Isolation and biochemical mapping in the range of 10 −7 to 10 −12 g of fresh, single mammalian neurons in brain I. Techniques

• Published in: TrAC, Trends in analytical chemistry (Regular ed.) Vol. 14; no. 4; pp. 141 - 148
• Main Author: HYDEN, H
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier B.V 01-04-1995; Elsevier Science
• Subjects: Biological and medical sciences; Fundamental and applied biological sciences. Psychology; Isolated neuron and nerve. Neuroglia; Vertebrates: nervous system and sense organs; RNA; Enzyme; Neuroglia; Central nervous system; Biological macromolecule; Review; K; Microassay; Proteins; Vertebrata; Mammalia; Na; Neuron; Enzymatic activity; Preparation; Plasma membrane; Analytical method; Hydrolases; Isolation; exchanging ATPase; Brain (vertebrata)
• ISSN: 0165-9936; 1879-3142
• DOI: 10.1016/0165-9936(95)98310-5

A free-hand technique to remove single, fresh nerve cells including 50–100 μm of their dendrites, and the removal of its surrounding glia from a defined locus in the mammalian brain is described. Exposition of the neuronal post-synaptic areas, removal of the neuronal nucleus and preparation of a functional plasma membrane are likewise described. Biophysical and biochemical micro methods for determination of macro molecules from 20 pg and of ion fluxes are described. These include UV micro-photometry, micro-electrophoresis (in 20 μm cellulose gel fibers and in 215 or 440 μm gel-filled capillaries), analysis of radiolabeled proteins, and determination of ATPase activity, all applied to single cells or cell fragments.