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PubMed1987

Linear immunofluorescence staining of the cutaneous basement membrane zone produced by pemphigoid antibodies: the result of hemidesmosome staining.

Bullous pemphigoid autoantibodies bind the basement membrane zone of stratified squamous epithelium in a linear pattern, as shown by indirect immunofluorescence; however, other patterns of staining, such as tubular (with convolutions), cytoplasmic, and membranous, have been noted. Recently, by using indirect immunoelectron microscopy, we have shown that bullous pemphigoid antibodies bind specifically to hemidesmosomes. The purpose of this investigation was to correlate the various patterns of basement membrane zone staining by bullous pemphigoid antibodies by immunofluorescence and to correlate these patterns with the ultrastructural findings. We employed adult human skin, neonatal human foreskin, neonatal mouse skin, and rabbit cornea as substrates for electron microscopy and indirect immunofluorescence using bullous pemphigoid serum. For indirect immunofluorescence, cryosections were obtained at vertical, oblique, and horizontal planes with respect to the basement membrane zone. We show that the immunofluorescence band of basement membrane zone staining results from the coalescence of fluorescence from individual hemidesmosomes. We also show that the pattern of basement membrane zone staining depends on the ultrastructural morphology of the basement membrane zone in each tissue and on the angle of sectioning.

Animals↗
PubMedDate not supplied

Protein staining influences the quality of mass spectra obtained by peptide mass fingerprinting after separation on 2-d gels. A comparison of staining with coomassie brilliant blue and sypro ruby.

When separating protein mixtures on 2-D gels for proteomics purposes, fluorescent staining is superior in sensitivity and linear response as compared to Coomassie Brilliant Blue (CBB) and silver staining, respectively. We have compared the quality of mass spectra for proteins obtained from gels stained with CBB and SYPRO Ruby (SR) and found significant differences. These differences can be seen both in inferior signal/noise ratios and number of peptides detected with the fluorescent stain.

Animals↗
PubMed1991

Shift in the megakaryocyte ploidy in MDS patients: microcytofluorometry with DAPI staining after destaining of Wright-Giemsa stain.

We applied DAPI (4',6-diamidino-2-phenylindole) staining to the determination of nuclear DNA content in single megakaryocytes in 12 normal subjects and 12 patients with myelodysplastic syndrome (MDS). After the megakaryocytes had been identified on Wright-Giemsa stained smear and classified according to modified Feinendegen's classification, they were photographed. Then Wright-Giemsa stain was removed by immersion in 50% ethanol at 37 degrees C for 1 h and 100% methanol at 37 degrees C for 1 h. The specimens were then stained with DAPI solution (DAPI 0.01 mg/ml, pH 7.4 Tris-EDTA-2Na buffer solution and 0.01 M 2-mercaptoethylamine hydrochloride mixed at the ratio of 0.5:98.5:1.0) for more than 30 min. The amount of nuclear DNA in the previously identified megakaryocytes was measured by microcytofluorometry. The maximum population of megakaryocytes ploidy was in 16N in normal subjects, 8N in 10/12 MDS patients, and 4N in the remaining two patients. These findings suggest impairment of the development of the megakaryocytes nucleus in the MDS patients.

Adolescent↗
PubMed1976

The influence of the embedding medium when staining for electron microscopy: the penetration of stains into plastic sections.

Using a sectioned-section procedure it was found that only a few structures, e.g. the connective tissue elements, were deeply penetrated by phosphotungstic acid (PTA) and uranyl acetate (UA). Reynold's lead citrate appeared to penetrate more generally. Using stains selective for the embedding medium, and also observing the surfaces of shadowed sections, it was concluded that structures readily penetrated by polar stains were only poorly infiltrated by non-polar embedding media. It is argued that this differential infiltration leading to differential penetration by polar stains largely controls the pattern of staining of ultrathin sections by PTA and UA.

