Fluorescence microscopy of viable mast cells stained with different concentrations of acridine orange

Abstract

Freshly harvested rat peritoneal mast cells were stained with different concentrations of acridine orange, a metachromatic fluorochrome known to form complexes with chromatin and muscopolysaccharides. Fluorescence metachromasia was observed in cytoplasmic granules in cell populations with intracelluar dye contents as low as 5 X 10(-16) mole per cell, one-half decade lower than required to produce metachromatic staining of the nucleus. Cytoplasmic granules did not stain uniformly throughout the cell; some granules exhibited red fluorescence and others green. As the amount of acridine orange uptake per cell was increased, cytoplasmic fluorescence became uniformly red and nuclear fluorescence gradually changed from green to yellow.

Structural studies on the membrane-bound immunoglobulin E-receptor complex. 1. Characterization of large plasma membrane vesicles from rat basophilic leukemia cells and insertion of amphipathic fluorescent probes

“…of the membrane appears to be in vesicles greater than 0.1 nm in diameter. Although these vesicles appear to enclose trapped cytoplasm ; also, see below), included organelles are generally not observed when fluorescent stains such as rhodamine 123 for mitochondria (Johnson et al, 1980) and acridine orange for histamine-containing granules (Love, 1979) are used.…”

Quantitative measurement of mast cell degranulation using a novel flow cytometric annexin-V binding assay

“…The measurement of mast cell degranulation has been largely limited to the monitoring enzyme activity or the levels of a particular mediator released by a population of cells. Although methods have been used to monitor mast cell degranulation using fluorescent probes such as acri-dine orange (15), TMA-DPH (16), and berberine (17), they have suffered from a lack of specificity for the exocytic process or poor signal-to-noise ratios.…”

Updating Ortho- and Metachromatic Acridine Orange Fluorescence in Cytochemical Chromosome Staining: A Proposal for Understanding Its Differential Fluorescence on Double- and Single-Stranded Nucleic Acids Substrates Based on Intercalation

“…In particular, the interaction of AO with biological substrates in the living or dead cell leads to such ortho-/metachromatic phenomenon as a consequence of the local concentration of the fluorophore and, therefore, its aggregation state. A schematic microscopic correlation of AO change in fluorescence wavelength for different cell subdomains and increasing concentrations of the fluorophore is clearly observed in Figure 4, where the nuclei and heparin-containing granules of mast cells are used as an illustrative staining model [17]. At low AO concentrations, all emission is orthochromatic (green), so there is no differential labeling.…”

“…In addition to NAs, numerous polyanionic substrates in fixed tissue or cell samples exhibit metachromatic fluorescence when stained with AO. Examples are chondroitin sulfates from the cartilage matrix, the mucin content of intestinal goblet cells, and specific heparin-containing granules of mast cells [17]. Likewise, on account of their acridine core fluorophore, other colorants such as acriflavine, acridine yellow G, coriphosphine O, euchrysin GGNX, phosphine E, etc., also display similar metachromatic properties [5,7,18].…”