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PubMedDate not supplied

Usefulness of estrogen receptor detection using archival Papanicolaou-stained smears.

OBJECTIVE: To examine estrogen receptor (ER) detection using cytologic specimens and to compare the results with those obtained by the dextran-coated charcoal (DCC) method and enzyme immunoassay (EIA). STUDY DESIGN: Immunocytochemical staining was conducted on 60 cases of breast cancer resected at our hospital between April 1993 and November 1997 in which ER had been measured by DCC or EIA. Specimens for immunocytochemical staining were prepared by a cell transfer method using archival Papanicolaou-stained imprint smears, and ER staining was performed by the labeled streptavidin method using an anti-ER monoclonal antibody. These results were compared with those obtained by DCC or EIA. RESULTS: In immunocytochemical staining for ER, positive staining was observed in the nuclei of tumor cells. A good correlation was obtained between the immunocytochemical staining results and biochemical results. Five cases were positive in anti-ER staining but negative in biochemical tests, and two cases were negative in anti-ER staining and positive in biochemical tests. CONCLUSION: Unlike biochemical assays, the immunocytochemical method does not necessitate use of fresh frozen materials and can be performed even using archival Papanicolaou-stained smears. Immunocytochemical study is a highly useful method for routine ER determination.

Adenocarcinoma, Mucinous↗

PubMed1975

Histochemical fiber typing and staining intensity in cat and rat muscles.

In the gastrocnemius muscle of cat and rat, staining for oxidative enzymes differentiated three fiber types (A,B,C) and staining for adenosine triphosphate at pH 9.4 differentiated two fiber types (I, II) with a reliability of 90% and 98%, respectively. In cat 96% and in rat 90% of the fibers were typed identically after staining for nicotinamide adenine dinucleotidelinked lactic dehydrogenase (LDH) and succinic dehydrogenase (SDH). When differentiated by staining for LDH, A and B fibers were of type I. IN RAT, 80-90% OF ALL FIBERS WERE OF TYPE 22, COMPPRISING A, B and C fibers. Type I fibers stained for LDH intensely as did C fibers of type II, but stained intermediately for SDH. The degree of staining was measured by photometry. When fibers were stained for LDH, histograms of density showed three peaks corresponding to A, B and C fibers in cat, but only two peaks corresponding to A and C fibers in rat, In cat and rat, the densities of A, B and C fibers belonged to different populations. In soleus muscle of cat and rat stained for LDH, menadione-linked alpha-glycerophosphate dehydrogenase and adenosine triphosphatase at pH 9.4, the degree of staining differed from thatin any type of fiber in gastrocnemius muscle

Adenosine Triphosphatases↗

PubMed1998

Characteristics of lectin staining patterns assessed by a modified sensitive thermo-method in rat livers with heterologous serum-induced fibrosis.

Lectins are sensitive probes which bind carbohydrate structures specifically. In this study, we modified the lectin staining procedure for sensitive detection of carbohydrate structures in formalin-fixed, paraffin-embedded sections of normal and heterologous serum-induced fibrotic livers. The liver sections were heated in hot distilled water at 100 degrees C for 10 min (thermo-treatment: TT), and then stained with 24 different lectins. In comparison with the results from sections without TT (nonTT), enhanced and/or alternated staining patterns of 19 lectins were demonstrated in sections with TT, and enhanced staining of Vicia villosa agglutinin seen in Kupffer cells was noted. Interestingly, no positive staining was seen with Dolichos biflorus agglutinin, peanut agglutinin or soybean agglutinin (SBA), which recognize O-linked carbohydrate chains, in Kupffer cells of non-TT sections, but strong positive staining was demonstrated in those of TT sections. SBA-positive staining in the cytoplasm of some scattered hepatocytes located in the periportal and perifibrous zones and central zone of pseudolobules was demonstrated only in the fibrotic liver sections with TT. Such findings indicate the heterogeneity of hepatocytes in the liver with fibrosis. Formalin fixation causes masking of lectin binding sites, especially O-linked carbohydrate chains, and TT may recover such masking reactions. TT improved the staining reactions for many lectins in formalin-fixed, paraffin-embedded liver sections, and new staining patterns appear after TT. Modified TT staining procedures may be useful for the diagnosis and prognosis of liver fibrosis.

Animals↗

PubMed1995

A differential staining method for adenohypophyseal cells.

