[Romanowsky dyes and the Romanowsky-Giemsa effect. 3. Microspectrophotometric studies of Romanowsky-Giemsa staining. Spectroscopic evidence of a DNA-azure B-eosin Y complex producing the Romanowsky-Giemsa effect].

摘要: The Romanowsky-Giemsa staining (RG staining) has been studied by means of microspectrophotometry using various conditions. As cell material we employed in our model experiments mouse fibroblasts, LM cells. They show a distinct pattern. RG was performed with the chemical pure dye stuffs azure B and eosin Y. In addition stained cells separately or Staining parameters were pH value, concentration, time etc. Besides normal also after RNA DNA digestion. spectra species measured self constructed microspectrophotometer photon counting technique. optical ray pass diagramm electronics are briefly discussed. nucleus cells, congruent to 7, is purple, cytoplasm blue. After digestion purple respectively blue coloration completely disappeares. Therefore preferentially biological substrates. spectrum nuclei, three absorption bands distinguishable: A1 (15400 cm-1, 649 nm), A2 (16800 595 nm) DNA-bound monomers dimers RB (18100 552 intense Romanowsky band. Our extensive experimental shows clearly that produced complex DNA, higher polymers (degree association p greater than 2) primarily held together electrostatic interaction: inding polymer cations polyanion generates positively charged binding sites DNA-azure which subsequently occupied Y anions. It can be spectroscopically shown electronic states attached interact. By this interaction red shifted shifted. both molecular overlap generate Its strong maximum at 18100 cm-1 due part B-eosin complex. discussed shift main reason for nuclei. Using special technique it possible prepare an artificial calf thymus as nucleic acid two Y.(ABSTRACT TRUNCATED AT 400 WORDS)