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At least 289 records · Page 16Linked to original sources
PubMed1998

Spectrally resolved morphometry of the nucleus in hepatocytes stained by four histological methods.

A novel concept of spectrally resolved morphometry for histological specimens was developed using light microscopy combined with spectrally resolved imaging. The spectroscopic characteristics of rat hepatocytes stained by Haematoxylin and Eosin, Romanowsky-Giemsa, periodic acid-Schiff and Masson's trichrome were assessed. Light intensity in the range 450-850 nm was recorded from 10000 pixels of nuclear domains of each stained cell and represented as light transmittance spectra and optical density. In order to identify spectral shifts caused by stain-macromolecule interactions, we compared the spectra of individual stain components with those of DNA and bovine serum albumin. Chromatin and interchromatin areas were classified spectrally using a chosen spectral library followed by morphometric calculations of nuclear domains for each staining method. The spectral fingerprints of Masson's trichrome stain distinguished the nucleolus from the rest of the nuclear chromatin, enabling the demarcation and calculation of the nucleolar area. Spectrally resolved imaging of human hepatocytes stained by Masson's trichrome stain revealed marked differences between the nucleolar area in normal human hepatocytes compared with hepatocellular carcinoma. Masson's trichrome stain also distinguished the nucleolar area in human breast carcinoma cells and keratinocytes.

Animals↗
PubMed1987

A rapid modified fluorescence-plus-Giemsa technique for sister chromatid exchanges and DNA replication patterns.

Five different fluorochromes were tested, in a revised fluorescence-plus-Giemsa technique, for their ability to give good quality permanent preparations. Hoechst 33342 was found to give the best results. Slides were stained with Hoechst 33342, followed by exposure to long wavelength UV light with a simultaneous incubation in a hot alkaline solution, and then were Giemsa stained. This technique gives good quality preparations, in approximately 1 h, of sister chromatid exchanges or DNA replication patterns.

Azure Stains↗
PubMed1988

Monoclonal antibodies in the laboratory diagnosis of trachoma.

Various techniques which use monoclonal antibodies to detect Chlamydia trachomatis in clinical specimens are reviewed. An investigation comparing the efficacy of immunofluorescent staining with Giemsa staining in detecting Chlamydia in conjunctival scrapings from cases of active trachoma is presented. Sixty-two eyes of schoolboys with moderate to severe trachoma were studied. Giemsa staining detected chlamydial inclusion bodies in 34 percent of the specimens. Free elementary bodies were detected by fluorescent monoclonal antibody in 21 percent. Eleven percent were positive by both Giemsa and immunofluorescence and 55 percent were positive by either Giemsa and/or immunofluorescence. The addition of fluorescent monoclonal antibody assay to routine Giemsa staining resulted in an increase in the yield of positive specimens by 29 percent.

Antibodies, Monoclonal↗
PubMed1990

A metachromatic dye-ATPase method for the simultaneous identification of skeletal muscle fiber types I, IIA, IIB and IIC.

The histochemical demonstration of quantitative differences in myofibrillar ATPase activity at the selective pH optima of the various types of human skeletal muscle fibers is the most widely used technique for their differentiation. The basis of the reaction is the deposition of insoluble salts of inorganic phosphate cleaved from ATP by myofibrillar ATPase(s) followed by substitution of the phosphates with less soluble chromogenic salts. Doriguzzi and associates reported using metachromatic dyes to demonstrate quantitative differences in phosphate deposition among different fiber types. Following routine ATPase histochemistry and staining with either azure A or toluidine blue, fibers with low ATPase activity (and low phosphate content) were stained metachromatically while fibers with high ATPase activity (and high phosphate content) were orthochromatic with the intensity of color proportional to the content of insoluble phosphate. The metachromasia was readily lost after immoderate washing in aqueous solutions or routine dehydration in ethanol, with consequent diminished fiber type distinction. A critical modification of this technique is reported in which incubation of frozen sections of human skeletal muscle in ATP-containing medium is carried out at room temperature (22-24 C), rather than the usual 37 C, followed by a revised washing and dehydration protocol. With these modifications, the four human skeletal muscle fiber types (types I, IIA, IIB, and IIC) can be identified rapidly and reliably in single sections, obviating the need for examination of serial sections. The tinctorial differentiation allows fiber type identification even in black and white photographs.

