On the nature of Romanowsky-Giemasa staining and its significance for cytochemistry and histochemistry: an overall view

Abstract

The chances of Romanowsky---Giemsa (RG) staining becoming a reliable and useful histochemical procedure are reviewed, based on the now proven fact that RG staining requires two dyes only, namely, cationic Azure B and anionic Eosin Y. These two dyes differ from otherwise similar dye combinations in that they give, on distinct biological substrates, one additional colour, purple, which cannot be obtained by the use of either dye alone. The purple colour characterizes the Romanowsky--Giemsa effect (RGE), which is the essential feature of RG staining. Consideration is given to the physico-chemical and morphological implications of RGE. Of primary importance is the nature of the biological substrates where RGE occurs, and also of those where it has never been observed. The way substrates react to RG stains largely depends on the kind of pretreatment they have received; for instance, alcoholic fixation preserves RGE but formaldehyde may inhibit it. Physico-chemical factors are considered which, by altering either the biological substrates or the composition of the staining solutions, may modify the RG staining pattern. This review also serves as an introduction for a series of experimental papers that will follow and which are intended to consolidate the basis of RG staining, a method which holds much promise as a useful histochemical tool.

Giemsa as a fluorescent stain for mineralized bone.

“…The effect of differential staining of blood cells (the so-called Romanowsky effect) is the best known chromatic effect afforded by Giemsa and Giemsa-related solutions (6). Eosin Y, one component of Giemsa and Giemsa-like solutions, acts as both a diachrome and a fluorochrome ( 5 ) .…”

The Heterochromatin Condensation State in Peripheral “Gene Poor” and Central “Gene Rich” Nuclear Regions of Less Differentiated and Mature Human Leukemic Cells: A Mini-Review with Additional Original Observations

“…It is possible to measure the heterochromatin density in digital images of stained cells by the polychrome procedure after the conversion of captured color images to gray scale using the red channel (NIH Image Program, Scion for Windows, Scion Corp., USA). The polychrome May-Grünwald -Giemsa-Romanowsky staining procedure is useful not only for the cell identification including differentiation and maturation stages but also for .the heterochromatin density measurements [1,[4][5][6]20]. The calculated arbitrary density units facilitate to estimate the ratio of the mean heterochromatin condensation state in central to peripheral nuclear regions (Figure 1).…”

Recommandations techniques et règles de bonne pratique pour la coloration de May-Grünwald-Giemsa : revue de la littérature et apport de l’assurance qualité

“…Les principes de la coloration de MGG, issus des travaux de Chenzinsky et de Romanowsky sur les protozoaires (1891), ont été repris par Wright, May, Grünwald, Giemsa et Pappenheim (1902-14), puis développés par Lopez Cardozo (1973) et par Wittekind (1983) [4][5][6][7][8][9]24]. Comme le rappelle Lopez Cardozo [18], la découverte de la coloration provient d'une erreur de laboratoire en 1890 : le bleu de méthylène de la coloration de Löffler (diagnostic de malaria) ayant vieilli et s'étant oxydé, Chenzinsky et Romanowsky notèrent que les noyaux du Plasmodium malariae étaient devenus rouge pourpre sur les frottis sanguins [4,18,25].…”

“…. Comme le suggérait Lopez Cardozo [18], une coloration simple (un seul colorant) n'est pas obligatoirement plus mauvaise qu'une polychrome, et il faut se souvenir que tous les dérivés azurés du bleu de méthylène sont polychromes par oxydation en solution [24][25][26][27][28][38][39][40].…”