An improved approach to prepare human brains for research.

Author(s): Vonsattel JP, Aizawa H, Ge P, DiFiglia M, McKee AC, MacDonald M, Gusella JF, Landwehrmeyer GB, Bird ED, Richardson EP Jr

Publication: J Neuropathol Exp Neurol, 1995, Vol. 54, Page 42-56

PubMed ID: 7815079 PubMed Review Paper? No

Purpose of Paper

Conclusion of Paper

Studies

1. Study Purpose

   More than 3,000 brains were collected after autopsy and differentially preserved over a 15 year period as a collaboration between the Brain Tissue Resource Center and the Massachusetts Alzheimer Disease Research Center. Specimens were frozen on dry ice directly or using a cooling device, or snap-frozen by immersion in liquid nitrogen, liquid nitrogen vapor, or liquid nitrogen-cooled isopentane and analyzed for morphological integrity. Postmortem intervals ranged between 0.3 to 145 hours.

   Summary of Findings:

   Brain specimens frozen by immersion in liquid nitrogen vapor resulted in excellent preservation of gross and microscopic morphology with no apparent freeze-induced artifacts. Comparatively, coronal brain slices frozen on dry ice with a cooling device for 3-5 minutes adequately preserved morphology with minor artifacts, such as intracellular and extracellular holes, while specimens frozen by immersion in liquid nitrogen or liquid nitrogen-cooled isopentane displayed macroscopic cracks. Halved brain specimens frozen intact on dry ice displayed a significant number of artifacts that impaired further dissection and prevented cell type identification.

   Biospecimens

   • Tissue - Brain

   Preservative Types

   • Frozen

   Diagnoses:

   • Normal
   • Parkinson's Disease
   • Alzheimer's Disease
   • Lewy Body Disease
   • Other diagnoses
   • Autopsy

   Platform:

   Analyte	Technology Platform
   Morphology	Macroscopic observation
   Morphology	H-and-E microscopy

   Pre-analytical Factors:

   Classification	Pre-analytical Factor	Value(s)
   Biospecimen Preservation	Type of fixation/preservation	Frozen Snap frozen
   Biospecimen Preservation	Cooling or freezing method/ rate	Dry ice Cooling device Pre-cooled isopentane Liquid nitrogen Liquid nitrogen vapor

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2. Study Purpose

   More than 3,000 brains were collected after autopsy and differentially preserved over a 15 year period as a collaboration between the Brain Tissue Resource Center and the Massachusetts Alzheimer Disease Research Center. Specimens were frozen on dry ice directly or using a cooling device, or snap-frozen in liquid nitrogen and analyzed for mRNA and protein expression. Postmortem intervals ranged between 0.3 and 145 hours.

   Summary of Findings:

   Coronal brain specimens frozen using a cooling device constructed of aluminum plates placed on dry ice for 3-5 minutes were of suitable quality for immunohistochemistry and in situ hybridization analyses. In situ hybridization and immunohistochemistry was also successful with specimens frozen in liquid nitrogen vapor. Halved brain specimens frozen intact with dry ice were suitable for DNA, RNA, and protein expression analyses, although compromised cellular morphology prevented expression localization.

   Biospecimens

   • Tissue - Brain

   Preservative Types

   • Frozen

   Diagnoses:

   • Not specified

   Platform:

   Analyte	Technology Platform
   RNA	In situ hybridization
   Protein	Immunohistochemistry

   Pre-analytical Factors:

   Classification	Pre-analytical Factor	Value(s)
   Biospecimen Preservation	Type of fixation/preservation	Frozen Snap frozen
   Preaquisition	Postmortem interval	0.3-145 h
   Biospecimen Preservation	Cooling or freezing method/ rate	Cooling device Dry ice

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