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PubMed1984

Detection of experimental Chlamydia trachomatis eye infection in conjunctival smears and in tissue culture by use of fluorescein-conjugated monoclonal antibody.

The immunofluorescent staining of conjunctival cytology smears with an antichlamydial monoclonal antibody is a new method for the detection of chlamydial eye infection. Using a monkey model of chlamydial eye infection, we compared the monoclonal antibody staining cytology with the established methods of Giemsa-stained cytology and chlamydial culture with cycloheximide-treated McCoy cells. Compared with either culture or monoclonal antibody-stained cytology, Giemsa cytology had a low sensitivity (30 and 24%, respectively) but was highly specific (100% in each case). Compared with culture, monoclonal antibody-stained cytology was highly sensitive (94%) but appeared to lack specificity (70%). However, examination of the changes in identification rates with time suggested that monoclonal antibody-stained cytology may be even more sensitive than culture and detect lower levels of infection. This study showed that monoclonal antibody-stained cytology of conjunctival smears might be a rapid, efficient, and inexpensive method of diagnosing ocular chlamydial infection.

Animals↗
PubMed1977

Banded chromosomes of the owl monkey, Aotus trivirgatus.

Chromosomes of the owl monkey, Aotus trivirgatus, with 2n=54, 53, or 52, have been stained to show quinacrine (Q-) and Giemsa (G-) bands, and a karyotypic arrangement has been proposed based on lengths, centrometric index, and banding pattern. C-bands were present at the centromeric region of every chromosome and over the entire short arm of certain acrocentric chromosomes; 5-methylcytosine was concentrated in the same regions. Bright Q-bands at the telomeric ends of the short arms of some chromosomes probably represent a second type of repetitive DNA. Ag-staining showed that only the chromosomes bearing a secondary constriction are nucleolus organizer chromosomes.

Animals↗
PubMed1990

Utility of Gram's and Giemsa stains in the diagnosis of pulmonary tuberculosis.

Conflicting information in the literature regarding the staining properties of Mycobacterium tuberculosis using Gram's stain and experience in two patients with pulmonary tuberculosis in whom the diagnosis was suspected after staining with non-acid-fast bacillus stains prompted the study of Gram's stain in this disease. The main finding was that mycobacteria appear as refractile, gram-neutral, or faintly gram-positive bacilli after Gram's stain, depending upon the plane of focus in which the organisms are regarded. It is concluded that the diagnosis of mycobacterial disease may be suggested by Giemsa- and Gram-stained smears of clinical specimens. M. tuberculosis may be either gram-neutral or gram-positive.

Adult↗
PubMed1999

Cryptosporidium infection among sheep in Qalubia Governorate, Egypt.

The present study was carried to explore the incidence of Cryptosporidium infection among lambs in Qalubia governorate as well as to study the ability of three staining techniques developed for rapid identification of Cryptosporidium oocysts in faecal samples of 120 neonatal lambs less than one month old. The stains used in this investigation were modified Zeihl-Neelsen stain (MZN), Safranin-methylene blue (SMB) and Giemsa stain (GS) to detect the recovered oocysts with their incidence variation. The diagnostic value was raised by direct smear and concentration floatation technique. The infection rates with Cryptosporidium oocysts among the investigated faecal samples from neonatal sheep were 82, 58, 36, out of 120 (68.3, 48.3 and 30%) by modified Zeihl-Neelsen stain, Safranin-methylene blue and Giemsa stain respectively. The lambs less than one week had higher infection rate. The relationship between age susceptibility and infection was discussed. The use of modified Zeihl-Neelsen stain was currently recommended procedure with the concentration floatation technique for the accurate diagnosis and rapid identification of Cryptosporidium oocysts.

Animals↗
PubMed2006

[DNA amplification of Plasmodium vivax parasites from Giemsa-stained blood smears].

OBJECTIVE: To develop methods of extracting DNA from malaria parasites on Giemsa-stained blood smears. METHODS: Improved Na2HPO4 method and Chelex-100 ion-exchange technique were used to extract DNA from Giemsa-stained or unstained blood smears. Nested PCR was employed for amplification and identification of allelotype in the Plasmodium vivax merozoite surface protein-1 (PvMSP-1). RESULTS: Target DNA bands appeared in all samples of unstained thick blood smears, while no DNA bands were visible in the fixed and stained thin smears. Both methods identified PvMSP-1 alleles from smears with parasitemia of > or = 0.01%. CONCLUSION: It is feasible to identify PvMSP-1 alleles from Giemsa-stained blood smear.

Alleles↗
PubMed1982

Polymorphisms of Ag-stained nucleolar organizer regions in man.

