Fujita T, Tobimatsu S (2017) Connectopathy in autism spectrum disorders: a review of evidence from visual evoked potentials and diffusion magnetic resonance imaging. Front Neurosci 11:627CrossRefPubMedPubMedCentral
Metadata
Title
Technical report: surgical preparation of human brain tissue for clinical and basic research
Authors
J. Straehle
V. M. Ravi
D.H. Heiland
C. Galanis
M. Lenz
Junyi Zhang
N.N Neidert
A. El Rahal
I. Vasilikos
P. Kellmeyer
C. Scheiwe
J.H. Klingler
C. Fung
A. Vlachos
J. Beck
O. Schnell
Publication date
06-05-2023
Publisher
Springer Vienna
Keyword
Epilepsy
Published in
Acta Neurochirurgica / Issue 6/2023
Print ISSN: 0001-6268
Electronic ISSN: 0942-0940
DOI
https://doi.org/10.1007/s00701-023-05611-9

Other articles of this Issue 6/2023

Acta Neurochirurgica 6/2023 Go to the issue