Abstract
INTRODUCTIONThe eukaryotic cell has evolved to compartmentalize its functions and transport various metabolites among cellular compartments. Therefore, in cell biology, the study of organization and structure/function relationships is of great importance. Lysosomes are membranous sacs--diverse in shape and size--containing more than 40 different acid hydrolases. The enzymes operate optimally at the acidic pH (~5) within the lysosome to break down various substances. It is thought that the highly glycosylated nature of the proteins of the Golgi membrane helps to protect them from degradation. A number of the fluorescent approaches to visualizing lysosomes make use of their acidic pH. Commonly used probes include neutral red, N-(3-[2,4-dinitrophenyl amino] propyl)-N-(3-aminopropyl)methylamine (DAMP), and acridine orange (a DNA stain). This protocol describes the labeling of lysosomes in live cells with neutral red.
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