A modification of azan staining (Heidenhain, 1915) for a better differential demonstration than previously of adenohypophyseal cells is reported. Human hypophyses were fixed in Bouin's solution, washed in 70% alcohol, dehydrated and then embedded in paraffin. Sections of about 5 microns thickness were cut and mounted on albumin-coated glass slides. These sections were gently oxidized with a mixture of potassium permanganate and sulphuric acid, and then stained according to the original azan technique; thereafter a procedure of staining with aniline blue was added. With this improvement, human adenohypophyseal cells were clearly classified into six groups on the basis of the color and intensity of staining of the cytoplasm. The alpha-acidophilic cells were stained clear red with azocarmine and the epsilon-acidophilic cells, orange-red with orange G, respectively. With aniline blue, beta-basophilic cells stained a deep blue, while delta-basophilic cells were more weakly stained. The gamma-chromophobic cells were faintly stained red, and the ACTH-cells very faintly blue. Collagenous fibers developed a blue color, while erythrocytes were orange-red to red. This modified method produces much better results in differential staining of the adenohypophyseal cells than either the original azan or Masson-Goldner staining methods.

Adult↗

PubMed1980

Light microscopic and color television image analysis of the development of staining on chlorhexidine-treated surfaces.

Tooth staining with the use of chlorhexidine preparations is the major problem of long term application. Evidence suggests that the staining arises from a cationic/anionic interacation of chlorhexidine with components of certain dietary materials. The purpose of this in vitro study was to compare visually the development of tea and coffee staining on acrylic and tooth specimens treated with chlorhexidine and to follow the development of tea staining on perspex by light microscopy and color television image analysis. All specimens were maintained in their respective beverage for 5 days with test specimens removed three times a day and placed for 2 minutes in an 0.2% chlorhexidine solution. Both test tooth and acrylic specimens showed comparably and markedly increased staining by the beverages compared with control specimens. Color television image analysis of test specimens demonstrated more marked and rapid development of tea staining when studied on a daily basis. Microscopic examination revealed the staining to be made up of small particles of material which increased in size and coalesced with time. Again, marked differences were apparent in the stain on test and control specimens. The results of this in vitro method provided further evidence for a dietary aetiology to chlorhexidine staining and were consistent with clinical findings. Such a method may be useful to assess staining arising from the use of other anti-plaque agents.

Acrylic Resins↗

PubMed1998

Comparison of histologic stains for use in PCR analysis of microdissected, paraffin-embedded tissues.

We evaluated the effect of six different histologic stains on the productivity of PCR amplification of DNA isolated from paraffin-embedded tissue samples. The tissue was collected from glass slides by microdissection techniques, whereby tiny portions of tissue are visually identified through a microscope and selectively resected for subsequent DNA extraction and PCR amplification. We found that the success of PCR amplification depended on the type of histologic stain that was used to facilitate microscopic visualization of the undeparaffinized tissue section. The best results were obtained with methyl green and nuclear fast red, while Wright's stain yielded less PCR product. Two other stains, Evans blue and light-green SF yellowish (also known as the counterstain for geomori methenamine silver stain) yielded sufficient PCR products; however, their staining characteristics did not afford satisfactory visualization of nuclear chromatin to discriminate between benign and malignant cells. Our most significant finding was that a commonly used histologic stain, hematoxylin, failed to produce DNA templates that could be consistently amplified by PCR. In conclusion, it is prudent to avoid hematoxylin stains when preparing tissues as starting material for PCR. Among the remaining five stains that were evaluated, the best choice depends on the differential staining characteristics of the cells to be dissected.

Coloring Agents↗

PubMed1999

Morphologic, cytochemical staining, and ultrastructural characteristics of blood cells from eastern diamondback rattlesnakes (Crotalus adamanteus).

