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At least 217 records · Page 12Linked to original sources
PubMed1997

Application of the RB-FPG technique for distinguishing early and late replicating X chromosomes in farm animals.

The sex chromosomes of Murrah buffalo and Sahiwal cattle (females) were studied for R-banding patterns. A short term whole blood culture method was used for obtaining metaphase chromosomes. The R-banding method using 5-bromo-2'-deoxyuridine (BrdU), the fluorescent 33258 Hoechst stain followed by blue-black light exposure and Giemsa staining (RB-FPG) was applied to the chromosome preparations. The two X chromosomes in all the females were clearly distinguished. One of the chromosomes (early replicating) had alternative light and dark bands and the other X chromosome (late replicating) was either completely pale banded or had one or a few dark bands. The reason for this phenomenon could be BrdU incorporation in the late replicating X chromosome, which on exposure to blue light in the presence of 33258 Hoechst is destroyed and does not take the stain. Thus, one of the two X chromosomes shows differential staining behaviour.

Animals↗
PubMed1993

Acute conjunctivitis in childhood.

We obtained specimens for culture from the lids and conjunctivae of 95 patients with acute conjunctivitis and 91 control children of similar age and, in addition, stained the conjunctival scrapings with Giemsa and Gram stains. The conjunctivitis was attributed to bacterial infection in 76 patients, viral infection in 12 children, and allergy in 2 patients; no cause was identified in the remaining 5 patients. In most cases the etiologic diagnosis was based on the results of laboratory studies. By separately culturing microorganisms in specimens from the lids and conjunctivae of patients and control subjects, we could distinguish normal flora from pathogens, and blepharitis from conjunctivitis. Staphylococci, corynebacteria, and alpha-hemolytic streptococci were the predominant organisms recovered from the lids of control subjects. In contrast, Haemophilus influenzae, Streptococcus pneumoniae, and Moraxella catarrhalis were the major pathogens cultured from the conjunctival specimens from patients with bacterial conjunctivitis. Gram stains of conjunctival scrapings provided a rapid means of predicting the pathogen in 51 of 55 cases of bacterial conjunctivitis. Giemsa stains of conjunctival scrapings provided etiologic information in 81 of 84 cases, showing neutrophilia in bacterial infections, lymphocytosis in viral infections, and eosinophilia in allergic disease. These results indicate that most cases of acute conjunctivitis in children can be diagnosed on the basis of differential cultures of microorganisms from the lid and conjunctiva, together with Giemsa stains of conjunctival scrapings.

Acute Disease↗
PubMed1980

A comparison of direct fluorescent antibody and Giemsa staining for the post-mortem diagnosis of anaplasmosis.

Direct fluorescent antibody (DFA) and Giemsa staining of Anaplasma marginale were compared in smears collected serially at post-mortem (PM) from 11 experimentally infected calves. Once smears had been prepared and air-dried they could be held for at least 5 days before staining with either technique with no noticeable change in staining quality. DFA staining was more sensitive in detecting anaplasms in smears than Giemsa staining. Anaplasma spp could be differentiated from Babesia bovis and B. bigemina by DFA staining but there were cross reactions between A. marginale and A. centrale. Blood smears prepared from subcutaneous vessels in the legs provided better diagnostic material than kidney, heart and lung smears. Brain smears were not suitable for PM diagnosis using either staining technique.

Anaplasmosis↗
PubMed1980

A comparison of the sensitivity of immunofluorescence and Giemsa for staining Chlamydia trachomatis inclusions in cycloheximide-treated McCoy cells.

In a clinical study of 190 men with non-gonococcal urethritis, Chlamydia trachomatis inclusions were sought in cycloheximide-treated McCoy cells by an indirect immunofluorescent antibody technique. The method was consistently reliable over a period of two years, and the results were obtained within 24 hours of a patient's attendance. The results correlated with those obtained by Giemsa staining in 91.6% of patients, and the new method was at least as sensitive as the established Giemsa-staining method.

Azure Stains↗
PubMed1981

The staining of the synaptonemal complex for light microscopic study in the mouse.

Demonstration of the synaptonemal complex for light microscopy has until now been based on staining with silver. After fixation at pH 9-10 it is also possible to visualize synaptonemal complexes with several nonspecific protein stains such as Coomassie brilliant blue, Giemsa, fast green, light green and Stains All. Although staining with silver gives the best contrast between synaptonemal complexes and the background, the other dyes have a number of advantages, such as more even staining, easy extractability, and lower cost than silver.

Animals↗
PubMed1979

Autopsy diagnosis of Legionnaires' disease in immunosuppressed patients. A paleodiagnosis using Giemsa stain (Wohlbach modification).

