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Publisher Summary This chapter discusses the methods for the separation of pancreatic islet cells according to functional characteristics. The physical properties of cells, such as their size and density, have served as a basis for their isolation. The ability to isolate cells according to the functional characteristics would permit a more precise cellular analysis of the tissue function, and it could also provide the appropriate in vitro models for investigating the physiopathological basis of certain diseases. In the study of diabetes mellitus, it is not clear whether and how the various pancreatic islet cells cooperate in the generation of the in vivo insulin response. A separation of the various islet cell types and their further distribution into the functionally homogeneous subpopulations may provide an experimental approach to the questions. The autofluorescence-activated cell sorting permits the isolation of islet cells with similar size and density, but with a different metabolic responsiveness to their physiological regulator glucose. The chapter reviews the work on islet cells and addresses investigations involved in diabetes research. It also intends for cell biologists in search of ways to isolate cells according to functional criteria.

Publisher Summary This chapter reviews the technique of autofluorescence-activated cell sorting and discusses its usefulness in recognizing and isolating cell preparations with different functional properties. Earlier studies demonstrated how the measurement of the cellular FAD and NAD(P)H fluorescent intensities allows the distinction and isolation of various types of pancreatic endocrine cells. Autofluorescence-activated cell sorting permitted the preparation of pure and single insulin-containing B cells, which were used to investigate the functional properties of this cell type without interference from other cell types. In the course of these studies, it was noticed that this endocrine cell population does not represent a homogeneous group of identical cells, but rather is composed of functionally diverse subpopulations. Several of these subpopulations have been recognized during flow cytometric analysis, whereby the cellular autofluorescent compounds FAD and NAD(P)H appeared again to be particularly powerful discriminators. The observation that a pure pancreatic B-cell population is composed of functionally diverse subpopulations underlines the need to purify cells according to their functional properties rather than according to their structural characteristics.