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The color purple: from royalty to laboratory, with apologies to Malachowski.

Department of Anatomy, Microbiology and Pathology, University of Minnesota, School of Medicine, Duluth, Minnesota, USA.
Biotechnic & Histochemistry (Impact Factor: 1). 02/2011; 86(1):7-35. DOI: 10.3109/10520295.2010.515490
Source: PubMed

ABSTRACT The components of the blood stain, eosin and methylene blue, were introduced by Baeyer and Caro, respectively. Methylene blue was used primarily for detecting Mycobacterium tuberculosis until Ehrlich in 1880 mixed methylene blue with acid fuchsin to produce what he termed a "neutral stain," which allowed differentiation of blood cells. Eight years later, Chęciń ski changed the acidic component of the dye to eosin. Plehn subsequently altered the proportions of eosin and methylene blue to produce a greater range of red and blue hues. In 1891, Malachowski and Romanowsky independently developed stains composed of eosin and "ripened" methylene blue that not only differentiated blood cells, but also demonstrated the nuclei of malarial parasites. A number of "ripening" or "polychroming" techniques were investigated by different groups, but the aqueous dye solutions produced were unstable and precipitated rapidly. Subsequently, methanol was introduced as a solvent for the dye precipitate and techniques were developed that utilized the fixative properties of the methanolic solution prior to aqueous dilution for staining. This avoided the troublesome process of heat fixation of blood films. Giemsa further improved these techniques by using more controlled methods of methylene blue demethylation. In addition, he used measured amounts of known dyes and increased dye stability by adding glycerol to the methanol solvent. With the outbreak of World War I, it became difficult to obtain German dyes outside of Germany; during the World War II, it became impossible. In their effort to improve the inferior American versions of Giemsa's stain, Lillie, Roe, and Wilcox discovered that the best staining results were obtained using pure methylene blue, one of its breakdown products (azure B) and eosin. These three substituents remain the major components of the stain to this day.

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    The 12th International Congress of Immunology and Allergy of Iran, Iran; 01/2014
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    ABSTRACT: Objectives: Malaria infection is a major public health problem and cause of morbidity and mortality particularly among children in tropical and subtropical regions of the world. The aim of this present study was to determine the effect of plasmodium parasitaemia on some haematological parameters of children visiting the children emergency unit of Sokoto State Specialist Hospital in Sokoto, North Western, Nigeria. Method: This study was conducted among 126 children aged 2-11 years with mean age 5.36 ± 2.50 years presenting to the children emergency unit of Sokoto Specialist Hospital with history of febrile illness. Out of the children studied, 66 (52.4%) were positive for malaria while 60 (47.6%) were negative. Haematological parameters were analyzed using Mythic 22 CT 5- part differential Haematology analyzer (Orphée, Switzerland). Testing for malaria was carried out using the Onset Malaria Plasmodium falciparum (Pf) antibody (Ab) rapid test (CTK Biotech, Inc. USA) and speciation and number of parasites per high field was carried out on the Giemsa stained thing blood film. Results: The mean PCV, haemoglobin and platelet count of plasmodium- parasitized children was significantly lower compared to un-infected controls (29.48, 10.36 and 188.68) versus (32.76, 11.34 and 327.50) respectively (p=0.01). The prevalence of anaemia and thrombocytopenia was significantly higher among Plasmodium parasitized subjects compared to non-parasitized controls. Plasmodium falciparum was the predominant specie among the parasitized subjects. A negative and significant correlation was observed between the high number of parasite per high field and platelet count as index of thrombocytopenia and haemoglobin as index of anaemia (r=0.62 and p=0.75) respectively (p= 0.01) among parasitized subjects. Plasmodium parasitaemia was more prevalent among children in the 2-5 years age group (52.4%) compared to children in the 6-11 years age group (47.6%). Male children were more predisposed to malaria (53.0%) compared to female children (47.0%). Conclusion: Plasmodium parasitaemia has a significant impact on the haemoglobin, packed cell volume and platelet count of malaria parasitized children in Sokoto, Nigeria. Preventative strategies including regular chemoprophylaxis, intermittent preventative treatment with antimalarials, provision of iron supplementation and insecticide-treated bed nets should be implemented urgently to prevent the negative impact of malaria parasitaemia on the haematological parameters of children in the area. There is need for community and peer-based awareness and education initiatives to strengthen the malaria prevention programme by educating parents on the benefits of effective environmental sanitation to destroy the breeding sites of Anopheles mosquito –the vector of malaria.
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    ABSTRACT: Mixtures of polychrome methylene blue-eosin Y (i.e., Giemsa stain) are widely used in biological staining. They induce a striking purple coloration of chromatin DNA (the Romanowsky-Giemsa effect), which contrasts with the blue-stained RNA-containing cytoplasm and nucleoli. After specific prestaining treatments that induce chromatin disorganization (giving banded or harlequin chromosomes), Giemsa staining produces a differential coloration, with C- and G-bands appearing in purple whereas remaining chromosome regions are blue. Unsubstituted (TT) and bromo-substituted (BT) DNAs also appear purple and blue, respectively. The same occurs in the case of BT and BB chromatids.In addition to discussing the use of Giemsa stain as a suitable method to reveal specific features of chromosome structure, some molecular processes and models are also described to explain Giemsa staining mechanisms of chromatin.
    Methods in molecular biology (Clifton, N.J.) 01/2014; 1094:25-38. DOI:10.1007/978-1-62703-706-8_3 · 1.29 Impact Factor

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