Animals↗
PubMed1990

Comparative study of immunocytochemical staining versus Giemsa stain for detecting dysmegakaryopoiesis in myelodysplastic syndromes (MDS)

23 patients with myelodysplastic syndromes (MDS) and 8 normal controls were analyzed for dysmegakaryopoiesis (DMP) in the bone marrow by alkaline phosphatase anti-alkaline phosphatase (APAAP) technique and by conventional May-Giemsa staining. In the immunocytochemical study, monoclonal antibody (MoAb) against glycoprotein (GP) IIb/IIIa was utilized to demonstrate megakaryocytic cells. 91% (21/23) of MDS cases were detected as having DMP by APAAP method, while only 52% (12/23) were detectable by Giemsa stain. There were difficulties in recognizing small micromegakaryocytes (micro MKs), designated as type 1 atypical MKs, by Giemsa staining. Furthermore, megakaryoblasts (MKBs) were detectable only by APAAP technique. In 8 normal controls, no type 1 and type 3 atypical MKs (round shaped multinuclear MKs) were observed either by Giemsa staining or by the APAAP method, suggesting that they are a distinctive feature of MDS. These results indicate the necessity of immunocytochemical technique for accurate recognition of DMP in MDS.

Anemia↗
PubMed1983

Comparative study of subculture, Gram staining and acridine orange staining for early detection of positive blood cultures.

In view of the importance of a rapid aetiological diagnosis in septicaemia, we compared the results of subculture, Gram staining and acridine orange staining in the detection of positive blood cultures. The study was based on 1013 blood cultures of which 138 were positive by culture. The three techniques were applied 12 h after the specimen was taken in 210 instances, at 24 h in 540 instances and after 48 h in 525. We were able to demonstrate the value of direct examination. Staining with acridine orange yields more positive results than Gram staining and is also simpler.

Acridine Orange↗
PubMed1977

Differential staining of calcified tissues in plastic embedded microtome sections by a modification of Movat's pentachrome stain.

Movat's pentachrome I stain has been adapted and modified as a stain for undecalcified bone sections. After embedding in methyl methacrylate, this procedure yields consistently good results, with an excellent and colorful contrast between mineralized and unmineralized compartments of both cartilage and bone. In addition, osteoblasts, osteoclasts, and other cells and tissue components can easily be differentiated. The staining properties of the lacunar wall surrounding the osteocytes are considered to reflect various states of osteocytic activity. The method is especially useful for the study of bone growth and bone repair, and as a stain for conventional histomorphometry and computer-assisted image analysis in bone biopsies.

Animals↗
PubMed2000

Quantitative chromatics analysis for computer imaging of cytologic subtypes of lung cancer stained by Papanicolaou stain.

OBJECTIVE: To determine the role of quantitative chromatics analysis in the classification of subtypes of lung cancer stained by Papanicolaou stain. STUDY DESIGN: By means of computer image analysis, 60 keratinized squamous carcinoma cells (KSCC), 88 nonkeratinized squamous carcinoma cells (NKSCC) and 150 adenocarcinoma cells (ACC) from lung cancer in sputum smears stained by Papanicolaou stain were analyzed and distinguished based on quantitative colorimetry. The features measured were the content of three primary colors, red (R), green (G) and blue (B) and the coefficients of R, G and B (r, g and b, respectively). Hue, saturation, brightness and gray level were also measured. A stepwise discriminant analysis was carried out. RESULTS: The values of R, G and B and r, g and b, hue and saturation in NKSCC and ACC were significantly different from those of KSCC, and the changes in the three primary colors were more sensitive than those in the gray level. Computer assessment based on three primary color coefficients, hue and saturation yielded accuracy of distinguishing KSCC from NKSCC and KSCC from ACC of 95.2% and 95%, respectively. CONCLUSION: Quantitative analyses of R, G and B and r, g, b and hue and saturation are valuable in distinguishing KSCC from NKSCC and ACC.

Adenocarcinoma↗
PubMed2001

[A study on pathological changes of closed diffuse brain injury in rats with H.E., esterification-silver stain and albumin immunohistochemical stain].

