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At least 127 records · Page 7Linked to original sources
PubMed2005

Simultaneous differentiation between Theileria spp. and Babesia spp. on stained blood smear using PCR.

The tick-borne diseases of livestock constitute a complex of several diseases with different etiological agents. Theileriosis and babesiosis belong to this complex and are severe and often fatal protozoan tick-borne diseases of ruminants worldwide. This results in high economical losses yearly in Iran. The most common diagnostic method for the identification of piroplasms in Iran is Giemsa staining of blood smear, which is unspecific, accompanied by some technical problems and, in some cases, impossible, due to the carriers. In contrast, immunostaining is more specific and can only be performed with suitably prepared blood smears, but cannot be used also for the carriers. The most specific method for the differential diagnosis of piroplasms is the method of polymerase chain reaction. We extracted DNA from different sources of blood samples, including from already stained blood smears. The extracted DNA was subsequently amplified using specific primers derived from Theileria heat shock protein hsp70, Theileria lestoquardi ms1-2 gene, Babesia rhoptry protein gene and piroplasms hyper variable region V4 of 18S rRNA gene. The results show that it is possible to detect piroplasms in already stained blood smears as well enabling a simpler method to be developed for the collection of the samples. Furthermore, it is possible to analyse the already stained and registered blood smears from the patients with unclear differential diagnosis, e.g. in the carriers. In addition, the results revealed that using a primer designed from the hyper variable region V4 of 18S rRNA, it is possible to detect and differentiate simultaneously the genera Theileria and Babesia in DNA samples isolated from already stained blood smears.

Animals↗
PubMed2004

Atomic force microscopy of human metaphase chromosomes after differential staining of sister chromatids.

Human metaphase chromosomes, in which 5-bromo-deoxyuridine (BrdU) had been incorporated into the DNA, were treated with the fluorescent plus Giemsa (FPG) method. Use of this method distinctly stained one of the paired sister chromatids with the Giemsa solution due to the difference in content of BrdU in the two chromatids. These chromosomes with their differential staining of sister chromatids were observed by atomic force microscopy (AFM). In the air-dried specimens, one of the paired chromatids that was stained strongly with Giemsa solution was about two times higher than the counterpart that was stained faintly with Giemsa solution. In the critical point dried chromosomes, the height of the Giemsa positive chromatid roughly matched that of the Giemsa negative counterpart. These findings imply that the arrangement of the Giemsa negative chromatid after FPG staining is fragile and easily collapses due to the surface tension of water during air-drying. At higher magnifications, the surface structure differed between Giemsa positive and negative chromatids; the Giemsa positive chromatid (i.e., unifilarly BrdU-incorporated chromatid) was composed of fibrous structures while the Giemsa negative chromatid (i.e., bifilarly BrdU-incorporated chromatid) exhibited a fine granular appearance. These structural changes in the sister chromatids are thought to arise from the ultraviolet irradiation and heating of the chromosomes during FPG staining.

Antimetabolites↗
PubMed1999

Morphometry and quality control for a May-Grunwald Giemsa stained preparation. A 40 centers cooperative study.

A multicentric study including 40 medical centers has been done on May-Grunwald Giemsa (MGG) stain, to obtain precise information concerning the size, shape and color of cellular elements. A number of factors are taken into consideration including the zone of microscopic observation, the quality of staining, condition of grabbing images and the size of the sample. Morphometric and colorimetric measurements have been applied as objective tools for demonstrating the predominance of each Red/Green/Blue component, and more particularly their reciprocal ratio. The results obtained for each center may constitute a guideline for adjusting their procedure, in order to reach a consensus regarding similar staining by all centers.

Azure Stains↗
PubMed1979

Paraformaldehyde-induced fluorescence as a histochemical test for 5-hydroxytryptamine in the epididymis of the opossum.

Epididymides of four opossums were tested histochemically for the presence of 5-hydroxytryptamine (5-HT). In addition, testes and epididymides of two opossums were analysed biochemically for the same substance. Biochemical analysis revealed a high concentration of 5-HT in the epididymis, while the amount in the testis proved to be insignificant. Fluorescence microscopy gave positive results in the apical portion of the epithelium. There was also fluorescence in the connective tissue in general, in mast cells, and in a thick subepithelial band in particular. Following treatment with 2% sodium borohydride in 1,2-propanediol for 15 minutes, the fluorescence of mast cells was removed, while there was a slight reduction in epithelial fluorescence and that in the subepithelial band and other connective tissue. The subepithelial band stained metachromatically with azure B, and this staining characteristic was removed by treatment with RNAse. Feulgen reaction revealed elongate nuclei in the band. It would seem that the 5-HT is located in both the mast cells and the epithelium. It seemed to be also present in the subepithelial band, but because of strong autofluorescence it is difficult to demonstrate. At the present time the function of 5-HT in the opossum epididymis is unknown, and is under further investigation.

Animals↗
PubMed1982

Demonstration of chlamydial inclusions in exfoliated cells.

Microscopic demonstration of chlamydial inclusions within cells offered the first laboratory procedure supporting the clinical diagnosis of chlamydial infection. The cytological test for the detection of chlamydial inclusions in genital tract infection, through not as sensitive as isolation in cell cultures, is still of diagnostic value. The present study discusses the collection of clinical specimens for microscopic examination, the preparation, fixation and staining of slides by Giemsa stain, iodine, and immunofluorescence.

Adult↗
PubMed1984

[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].

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

Animals↗