OBJECTIVE: To evaluate light microscopic, cytochemical, and ultrastructural characteristics of blood cells from eastern diamondback rattlesnakes. ANIMALS: 10 healthy snakes. PROCEDURE: Various stains, including Wright-Giemsa, benzidine peroxidase, Sudan black B, chloroacetate esterase, alpha-naphthyl butyrate esterase, acid phosphatase, leukocyte alkaline phosphatase, periodic acid-Schiff with diastase, and toluidine blue, were used to stain leukocytes differentially on multiple blood smears. Electron microscopy also was performed. RESULTS: Lymphocytes were the most commonly observed leukocyte and could be distinguished from thrombocytes, using periodic acid-Schiff stain with diastase. Azurophils also were commonly observed; their granules stained with peroxidase. Eosinophils were not identified; however, 2 morphologic variations of heterophils were seen in the blood of all snakes and were considered the same cell type at different stages of cytoplasmic granule development. Heterophil granules were better preserved, using a one-step Wright-Giemsa method that did not require alcohol fixation prior to staining. Degranulated heterophils were observed in all preparations. CONCLUSIONS: Most leukocytes of eastern diamondback rattlesnakes can be identified easily on Wright-Giemsa-stained preparations. However, hematologic stains that do not require alcohol fixing prior to staining may be preferred for leukocyte evaluation in certain reptiles. A limited degree of heterophil maturation may continue in the blood of healthy snakes. This, along with degranulation of heterophils, may result in a variable staining pattern in this cell type, regardless of the stain used. CLINICAL RELEVANCE: Results provide baseline data for use in hematologic testing in diagnosis of disease and monitoring of treatment of sick or injured snakes.

Animals↗

PubMed2003

[The epidemiology of meconium stained amniotic fluid on hospital basis].

OBJECTIVE: To explore the epidemiology of meconium stained amniotic fluid pero-delivery. METHODS: 6,206 one-fetal delivery cases were divided into meconium stained amniotic fluid and normal amniotic fluid groups. Statistics were analysed on two groups using Cohort study. RESULTS: Incidence of meconium stained amniotic fluid was 16.4%. There was no significant difference on maternal medical complication between two groups. The percentage of parity, gestational week > or = 42 weeks and big birth weight were higher in meconium stained amniotic fluid group than that in normal amniotic fluid group (P < 0.001). The average Apgar score in meconium stained amniotic fluid group was lower than that in normal amniotic fluid group (P = 0.001). The percentage of stillbirth, low birth weight and transferred newborn care unit in meconium stained amniotic fluid group were higher than that in normal amniotic fluid group (P < 0.001). CONCLUSIONS: The relative factors on meconium stained amniotic fluid were maternal parity, gestational weeks > or = 42 weeks and big birth weight. No correlation between meconium stained amniotic fluid and maternal medical complication. Morbidity of newborn baby was higher in meconium stained amniotic fluid. Meconium stained amniotic fluid is a important clinical factor on evaluating pregnancy outcome.

Adult↗

PubMed2004

Comparative tooth whitening and extrinsic tooth stain removal efficacy of two tooth whitening dentifrices: six-week clinical trial.

OBJECTIVE: The objective of this six-week, examiner-blind clinical study was to assess the tooth whitening and extrinsic tooth stain removal efficacy of a new dentifrice delivering 1.0% hydrogen peroxide, 0.243% sodium fluoride, and sodium tripolyphosphate in a high-cleaning silica base (Test Dentifrice), relative to that of commercially available hexametaphosphate-containing whitening dentifrice (Positive Control Dentifrice). METHODOLOGY: Following a baseline oral soft tissue examination and scoring of extrinsic tooth stain and tooth shade, qualifying adult male and female subjects from the Harrisburg, Pennsylvania area were randomized into either the Test or Positive Control Dentifrice group. The two groups were balanced for gender, extrinsic tooth stain, and tooth shade scores. All subjects were provided their assigned dentifrice and a soft-bristled adult toothbrush for home use. Subjects were instructed to brush their teeth for two minutes twice daily (morning and evening) using only the dentifrice provided, and to refrain from using any other oral hygiene products for the entire six weeks of the study. There were no restrictions regarding diet or smoking habits during the course of the study. Oral soft tissue, extrinsic tooth stain, and tooth shade assessments for each subject were repeated after two and six weeks of product use. All statistical tests were two sided and employed a level of significance of alpha = 0.05. RESULTS: Fifty-six (56) subjects complied with the protocol and completed the entire study. Compared to baseline at both the two- and six-week examinations, the Test Dentifrice group had statistically significant reductions in extrinsic tooth stain area and intensity, and statistically significant mean shade rank reductions, with a six-week reduction of 4.81. In contrast, at six weeks, the Positive Control dentifrice had a statistically significant increase in tooth stain area, a non-significant increase in tooth stain intensity, and a statistically significant mean 1.40 shade rank score reduction. At both the two- and six-week examinations, the Test Dentifrice group had statistically significantly lighter tooth shades and statistically significantly lower levels of extrinsic tooth stain area and intensity than the Positive Control Dentifrice group. CONCLUSION: A new whitening dentifrice delivering 1.0% hydrogen peroxide, 0.243% sodium fluoride, and sodium tripolyphosphate in a high-cleaning silica base has significant efficacy in tooth whitening and extrinsic stain removal, and provides a statistically significantly greater level of efficacy for both tooth whitening and extrinsic tooth stain removal than the commercially available Positive Control Dentifrice.