OBJECTIVE: To observed the pathological changes of closed diffuse brain injury in the rats died immediately and 15 min to 5 days after the injury. METHODS: H.E. staind and esterification-silver stain were applied to investigate the closed diffuse brain injury. RESULTS: In rats died immediately after the concussive injury, a number of shrunken neurons(type I change), distended neurons(type II change) and wave-like nerve fibers were identified in the brain tissue, especially in brain-stem. At 2 h and 8 h after injury, brain edema and axonal swelling appeared clearly in the cortex and white matter, especially in brain-stem. At post-traumatic 8 h and 24 h, the axonal retraction balls began to appear. The amount of neurons undergoing type I and II changes and constraction balls increased along with the survivor time. After 4 days and 5 days, brain edema alleviated, but the retraction balls and axonal swelling still existed. With Esterifica-tion silver stain, the above changes of neurons and nerve fibers were more obvious. With ABC stain, the distribution of albumin(Al) was extended from the perio-vascular area to diffuse distribution. Al positive staining were more obvious in injuried neurons and nerve fibers. CONCLUSION: The distribution of the concussive damage in the brain are coup, contra-coup and centripental.

Albumins↗
PubMed1992

Does microwave irradiation have other than thermal effects on histological staining of the mammalian CNS? A light microscopical study of microwave stimulated staining under isothermal conditions in man and rat.

The question whether or not microwave irradiation exerts other than thermal effects on histological staining is still a matter of controversy. The present study was undertaken to reveal or reject such a so far hypothetical non-thermal irradiation effect. A device was developed, which enables exposure of histological sections or tissue pieces to microwave irradiation under isothermal conditions, i.e. with synchronous removal of the internal heat produced. Three classical neuroanatomical staining methods were tested on human and rat CNS. As control, identical procedures were performed without simultaneous microwave irradiation. The experiments were performed at three different temperature levels ranging from 5 to 50 degrees C. In none of the cases studied was a light microscopically appreciable difference observed between the microwave and non-microwave versions of a stain at the same temperature. The hypothesis of a separate non-thermal effect of microwave irradiation on histological staining is therefore rejected.

Animals↗
PubMed2004

Evaluation of applying feulgen stain for DNA analysis on destained hematoxylin-eosin-stained cytologic smears.

OBJECTIVE: To evaluate the feasibility and reliability of DNA analysis performed on the original hematoxylin-eosin (HE)-stained cytologic specimens by destaining the slides and restaining with the Feulgen method. STUDY DESIGN: Image cytometric analysis of DNA ploidy status was performed in a comparative study on 20 cytologic preparations from 10 nasopharyngeal carcinomas. Ten smears were stained directly by the Feulgen method, and the others were Feulgen stained after HE destaining. RESULTS: There was 90% overall concordance in DNA determination and a good correlation (r = .97, P < .001) between the DNA indices determined by the 2 methods. Discordance was probably due to tumor heterogeneity. CONCLUSION: This study demonstrated that image cytometric DNA analysis on previously routinely HE-stained cytologic preparations is feasible and reliable. This method permits retrospective studies on archival cytologic material from patients with long term follow-up data.

Adult↗
PubMed2006

Correlation between the papanicolaou stain and the Wright-Giemsa stain in body fluids: a quality assurance study.

OBJECTIVE: To compare the use of Papanicolaou and Wright-Giemsa stains for the evaluation of body fluids in cytology and hematology laboratories and determine whether other factors account for discrepancies in diagnosis. STUDY DESIGN: We retrospectively reviewed cytopathology reports of peritoneal, pleural, and cerebrospinal fluids received by hematology and cytology laboratories for 1 year. Cases were divided into 3 categories-benign, atypical, and malignant--and slides of discrepant diagnoses were reviewed. RESULTS: During this period, 198 of 3212 (0.61%) cases received by the hematology laboratory and 252 of 4402 (0.57%) cases received by the cytology laboratory were diagnosed as malignant or atypical. Of 3212 cases simultaneously received by the cytology and hematology laboratories, 17 diagnosed as malignant by hematology were diagnosed benign by cytology (sensitivity 96%). Sixteen cases diagnosed as malignant by cytology were diagnosed as benign by hematology (sensitivity 97%). No benign cases were diagnosed as malignant (specificity 100%). Review of the glass slides of the discrepant cases revealed 8 cases undercalled by hematology and 7 cases undercalled by cytology. CONCLUSION: Papanicolaou stain is superior for carcinoma and Wright-Giemsa stain for hematopoietic disorders, but used together they may reduce false negative results. Delays in processing, staining technique, and interobserver variability contribute to discrepancy.

