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J Vis Exp. 2019 Jul 19;(149). doi: 10.3791/59511.

Comprehensive Autopsy Program for Individuals with Multiple Sclerosis.

Author information

1
Department of Neurosciences, Lerner Research Institute, Cleveland Clinic.
2
Department of Neurosciences, Lerner Research Institute, Cleveland Clinic; Mellen Center for Treatment and Research in Multiple Sclerosis, Neurological Institute, Cleveland Clinic.
3
Mellen Center for Treatment and Research in Multiple Sclerosis, Neurological Institute, Cleveland Clinic.
4
Department of Biomedical Engineering, Lerner Research Institute, Cleveland Clinic.
5
Department of Neurosciences, Lerner Research Institute, Cleveland Clinic; trappb@ccf.org.
#
Contributed equally

Abstract

We describe a rapid tissue donation program for individuals with multiple sclerosis (MS) that requires scientists and technicians to be on-call 24/7, 365 days a year. Participants consent to donate their brain and spinal cord. Most patients were followed by neurologists at the Cleveland Clinic Mellen Center for MS Treatment and Research. Their clinical courses and neurological disabilities are well-characterized. Soon after death, the body is transported to the MS Imaging Center, where the brain is scanned in situ by 3 T magnetic resonance imaging (MRI). The body is then transferred to the autopsy room, where the brain and spinal cord are removed. The brain is divided into two hemispheres. One hemisphere is immediately placed in a slicing box and alternate 1 cm-thick slices are either fixed in 4% paraformaldehyde for two days or rapidly frozen in dry ice and 2-methylbutane. The short-fixed brain slices are stored in a cryopreservation solution and used for histological analyses and immunocytochemical detection of sensitive antigens. Frozen slices are stored at -80 °C and used for molecular, immunocytochemical, and in situ hybridization/RNA scope studies. The other hemisphere is placed in 4% paraformaldehyde for several months, placed in the slicing box, re-scanned in the 3 T magnetic resonance (MR) scanner and sliced into centimeter-thick slices. Postmortem in situ MR images (MRIs) are co-registered with 1 cm-thick brain slices to facilitate MRI-pathology correlations. All brain slices are photographed and brain white-matter lesions are identified. The spinal cord is cut into 2 cm segments. Alternate segments are fixed in 4% paraformaldehyde or rapidly frozen. The rapid procurement of postmortem MS tissues allows pathological and molecular analyses of MS brains and spinal cords and pathological correlations of brain MRI abnormalities. The quality of these rapidly-processed postmortem tissues (usually within 6 h of death) is of great value to MS research and has resulted in many high-impact discoveries.

PMID:
31380830
DOI:
10.3791/59511

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