Stable Sudanophilia of Human Neutrophil Leucocytes in Relation to Peroxidase and Oxidase

Abstract

It seems improbable that leucocyte oxidase is a fatty acid peroxide, as suggested by Sehrt, on these grounds: 1. Positive reactions for aldehyde and for ethylene groups are not obtained. Hence the presence of fatty acid peroxides in leucocytes seems unlikely. 2. Peroxidase has been isolated from leucocytes by Agner. Histochemical behavior on treatment with adverse reagents and with heat indicates similarity of benzidine peroxidase and of indophenol oxidase, but dissimilarity of these from the sudanophil substance in leucocytes. 3. Sudan dyes demonstrate or produce in neutrophil leucocytes deeply colored granules of quite variable size, and the size of the granules apparently increases with the duration of staining. 4. The demonstration of sudanophil granules in leucocytes is accomplished with difficulty. The colored granules are highly stable toward many Sudan dye solvents. Decolorization is difficult and recoloration is uncertain. 5. True fats and lipoids, after staining with Sudan dyes, are readily decolorized by appropriate dye solvents and are restainable in apparently undiminished amounts. 6. Some oxidants tend to impair sudanophilia of leucocytes. Potassium bichromate improves the preservation of sudanophil lipoids, as in the Ciaccio method, but destroys the sudanophilia of polymorphonuclear leucocytes. 7. Hydrogen peroxide apparently possesses an inconstant and irregular, though sometimes quite pronounced, accelerating effect on the production of Sudan stained granules in leucocytes. 8. The sudanophilia of leucocytes depends on an as yet unexplained chemical combination of the dyes with cytoplasmic constituents which are probably not in the form of preexisting granules, rather than on lipoid staining.

What to do with high autofluorescence background in pancreatic tissues – an efficient Sudan black B quenching method for specific immunofluorescence labelling

“…Clearly, Sudan black B quenched autofluorescence of acinar cells effectively, which was probably mediated by the ability to stain zymogen granules intensively (Figure ), thereby obscuring the autofluorescent components. Its property to stain intracellular granules has long been known for leucocytes, dermal mast cells and also for pancreatic B cells . In line with these observations, more recent studies confirmed that Sudan black B is also able to mask autofluorescence of myeloid granules .…”

“…Its property to stain intracellular granules has long been known for leucocytes, dermal mast cells and also for pancreatic B cells. [34][35][36][37] In line with these observations, more recent studies confirmed that Sudan black B is also able to mask autofluorescence of myeloid granules. 13 In pancreatic tissue we found that Sudan black B further stains connective tissue fibres and erythrocytes intensively ( Figure 3H), which clearly supported the quenching of tissue autofluorescence and the intense fluorescence caused by the presence of haemoglobin.…”

“…Sudan black B stains a wide range of subcellular molecular structures, above all lipids and phospholipids, but also some proteins and mucopolysaccharides. The chemical interactions between the dye and tissue components can be manifold, and discussion in the literature regarding these interactions is controversial . In pancreatic tissue, we identified nuclei and zymogen granules of acinar cells to exhibit notably bright autofluorescence, besides more ubiquitous tissue components such as connective fibres and erythrocytes (Figures and ).…”

“…The chemical interactions between the dye and tissue components can be manifold, and discussion in the literature regarding these interactions is controversial. 24,25,33,34 In pancreatic tissue, we identified nuclei and zymogen granules of acinar cells to exhibit notably bright autofluorescence, besides more ubiquitous tissue components such as connective fibres and erythrocytes (Figures 1 and 3). Clearly, Sudan black B quenched autofluorescence of acinar cells effectively, which was probably mediated by the ability to stain zymogen granules intensively (Figure 2), thereby obscuring the autofluorescent components.…”

Partial and Severe Peroxidase and Phospholipid Deficiency in Eosinophils

“…positive. The 4 cases of hydrocephaly tested by us showed [15] state that there is no connection between peroxidase and phospholipids; however, in all cases we have previously described [20,23] as well as the present, the parallelism between the reaction of the 2 substances is very evident. Similar parallelism is found in polynuclear neutrophils in the Alius-Grignaschi anomaly [25].…”