Adenosine Triphosphatases↗
PubMed1996

The Ag-NOR proteins present a crescent-shaped distribution at the secondary constrictions of metaphase PtK1 chromosomes.

The ultrastructural distribution of the proteins responsible for the silver staining of NORs was analyzed on serial thin sections of preembedded silver-stained rat kangaroo PtK1 metaphase chromosomes. Our results show that the Ag-NOR proteins present a crescent-shaped distribution at the secondary constriction of each chromatid. Moreover, in some cases the crescent-shaped structures of both chromatids are not symmetrically arranged but show different orientations. These observations, together with our own and previously reported light microscopy results obtained on silver-stained mammalian chromosomes, lead us to suggest a mechanism for the formation of apparent secondary constrictions at NORs.

Animals↗
PubMed2004

Demonstration of different endocervical staining methods and their usefulness in the diagnosis of the chlamydial infection in exfoliated cells advantages and disadvantages.

Microscopic demonstration of chlamydial inclusions within cells offered the first laboratory procedure supporting the clinical diagnosis of chlamydial infection. Our aim is to evaluate the usefulness of different endocervical staining methods in diagnosis of Chlamydia trachomatis (CT) infection within exfoliated cells of the endocervix. The cytological test for the detection of chlamydial inclusions in genital tract infection, though not as sensitive and specific as isolation in the cell culture monolayers, is still of the diagnostic value. The present study discusses the collection of clinical smears for microscopic examination, their preparation; fixation and staining of slides by a variety of staining methods that have been used to detect Chlamydia in clinical smears and biopsies. Most of these methods such as Giemsa stain, Papanicolaou, iodine, and immunofluorescence (IF) using monoclonal antibodies, are based on the combination of dyes designed to obtain optimum differentiation of the various structures. The utilization of different endocervical smear stains together with the clinical information can be used to identify women at high risk for CT infection.

Azure Stains↗
PubMedDate not supplied

Diagnosis of herpes simplex virus-1 keratitis: comparison of Giemsa stain, immunofluorescence assay and polymerase chain reaction.

PURPOSE: To evaluate three different diagnostic tests against the gold standard of viral isolation, in the diagnosis of HSV-1 keratitis. METHODS: Corneal scrapings from 170 patients with clinically suspected HSV keratitis were tested by; 1) Giemsa staining procedure for the presence of multinucleated giant cells and lymphocytes, 2) immunofluorescence assay for HSV-1 antigen, 3) polymerase chain reaction (PCR) for HSV-1 DNA and 4) virus isolation by shell vial culture in SIRC (Rabbit corneal epithelial cell line). The results of the former three tests were compared among 14 cases that were culture positive and 156 cases that were culture negative for HSV-1. RESULTS: The sensitivity of PCR was 100%, while IFA and Giemsa had sensitivities of 85.7% and 57.1% respectively. The specificity of PCR, IFA and Giemsa were found to be 67.9%, 85.3% and 85.9% respectively. CONCLUSIONS: In the present study, a combination of PCR and immunofluorescence assay appears to be the most suitable choice of tests for diagnosis of HSV-1 keratitis, while detection of MNGC by Giemsa staining procedure may give us a presumptive diagnosis of suspected viral infection.

Azure Stains↗
PubMed1990

[Measurement of the DNA content in morphologically identified cells on a Wright-Giemsa stained smear].