Polymorphisms of the NORs as tested by Ag-staining of metaphase G-banded chromosomes were investigated in cultured blood lymphocytes of karyotypically normal individuals from the Moscow population. The study of cell-to-cell variability in the number of Ag-stained NORs carried out on 14 monozygotic twin pairs showed the phenomenon to have some features of real intercellular variation. In 40 unrelated individuals the individual acrocentric chromosomes were compared by the number of Ag-stained NORs, their degree of staining, and their participation in acrocentric association. Chromosome 21 was found to be significantly more active than four others by all the criteria, and chromosome 15 was less active compared with the others by the size of the Ag deposits and the frequency of participation in NOR associations. The frequency distribution of homozygotes and heterozygotes for Ag-stained NORs in the same group of 40 individuals was in accordance with the Hardy-Weinberg law.

Azure Stains↗
PubMed1996

Increase in epithelial mast cell numbers in the nasal mucosa of patients with perennial allergic rhinitis.

The aim of the study was to compare the numbers and distribution of mast cells in the nasal mucosa of perennial allergic rhinitis (PAR) patients and controls, as demonstrated by different staining methods for light microscopy. Biopsies of inferior turbinate mucosa were taken from 10 patients with PAR and 10 patients undergoing septoplasty or septorhinoplasty (control group). Sections for light microscopy were stained with azure A. chloroacetate esterase and an ABC immunohistochemical technique using antibody to tryptase. Three times more mast cells were found in the epithelium of PAR patients compared to controls using the immunohistochemical technique (p = 0.0074). This method demonstrated considerably more mast cells than the other stains. The increase in epithelial mast cells is consistent with the migration of mast cells seen in seasonal allergic rhinitis, and this may be important in the phenomenon of nasal priming seen after repeated antigen exposure.

Cell Count↗
PubMed1998

Effect of prolonged staining with hematoxylin on detecting Helicobacter pylori in hematoxylin-eosin-stained gastric mucosa.

Helicobacter pylori (H. pylori) is identifiable in hematoxylin-eosin (HE)-stained preparations, particularly under a prolonged hematoxylin staining condition. The staining intensity of H. pylori bodies was increased when the hematoxylin staining period was doubled from 5 min, the routine period of Meyer's hematoxylin staining, to 10 min. The staining intensity of the background epithelial nuclei was only mildly increased. Hematoxylin-eosin staining employing a prolonged hematoxylin staining period gave detectability of H. pylori comparable to May-Giemsa and immunoperoxidase staining. The modified HE method is thus very useful and practical for identifying H. pylori in routine gastric biopsy specimens.

Adult↗
PubMed1991

Detection of Plasmodium falciparum infection with the fluorescent dye, benzothiocarboxypurine.

The fluorescent dye benzothiocarboxypurine (BCP) intensely stains nucleic acids. The dye does not penetrate viable white blood cells but does stain these cells following fixation. It has also been found that the dye stains the nucleic acid of viable Plasmodium falciparum. We have subsequently evaluated the staining of P. falciparum by benzothiocarboxypurine within red blood cells and have found that the red blood cell membrane is freely permeable to this dye and consequently P. falciparum is stained within the red blood cell. This finding prompted an in-depth analysis of the dye in the laboratory and in a field study as an alternative to Giemsa-stained blood smears and as a means of enhancing the microscopic diagnosis of malarial infection. In a field study the BCP dye allowed detection of malaria in fresh blood at a level equivalent to the Giemsa method (parasitemia ranged from 0.01% to 30%). The BCP staining procedure could also be used with fixed specimens although the differential staining characteristics were lost following specimen preparation. Of 111 blinded samples obtained in the field 22 were negative by Giemsa-stained thin smear, 16 were negative on thick smear and the same 16 were negative by BCP analysis. We have found that the BCP dye offers many advantages compared with the microscopic diagnosis of P. falciparum infection with standard Giemsa stains. These advantages are especially evident in conditions of low parasitemia, in the speed of staining and evaluation, and the relatively low level of training required to provide consistent results.

Animals↗
PubMed2000

'Glowing' chromosomes in cells undergoing rapid division.

Previous investigations in which metaphase plates of cells in rapid division were incubated in phosphate buffer at high temperature revealed numerous heterochromatic dots in chromosomes after Giemsa staining. In contrast, chromosomes from cells with a reduced capacity for reproduction were devoid of such dots, or the dots were sloughed-off into rings and patches of heterochromatin. In two types of cells which were rapidly dividing, namely HeLa cells (cervical cancer) and cells from regenerating planaria, phosphate incubation followed by Giemsa staining revealed an 'aura' or 'glowing' effect on the chromosomes, consisting of a densely staining core surrounded by a lightly stained periphery. This finding might be developed into a diagnostic test for certain malignancies, for cells undergoing dedifferentiation, or for tissues undergoing regeneration.

