Selective fluorescence reaction of indigocarmine stained eosinophil leucocyte granules induced by alkaline reduction of the bound dye to its leuco derivative

Production of superoxide radical in reductive metabolism of a synthetic food-coloring agent, indigocarmine, and related compounds

“…The reducibility of indigocarmine by the enzyme is extremely high compared with other biological reductions (McLane et al, 1983). There is a report that a reduction product of indigocarmine was formed on treatment with sodium hydrosulfite or other reducing agents, and the absorbance peak of indigocarmine disappeared (Stockert and Trigoso, 1994). We observed the same spectral change upon addition of sodium hydrosulfite and upon metabolic reaction with rat liver microsomes.…”

“…In contrast, Kitamura and Tatsumi (1990) reported that various a,h-ketoalkenes were reduced to the corresponding ketoalkanes by rat liver cytosolic double-bond reductase. Indigocarmine is known to be chemically reduced to the double-bond-reduced compound, the leuco form, by mild reductants such as sodium hydrosulfite (Stockert and Trigoso, 1994). However, there is no report of enzymatic reduction of indigocarmine.…”

Fluorescence of the natural dye saffron: Selective reaction with eosinophil leucocyte granules

“…After liberation of carboxyl groups, the hydrolyzed derivative of saffron (crocetin) could function as an anionic dye (Lillie 1977) with affinity for acidophilic proteins such as those of eosinophil granules (Stockert et al 1991;Stockert and Trigoso 1994). Although far from complete, the observations described here suggest the possibility of employing the old microscopic stain saffron as a new and useful fluorochrome.…”

“…Smears of chicken, rat, human and horse peripheral blood were fixed in methanol for 2 rain and air-dried, Horse blood was chosen on account of the large size of the eosinophil granules , which facilitates their detailed visualization and a more precise recognition of selective fluorescence reactions (Stockert et al 1991;1994). Chicken blood smears were also used because of the highly acidophilic nature of the beterophil granules (Schalm et al 1975;Hawkey and Dennett 1989).…”

Fluorescent cytochemistry of acid phosphatase and demonstration of fluid-phase endocytosis using an azo dye method

“…Therefore, there is a growing interest in converting classical staining procedures for bright field illumination into their corresponding counterparts for fluorescence microscopy (e.g., Stockert et al 1991;Espada et al 1993Stockert and Trigoso 1994). Likewise, it is highly desirable to develop new fluorescence methods for enzyme cytochemistry and immunocytochemistry (e.g., Dolbeare et al 1980;Raap 1986).…”