A diffuse pneumonia characterized by short Giemsa-stained, gram-negative rods that were not cultured was diagnosed at autopsy in the fall of 1975 in three immunosuppressed patients who had had renal transplants. An acute fibrinous exudate was found, with macrophages, neutrophils, and numerous short bacilli in each inflamed alveolus as shown by Giemsa's stain (Wohlbach modification). Transplantation was stopped for 1 month. The institutional infection control committee discussed the cases, and increased observation. After the Legionnaires' disease agent was identified in 1977, the bacteria were stained by Dieterle's method and identified as Legionnaires' disease bacteria by direct fluorescent antibody test. No further cases have been identified in the succeeding 62 renal transplantations at this medical center. Although Wohlbach's modification of Giemsa has given consistently good staining of the Legionnaires' disease bacterium in our cases, unclarified factors may interfere with good staining, and the May-Grünwald Giemsa stain was unsatisfactory.

Adult↗
PubMed1982

On the mechanism of differential Giemsa staining of BrdU-substituted chromatids.

This paper analyses the effect of acid hydrolysis on the differential Giemsa staining of 5-bromo-2'deoxyuridine (BrdU) substituted chromatids in human and plant chromosomes, after treatment with a fluorochrome and light. Human lymphocytes and Allium cepa L. root tips were grown in BrdU for two or three cell cycles. Lymphocytes spreadings and meristem squashes were treated with fluorochrome Hoechst 33258, exposed to sunlight, hydrolysed with 5N HCl and stained with giemsa. This acid hydrolysis improves the differential staining of BrdU substituted and non-substituted chromatin. It also allows the differentiation of sister chromatids with the DNA specific dye azure-A.

Azure Stains↗
PubMed2005

Ultrarapid staining for cutaneous melanoma: study and protocol.

BACKGROUND: Immunohistochemistry has revolutionized the use of frozen sections for the removal of melanoma. OBJECTIVE: Two studies were performed to address the best and fastest way to stain melanocytes in frozen tissue sections. MATERIALS AND METHODS: Fifty-six standing cones were taken from sun-damaged white skin and stained with combinations of anti-S-100, anti-HMB-45, anti-Melan-A, and azure B with appropriate controls. RESULTS: Anti-Melan-A was the best immunohistochemical stain. Azure B stains melanin granules metachromatically green and allows differentiation of melanocytes and melanophages. Ultrarapid staining worked as well as rapid staining. CONCLUSION: Dako's EnVision ultrarapid staining protocol and azure B counterstain represent a dramatic improvement and should be considered the easily obtainable histologic standard when excising cutaneous melanoma.

Frozen Sections↗
PubMed1999

Retinal pathology of canine X-linked progressive retinal atrophy, the locus homologue of RP3.

PURPOSE: To describe the course of photoreceptor disease in canine X-linked retinal degeneration. METHODS: Retinas from 55 dogs (44 males, 8 carrier females, 3 homozygous females) were obtained by enucleation under general anesthesia. After fixation and dehydration, tissues were embedded in epoxy resin, sectioned at 1 microm for light microscopy and stained with azure II/methylene blue and a paraphenylenediamine counterstain. For electron microscopy, regions identified by light microscopy were selected and cut at 60 nm. Sections were stained with uranyl acetate-lead citrate. Electroretinography from an additional group of normal males, affected males, and carrier females was performed and the rod and cone responses evaluated. RESULTS: The earliest lesion detectable by electron microscopy was vesiculation of rod discs, followed by disruption of outer segments and death of rods. Loss of cones and progressive atrophy of inner retinal layers followed. Lesions were most severe in the peripheral retina and advanced toward the optic disc with disease progression. Significant variation in disease severity was present in males despite the presence of the same disease allele in all affected dogs. Carrier females displayed generalized reduction in photoreceptor density as well as multifocal areas of complete rod loss. The electroretinogram (ERG) findings were compatible with the histopathologic abnormalities. Homozygous females had lesions similar to those seen in affected males. CONCLUSIONS: X-linked retinal degeneration is characterized by initial degeneration of rod photoreceptors, followed by loss of cones and progressive atrophy of the inner retina. Carrier females display a phenotype consistent with random X-chromosome inactivation. Variation in genetic background may alter expression of the disease allele in affected animals, thus accounting for variation in phenotypic expression of the disease.

Animals↗
PubMed1980

In vivo assay of viability of amastigotes of Leishmania donovani.

Following intravenous (i.v.) injection of formalin-killed or ultraviolet-killed or ultraviolet-irradiated amastigotes of Leishmania donovani, the parasites could be identified in mouse liver at 5 min, but most amastigotes were digested by 3 hr or between 24 and 72 hr, respectively. An in vivo assay of viability, based on these observations, suggests differences in the viability of hamster amastigote populations. The identification of dead and/or dying amastigotes in the liver 24 hr after i.v. injection suggests that enumeration of amastigotes from Giemsa-stained slide preparations may include varying numbers of nonviable organisms.

Animals↗