Separation of Neuronal and Glial Cells and Subcellular Constituents

Publisher Summary This chapter focuses on the separation of neuronal and glial cells and subcellular constituents. The analysis of any tissue usually proceeds at two distinct levels. One is an anatomical investigation in which the tissue structure is examined with ever-increasing resolution. The other is a biochemical examination in which the molecular components of the tissue are examined and their organization into units of structure is subsequently investigated. Cellular neurobiology aims to understand the diverse cells of the central nervous system (CNS) and how they work together to process, store, and utilize information. To obtain an idea of the potentialities of various brain cells, it would be desirable to isolate relatively pure samples of specific groups of neuronal and glial cells and study their function. The principal steps in cell isolation involve methods for disaggregating cells and separating them into relatively homogeneous fractions. The issues involved in tissue disaggregation include the media used in preparation of the tissue, incubation or lack of it, and the mechanical or enzymatic means for dissociating cells. The problem of contamination is present not only in cellular CNS fractions but also in subcellular fractions derived from brain. All the general cell organelles, namely, nuclei, mitochondria, microsomes, ribosomes, lysosomes, and plasma membrane preparations, if derived from whole brain, are bound to be a heterogeneous collection from very different types of cells. The chapter discusses three methods that appear to offer promise of finding more specific cell isolation procedures: (1) affinity chromatography, (2) the use of an automated cytofluorograph to sort cells, and (3) the use of magnetic microspheres to separate cells. All these three methods utilize the specificity of defined antibodies to identify the cell or particle in question. The isolation of the particle takes place directly by means of the binding in affinity systems.

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