Body Fluids↗
PubMed1986

The staining properties of halo- and alkyl-thiazolylazophenol derivatives in histochemical staining of cadmium.

In the continuous research for a more sensitive chelator for cadmium, halothiazolylazodiethyaminophenol and alkylthiazolylazophenol derivatives were synthesized, and their staining sensitivity for cadmium chelates were examined with respect to a substituent group. 2-(4-Methyl- and 4,5-dimethyl-2-thiazolylazo)-5-diethylami-nonphenol showed low staining density in the 005-sections of cadmium, to date they showed the highest staining sensitivity for cadmium among the thiazolylazophenol analogs. The appropriate combination of substituent groups as based on their electron donative strength may be the decisive factor in determining the staining sensitivity of thiazolylazophenol analogs.

Animals↗
PubMed1987

Estrogen staining in breast carcinoma by PAP methods compared to CEA and ferritin staining.

The aims of this paper are to demonstrate the stainability of estrogen, CEA, and ferritin in breast carcinomas, fibroadenomas, and fibrocystic diseases; to examine whether the findings of endogenous estrogen using the immunohistochemical detection method are related to estrogen receptor (ER) assays; and to determine whether the stainability of estrogen, CEA, and ferritin were related to the prognosis of breast carcinomas. In breast cancer, the stainability of estrogen using the peroxidase-antiperoxidase (PAP) method was positively correlated with the dextran-coated charcoal (DCC) assay for ER. In breast cancers, the percentage of positive staining was 46% for estrogen, 48% for CEA, and 47% for ferritin. With all three stains, significant differences were observed between cancer and benign diseases. Cases that were both positive for estrogen staining and negative for CEA showed a good prognosis after the recurrence of disease. Our data suggest that the immunohistochemical staining of estrogen, CEA, and ferritin might predict the biological behavior of breast carcinomas and be a prognostically useful indicator of breast cancer patients.

Adenofibroma↗
PubMed1989

Differential staining of tick chromosomes: techniques for C-banding and silver-staining and karyology of Rhipicephalus sanguineus (Latreille).

A new karyotypic procedure is described for tick chromosomes along with modifications of accepted techniques for C-banding and silver-staining. The staining procedures produce patterns with staining limited to the centromeric regions and the nucleolar organizer regions (NORs). The possible reasons for the concordant patterns are discussed based on the staining characteristics of the 2 techniques. The karyotype of Rhipicephalus sanguineus possesses 20 acrocentric autosomes with a single pair of NORs located proximal to the centromere on the largest autosomal pair. The sex-determining system is XX:XO with the X chromosome being the largest chromosome in the complement and subtelocentric.

Animals↗
PubMedDate not supplied

Quantitative computerized analysis of silver-stained and Coomassie Blue-stained two-dimensional protein maps.

Using the two-dimensional gel electrophoresis and two systems for computer analysis of the resulting maps, we obtained a good quantitative reproducibility of individual spots of silver-stained and Coomassie Blue-stained gels from mouse fetus liver samples. The GelScan-XL system was established for the evaluation of silver-stained gels. The Gel-Image software has shown to be capable of the analysis of Coomassie Blue-stained gels and autoradiographs. For these systems the linear relationship between defined concentrations of protein loaded on the gel and the resulting integrated intensities of spots were examined. The results show that both systems are efficient to detect quantitative changes in protein expression correlated with malformations in mouse fetuses.

Animals↗