4',6-Diamidino-2-phenylindole (DAPI) staining was applied to measuring nuclear DNA contents of single cells previously identified on a Wright-Giemsa stained smear. The smear was first photographed and the location of each cell was mapped. The smear was then destained with 50% acid ethanol and absolute methanol, and restained with DAPI (pH 7.4) at 4 degrees C. Blue fluorescence was observed in the nucleus, and the intensity of this fluorescence was proportional to the amount of DNA, with the lowest coefficient of variation of 3.6% in the case of staining the peripheral lymphocytes for 30 min. This method is useful for measuring DNA contents in heterogeneous cell populations.

Azure Stains↗
PubMedDate not supplied

The alcohol-fixed Diff-Quik stain. A novel rapid stain for the immediate interpretation of fine needle aspiration specimens.

A rapid alcohol-fixed Diff-Quik stain is described. The technique takes less than five minutes and results in excellent cytologic detail, comparable to that with Papanicolaou stain, as well as superb stromal definition. The stain should be very useful to pathologists and cytotechnologists for immediate interpretation of fine needle aspiration specimens.

Azure Stains↗
PubMed1995

Detection of Theileria parva in the salivary glands of Rhipicephalus appendiculatus: evaluation of staining methods.

A comparison of ten methods for staining tick salivary glands for detection of Theileria parva infection from ticks fed on rabbits for various periods was undertaken. Staining with azure without hydrochloric acid hydrolysis was found to be the most reliable method for detection of the presporozoite stages (sporoblasts) of T. parva in the salivary gland acini of unfed Rhipicephalus appendiculatus and could be applied to field ticks. All the stains proved suitable for the detection and quantitation of sporozoites in ticks fed for 4 days on rabbits. The capacity of the stains to allow detection of early stages of T. parva differed, but it became more reliable during tick feeding as sporoblasts developed and matured. Giemsa's stain and Feulgen's stain followed by superimposition of Giemsa's stain were superior to other stains for the detection and quantitation of immature salivary gland stages in feeding ticks.

Animals↗
PubMed1978

The characterization of high-resolution G-banded chromosomes of man.

The detailed characterization of G-banding patterns of high resolution human chromosomes has been possible with the utilization of a refined cell synchronization technique which routinely yields a large number of excellent quality cells in late prophase, prometaphase, early metaphase, and mid-metaphase. The mitotic cells exhibit up to a 400% increase in the number of bands previously visualized by standard methods. From studies of the banding patterns, it has become evident that the G-positive and, to some extent, the G-negative bands of mid-metaphase results from a coalescence of finer subbands of earlier stages and that each band and its corresponding subbands maintain a constant location throughout the process of chromosome condensation. A precise schematic representation of the number, position, height and staining intensity of bands is presented for the five largest chromosomes of the complement at the four mitotic stages.

Azure Stains↗
PubMed1992

Refractile mycobacteria in Romanowsky-stained bone marrow smears. A comparison of acid-fast-stained tissue sections and Romanowsky-stained smears.

The appearance of mycobacteria was studied in Wright-stained bone marrow preparations of human immunodeficiency virus-infected patients and compared with acid-fast-stained trephine biopsy sections and culture results. Mycobacterium avium complex in Romanowsky-stained preparations may be seen as extracellular and intracellular clear or red refractile beaded rods and nonrefractile "negative images." Refractile mycobacteria were seen in 17 of 20 culture-positive cases. Acid-fast stain of the trephine biopsy demonstrated organisms in only 11 of the 20 cases. Thus, six cases were culture positive and contained refractile rods but had no acid-fast organisms on the trephine biopsy. No false-positive results were seen with Romanowsky stain; the three false-negative results for refractility also were negative with acid-fast stain. Examination of Romanowsky-stained smears or imprints for refractile mycobacteria provides a reliable and sensitive method to identify mycobacteria in this population. Romanowsky-stained bone marrow aspirate and imprint smears should be examined for refractile bacilli when mycobacterial infection is suspected.

Acquired Immunodeficiency Syndrome↗
PubMed2000

Histological identification of Helicobacter pylori: comparison of staining methods.