Animals↗
PubMed1988

Pneumocystis carinii in sputum. Comparable efficacy of screening stains and determination of cyst density.

To determine whether accurate quantification of Pneumocystis carinii cysts in sputum is possible using currently available means and to determine the optimal stain for screening sputum for P carinii, we undertook a comparative evaluation of readily available stains: (1) Diff-Quik, a stain that provides tinctorial detail comparable with Giemsa, (2) toluidine blue, and (3) silver methenamine on simultaneously thawed sputum samples containing P carinii. We also studied the Diff-Quik stain in combination with (4) toluidine blue and (5) silver methenamine. The three stains were comparable for screening purposes. Each disclosed similar numbers of P carinii aggregates on similarly prepared slides. Cysts were difficult to recognize on Diff-Quik due to the negatively staining character of the cyst wall with this stain. The cyst-wall stains, toluidine blue and silver methenamine, clearly stained the cyst wall. The combination of a cyst-wall stain with Diff-Quik allowed visualization of cysts, highlighted by the cyst-wall stain, and aggregate area, delineated by the Diff-Quik, allowing expression of number of cysts per aggregate area, a measure termed cyst density.

Animals↗
PubMedDate not supplied

[Comparison of 2 dyes in the detection of Pneumocystis carinii].

OBJECTIVE: To evaluate two staining procedures used in the detection of P. carinii. MATERIAL AND METHODS: Bronchoalveolar lavage samples were obtained from 20 immunosupressed rats. The samples were stained by three procedures: o-toluidine blue (OTB), Diff Quik (DQ), and a silver stain as the gold standard. The microscopic search for P. carinii was made independently by two observers. RESULTS: Fifteen of the 20 samples were positive to P. carinii with the silver stain. The sensitivity with DQ was 27% and the specificity 100%. With OTB the sensitivity was 93% and the specificity was 80%. The inter-raters' agreement by kappa was 0.11 for DQ and 0.53 for OTB. CONCLUSION: Our results suggest that the silver stain should be used for an adequate diagnosis of pneumonia caused by P. carinii.

Animals↗
PubMed1979

A comparison of constitutive heterochromatin staining methods in two cases of familial heterochromatin deficiencies.

Using DAPI staining after pretreatment with distamycin A we detected a familial deficiency of chromosome 16 heterochromatin. A distinct positively staining band, however, was seen after C-banding. Thus, by using these different heterochromatin staining methods, heterogeneity of the constitutive heterochromatin in the centromeric region of human chromosome 16 was indicated. The same C-banding procedure was also applied to a previously described familial deficiency of chromosome 9 heterochromatin evidenced using distamycin A/DAPI staining and G 11 staining (Buys et al., 1979). In this case a C-band appeared to be virtually absent on the relevant chromosome. These staining methods may be valuable tools in the study of chromosome polymorphisms.

Azure Stains↗
PubMedDate not supplied

Understanding microwave-stimulated Romanowsky--Giemsa staining of plastic embedded bone marrow.

Bone marrow smears were made and fixed in methanol or formaldehyde. Marrow sections of various thicknesses were also prepared from formaldehyde fixed marrows embedded in paraffin or plastic (glycol methacrylate). The different smears and sections were then stained by a Romanowsky--Giemsa procedure. Some specimens were stained using a standard microwave-stimulated method previously used diagnostically. The effects of technical variations were studied, including degree of microwave irradiation and the staining time. Comparisons of the resulting staining outcomes showed that microwave stimulated Romanowsky--Giemsa staining of plastic sections is a rate controlled process. Unusual aspects of the staining pattern of plastic sections (namely the purple basophilic cytoplasms and nucleoli, and blue chromatin) are due to microwave stimulation and formaldehyde fixation respectively.

Animals↗
PubMed2002

Comparison of immunohistochemical and modified Giemsa stains for demonstration of Helicobacter pylori infection in an African population.