Adolescent↗

PubMed2004

Efficacy of a modified ultrafast Papanicolaou (UFP) stain for breast aspirates.

Rapid assessment of fine needle aspiration smears has become popular. The goal is minimal time required and fastest turn around time for test results in breast aspirates. In the present study, fine needle aspiration smears from 100 cases of breast lump were stained by modified ultrafast Papanicolaou stain which consisted of the following changes from ultrafast Papanicolaou stain: (i) Use of Gill 's haematoxylin instead of Richard Allan haematoxylin, (ii) omission of orange G from cytostain. Wet fixed smears subjeted to May-Gruenwald and Giemsa stain and an air dried smear subjected to modified ultrafast Papanicolaou stain were compared. Smears stained by modified stain showed transparent cells with crisp nuclear features in a blood free background. Total time required in this staining was 130 seconds. Out of 100, diagnosis was inadequate in 5 cases. In 95 cases cytological diagnosis was as follows: 60 benign, 27 ductal malignancy and 8 of inflammatory lesions. There was 100% agreement in cytologic diagnosis offered on smears stained by May-Gruenwald and Giemsa and modified ultrafast Papanicolaou stain.

Biopsy, Fine-Needle↗

PubMed2005

Efficacy of a novel brush head in the comparison of two power toothbrushes on removal of plaque and naturally occurring extrinsic stain.

OBJECTIVES: To compare the safety and efficacy of an oscillating/pulsating power toothbrush (Oral-B ProfessionalCareTM 7000; PC 7000) fitted with either the standard FlexiSoft (PC 7000/EB17) brush head or the novel Pro Polisher (PC 7000/ EB-Prophy) and a high-frequency toothbrush (Philips Sonicare Elite; SE), in their relative ability to remove plaque and naturally occurring extrinsic dental stain over a six-week period. METHODS: This randomised, examiner-blind, parallel group study involved 90 healthy subjects from a general population. All subjects received a baseline plaque (Turesky et al. modified Quigley-Hein Plaque Index), stain (Lobene Stain Index) and tooth shade (VITAPAN Shade Guide) [Vita] assessment and an oral tissue examination. After training in the use of their randomly assigned device, subjects were instructed to brush twice daily for 2 min and returned after 3, 4 and 6 weeks of product use for a repeat of each clinical assessment. RESULTS: Reductions from baseline in mean plaque and extrinsic dental stain scores were significant at Weeks 3, 4 and 6 in all three treatment groups. By Week 6, mean reductions from baseline in whole mouth plaque scores were 32%, 27% and 14% in the PC 7000/EB-Prophy, PC 7000/EB17 and SE groups, respectivety. For the body of the tooth, mean reductions from baseline at Week 6 in total stain were 89%, 89% and 80%, respectively. Between treatment group comparisons consistently revealed that the PC 7000 toothbrush plus the EB-Prophy or EB17 brush head removed significantly more plaque and extrinsic stain (total stain, stain area and stain intensity) than the SE toothbrush at 3, 4 and 6 weeks. The EB-Prophy group had a greater proportion of subjects showing a 2-3+ change in Vita shade scores at each time point compared to the other two brushes; at Week 6 the proportions were 67% in the PC 7000/EB-Prophy group, 30% in the PC 7000/EB17 group, and 7% in the SE group. The PC 7000/EB17, PC 7000/EB-Prophy and the SE were found to be safe as used in the study. CONCLUSIONS: The oscillating/rotating/pulsating PC 7000 (fitted with either the standard EB17 or novel EB-Prophy brush head) is more effective at removal of plaque and naturally occurring extrinsic tooth stain, and the PC 7000 plus EB-Prophy in the improvement of tooth shade, than the high-frequency SE toothbrush.

Adolescent↗

PubMed2005

Efficacy of a novel brush head in the comparison of two power toothbrushes on removal of plaque and naturally occurring extrinsic stain.