AIM: To determine whether two recently described staining methods (the modified McMullen's and the Helicobacter pylori silver stain HpSS methods) used for the histological identification of H pylori organisms are superior to two established techniques (the modified Giemsa and anti-H pylori antibody immunostain) in terms of availability, reproducibility, rapidity, sensitivity, and cost. METHODS: Histological sections from 63 paired gastric biopsies from adult patients previously investigated for dyspepsia were stained with the four methods and these were assessed blindly and independently by two observers. Of the 63 patients, 30 were originally negative in all tests for H pylori infection, 30 were positive, and the remaining three cases had discordant results using a combination of five tests (rapid biopsy urease test, urea breath test, culture, serology, and histology). RESULTS: Interobserver agreement was best with the antibody method (98%), followed by the McMullen's (90%), Giemsa (87%), and HpSS (85%). Of the 60 "gold standard" positive and negative cases, 30 were positive by the modified Giemsa stain, 29 by the McMullen's method, 29 by HpSS, and 30 by the antibody stain. However, there were two false positives with the HpSS method. The modified Giemsa is the cheapest and easiest to perform technically. CONCLUSIONS: When H pylori are present, careful examination will almost always reveal them, whichever of these stains is used. However, the modified Giemsa stain is the method of choice because it is sensitive, cheap, easy to perform, and reproducible.

Adult↗
PubMed2001

Atypical Herpes simplex keratitis (HSK) presenting as a perforated corneal ulcer with a large infiltrate in a contact lens wearer: multinucleated giant cells in the Giemsa smear offered a clue to the diagnosis.

PURPOSE: To report a case of atypical herpes simplex keratitis initially diagnosed as bacterial keratitis, in a contact lens wearer. RESULTS: Case report of an 18-year-old woman using contact lenses who presented with pain, redness and gradual decrease in vision in the right eye. Examination revealed a paracentral large stromal infiltrate with a central 2-mm perforation. Corneal and conjunctival scrapings were collected for microbiological investigations. Corneal tissue was obtained following penetrating keratoplasty. Corneal scraping revealed no microorganisms. Giemsa stained smear showed multinucleated giant cells. Conjunctival, corneal scrapings and tissue were positive for herpes simplex virus - 1 (HSV) antigen. Corneal tissue was positive for HSV DNA by PCR. CONCLUSIONS: Atypical HSV keratitis can occur in contact lens wearers. A simple investigation like Giemsa stain may offer a clue to the diagnosis.

Acyclovir↗
PubMed1985

Different banding techniques for the study of bovine chromosomes.

The different banding techniques R-, Q-, C-, NOR- and G-banding allow the identification of individual chromosomes in cattle with greater accuracy than with undifferentiated Giemsa staining. These techniques show characteristic bands on each chromosome and are therefore very useful and reliable for the detection of chromosomal abnormalities.

Animals↗
PubMed1978

A rapid technique for producing silver-stained nucleolus organizer regions and trypsin-giemsa bands on human chromosomes.

A simple and rapid technique is described whereby the nucleolus organizer regions (NORs) of human chromosomes can be differentially stained with silver. This staining is followed by trypsin-Giemsa banding on the same metaphase chromosomes. The metaphases simultaneously exhibit silver-stained NORs and G bands, allowing for the unequivocal identification of all chromosomes and greatly facilitating studies involving the NOR-bearing acrocentrics.

Azure Stains↗
PubMed1986

Detection and quantitation of cellulase by Congo red staining of substrates in a cup-plate diffusion assay.

A simple, inexpensive, and sensitive cellulase assay which allows simultaneous enzyme quantitation for many samples has been developed. Radial diffusion of enzyme into substrate incorporated in an agar gel produces zones of hydrolysis which can be visualized by staining undegraded substrate with Congo red dye. Comparisons were made between this and other commonly used cellulase assays based on accuracy, sensitivity, and ease of use.

Azure Stains↗