BACKGROUND: Modified Giemsa staining has been favoured by many researchers because it is easy to perform but, like many other stains, demonstration of the bacteria depends on its morphology. It has been arged in some research circles that some of the organisms in the gastric mucosa may not be true H.pylori. Immunohistochemical techniques have been developed and make use of anti H.pylori antibody, which reacts, with somatic antigens of the whole bacteria and have been found to correlate well with the presence of the bacteria. OBJECTIVE: To ascertain the efficacy of modified Giemsa stain in an African setting where H.Pylori seems quite prevalent. STUDY DESIGN: A laboratory-based study of two diagnostic tests in which modified Giema stain was compared with immunohistochemistry. METHODS: A total of 48 consecutive autopsy cases with no upper gastro intestinal diseases had their gastric mucosa stained for demonstration of H.pylori using both modified Giemsa and immunohisto chemical staining techniques. RESULTS: Twenty-seven cases of H.pylori were demonstrated by both techniques and 14 cases were not identified by the two staining methods. In 2 cases immunostain could not demonstrate the bacteria but they were identified with modified Giemsa stain while in 5 cases the bacteria were identified by immunostain but not with modified Giemsa stain. The sensitivity of modified Giemsa stain was 85% (CI 66.5-98.8) while the specificity was 89% (CI 60.4 - 97.8). The positive predictive value of modified Giemsa stain was 93% CI 75 - 98.8%) while the negative predictive value was 74% (CI 48.6 - 89.9). The kappa statistic comparing the 2 stains was 0.69 (p value 0.00001) giving a good agreement between the two tests. CONCLUSION: With the above results the modified Giemsa stain, which is readily available in most African laboratories, is recommenced for diagnosis of H.pylori, a prevalent infection in Africa.

Adolescent↗
PubMed1995

Determination of the test performance of less costly methods of Helicobacter pylori detection.

Since Helicobacter pylori (Hp) has become so important in the management of peptic ulcers, it is imperative that noninvasive and inexpensive methods of diagnosis be available. The objective of this prospective, double-blind study was to determine the performance of the noninvasive 14C-urea breath test and the less expensive hematoxylin-phloxin-saffron (HPS) and Giemsa stains in the detection of this infection. As well, we set out to determine the minimal number of biopsies required to make an accurate diagnosis. Fifty consenting adults referred for gastroscopy underwent antral biopsies for HPS, Giemsa and Steiner silver staining and a 14C-urea breath test. The ability of HPS, Giemsa, and the breath test to differentiate between the presence or absence of Hp infection as defined by the silver stain was assessed using contingency table analysis. Hp was detected in 24 patients with the silver stain. Half of these patients had evidence of ulcer disease, compared to 3 of 26 patients who were negative for Hp on silver stain (p < 0.01). The Hp positive group were more frequently male (62% vs. 31%, p < 0.05) and more often had chronic active gastritis (96% vs. 7.7%, p < 0.001). The sensitivity of the breath test, HPS, and Giemsa stains were 95.8% (95% confidence interval: 79-100%), 75% (53-90%), and 95.8% (79-100%), respectively. Specificity was 100% (87-100%) for all 3 methods of detection. The breath test was also able to discriminate between heavily infected and moderately infected patients as defined by silver stain (p < 0.05), and could be shortened from 30 to 15 min without any loss of accuracy.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗
PubMed1995

Serosal balls detected immunocytochemically in peritoneal lavage obtained during surgery.

Using the immunoperoxidase technique, we studied "serosal balls," which have features resembling those of cells from primary and metastatic tumors, and may thus complicate cytodiagnosis. Serosal balls were detected in 32 (18%) of 174 peritoneal washings. The balls consisted of oval clusters of cells in solid masses surrounded by flattened cells. The interior of the serosal balls was stained green with Papanicolaou method, showing the presence of homogeneous amorphous material, sometimes stained in a filamentous pattern. Almost all serosal balls were stained immunocytochemically for both keratin and vimentin. The interior was stained with antibodies against collagen types I and III. Therefore, these balls were fragments of serous membrane, and contained fibrous tissue and mesothelial cells.

Azure Stains↗
PubMed1981

DNA extraction during Giemsa differentiation of chromatids singly and doubly substituted with BrdU.

Human lymphocytes were cultured in 3H-labelled BrdU. Cells were pretreated to induce differentiation, autoradiographed and Giemsa-stained. DNA extraction was deduced if grain counts were lower in differentiated mitoses compared with untreated controls.--The differentiation method involved sequential pretreatments with short wave UV and 2 x SSc at 60 degrees C. This removed 34% of label from first division cells (with TB.TB chromosomes) but relatively more (53%) from second division (TB.BB chromosomes). In second division cells, about two thirds of label was lost from pale (BB) chromatids but only one third from dark (TB) chromatids. The UV and SSC pretreatments acted in collaboration, since neither alone reduced grain counts significantly.--On testing other methods, similar preferential DNA extraction was obtained with Perry and Wolff's FPG method, and with the hot salt pretreatment of Korenberg and Freedlender. However, good Giemsa differentiation could also be obtained using Hoechst 33258 and light pretreatments without any DNA loss. Reverse differentiation patterns (TB pale, BB dark) induced by warm acids resulted in extraction of nearly two thirds of 3H-BrdU label, but relative loss was the same from pale and dark chromatin. Direct reverse staining using alkaline Giemsa did not result in any loss of label.--Thus preferential DNA loss from pale stained chromatin underlies differentiation methods using light plus hot salt pretreatments, but it is not obligatory for good differentiation using other techniques.

Autoradiography↗