Abstract
A simple and reproducible method for cryo-preservation of brain tissue from patients with Alzheimer's disease is described. Fresh brain slices (1 cm thick) obtained less than 6 h postmortem are placed in sealed plastic bags, sandwiched between 0.3-cm-thick aluminium sheets, and frozen by placing the entire “sandwich” between layers of dry ice pellets. The frozen brain slices are stored at −85 °C. Specific anatomic areas can be retrieved at any time for light and electron microscopic, immunocytochemical, autoradiographic and neurochemical studies.
Article PDF
Avoid common mistakes on your manuscript.
References
Earle KE (circa (1964) Examination of the brain (necropsy technique). American Registry of Pathology. Armed Forces Institute of Pathology, Washington, D.C.
Glauert AM (1984) Fixation, dehydration and embedding of biological specimens. In: Glauert AM (ed) Practical methods in electron microscopy, 1st edn vol 3, part I. Elsevier North-Holland, New York
Heuser JE, Reese TS, Landis DMD (1976) Preservation of synaptic structure by rapid freezing. Cold Spring Harbor Symp Quant Biol 40:17–24
Hsu S-M, Raine L, Fanger H (1981) Use of avidin biotin-peroxidase complex (ABC) in immunoperoxidase techniques: a comparison between ABC and unlabeled antibody (PAP) procedures. J Histochem Cytochem 29:577–580
Kitamoto T, Ogomori K, Tateishi J and Pruasiner SB (1987) Formic acid pretreatment enhances immunostaining of cerebral and systemic amyloids. Lab Invest 57:230–236
Luna LG (1968) Manual of histologic staining methods of the Armed Forces Institute of Pathology. American Registry of Pathology, 3rd edn, McGraw-Hill, New York
Masters CL, Gajdusek DC, Gibbs CJ (1981) Creutzfeldt-Jakod disease virus isolations from the Gerstmann-Sträussler syndrome. With an analysis of the various forms of amyloid deposition in the virus-induced spongiform encephalopathies. Brain 104:559–588
Smith TW, Anwer U, DeGirolami U, Drachman DA (1987) Vacuolar change in Alzheimer's disease. Arch Neurol 44:1225–1228
Sternberger LW (1986) Immunocytochemistry, 3rd edn. Wiley, New York
Terracio L, Bankston PW, McAteer JA (1981) Ultrastructural observations on tissues processes by a quick-freezing, rapiddrying method: comparison with conventional specimen preparation. Cryobiology 18:55–71
Tian M, Reger JF, Armstrong WE (1991) Electron microscopic and immunocytochemical study of rapidly frozen, freezesubstituted neural lobes of rats. J Neurocytol 20:79–96
Trump BF, Young DE, Arnold EA, Stowell RE (1965) Effects of freezing and thawing on the structure, chemical constitution, and function of cytoplasmic structures. Fed Proc 24:S144-S168
van Harreveld A (1966) Brain tissue electrolytes. Investigation of water and electrolyte distribution by electron microscopy. In: Bittar EE (ed) Molecular biology and medicine series. Butterworths, Washington pp 127–158
van Harreveld A, Crowell J (1964) Electron microscopy after rapid freezing on a metal surface and substitution fixation. Anat Rec 149:381–386
van Harreveld A, Crowell J, Malhotra SK (1965) A study of extracellular space in central nervous tissue by freeze-substitution. J Cell Biol 25:117–137
Yamaguchi H, Hirai S, Morimatsu M, Shoji M, Harigaya Y (1988) Diffuse type of senile plaques in the brains of Alzheimer-type dementia. Acta neuropathol 77:113–119
Zierold K (1986) Preparation of cryosections for biological microanalysis. In: Muller M, Berker RP, Boyde A, Wolosewick JJ (eds) The science of biological specimen preparation. SEM Inc AMF O'Hare, Chicago, pp 119–127
Author information
Authors and Affiliations
Additional information
Supported by grants NIA# AG05136 and AG06781-06 to the University of Washington Alzheimer's Disease Research Center and Alzheimer's Disease Patient Registry
Rights and permissions
About this article
Cite this article
Nochlin, D., Mackenzie, A.P., Bryant, E.M. et al. A simple method of rapid freezing adequately preserves brain tissue for immunocytochemistry, light and electron microscopic examination. Acta Neuropathol 86, 645–650 (1993). https://doi.org/10.1007/BF00294305
Received:
Revised:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00294305