OBJECTIVES.: To compare the safety and efficacy of an oscillating/pulsating power toothbrush (Oral-B ProfessionalCaretrade mark 7000; PC 7000) fitted with either the standard FlexiSoft (PC 7000/EB17) brush head or the novel Pro Polisher (PC 7000/EB-Prophy) and a high-frequency toothbrush (Philips Sonicare(R) Elite(R); SE), in their relative ability to remove plaque and naturally occurring extrinsic dental stain over a six-week period. METHODS.: This randomised, examiner-blind, parallel group study involved 90 healthy subjects from a general population. All subjects received a baseline plaque (Turesky et al. modified Quigley-Hein Plaque Index), stain (Lobene Stain Index) and tooth shade (VITAPAN(R) Shade Guide) [Vita] assessment and an oral tissue examination. After training in the use of their randomly assigned device, subjects were instructed to brush twice daily for 2 min and returned after 3, 4 and 6 weeks of product use for a repeat of each clinical assessment. RESULTS.: Reductions from baseline in mean plaque and extrinsic dental stain scores were significant at Weeks 3, 4 and 6 in all three treatment groups. By Week 6, mean reductions from baseline in whole mouth plaque scores were 32%, 27% and 14% in the PC 7000/EB-Prophy, PC 7000/EB17 and SE groups, respectively. For the body of the tooth, mean reductions from baseline at Week 6 in total stain were 89%, 89% and 80%, respectively. Between treatment group comparisons consistently revealed that the PC 7000 toothbrush plus the EB-Prophy or EB17 brush head removed significantly more plaque and extrinsic stain (total stain, stain area and stain intensity) than the SE toothbrush at 3, 4 and 6 weeks. The EB-Prophy group had a greater proportion of subjects showing a 2-3+ change in Vita shade scores at each time point compared to the other two brushes; at Week 6 the proportions were 67% in the PC 7000/EB-Prophy group, 30% in the PC 7000/EB17 group, and 7% in the SE group. The PC 7000/EB17, PC 7000/EB-Prophy and the SE were found to be safe as used in the study. CONCLUSIONS.: The oscillating/rotating/pulsating PC 7000 (fitted with either the standard EB17 or novel EB-Prophy brush head) is more effective at removal of plaque and naturally occurring extrinsic tooth stain, and the PC 7000 plus EB-Prophy in the improvement of tooth shade, than the high-frequency SE toothbrush.

Journal Article↗

PubMed1994

To approve or not to approve? That is the question. Ethical concerns over the regulation of immunohistochemical stains.

Over the past two decades, immunohistochemical stains have evolved to become an essential part of the practice of surgical pathology. An extensive literature documents significant diagnostic and prognostic applications, and the pathology journals add new applications and new tests almost on a daily basis. The antibodies that provide the basis of immunohistochemical stains fall under the purview of the Food and Drug Administration (FDA). The vast majority of these antibodies have not been submitted for FDA approval or clearance, but nonetheless are widely employed in practice. The FDA, therefore, faces a major dilemma in attempting to fulfill its legislative mandate. Lack of adequate regulation may permit the use of unproven or poor quality reagents. On the other hand, overzealous regulation will preclude the use of large numbers of stains that are now integral to the diagnostic process. If ethical considerations truly are governed by following the course that leads to the greatest good for the greatest number, then the evidence suggests that overzealous regulation should be avoided. The argument is presented that immunohistochemical stains should be treated no differently from other special stains already in use in surgical pathology. With all tissue stains, including immunohistochemical stains, the pathologist makes the diagnosis by interpreting morphologic features and staining patterns: the stain alone is of no value to clinician or patient, rather it is the pathologist's report that establishes the diagnosis. It is suggested that a certification process that ensures quality and technical validity, somewhat analogous to that already in existence for biological stains, would be sufficient to meet the needs of the pathologist, providing ongoing benefits to patient care, with minimal risk.

Drug and Narcotic Control↗

PubMed1997

Quantification of immunohistochemical staining by color translation and automated thresholding.

OBJECTIVE: To develop a method of quantitating immunohistochemical staining using diaminobenzidine (DAB) and hematoxylin by means of color image analysis. STUDY DESIGN: Red-green-blue (RGB)-filtered grayscale values from images from microscopic slides of mouse jejunum, stained with DAB, hematoxylin or both were analyzed using the public domain program NIH Image. Based on the correlations between the R-G- and B-filtered grayscale values, a simple translation algorithm using the RGB information was developed, providing the option for separation of DAB only- and double-stained areas from hematoxylin only-stained areas by means of automated thresholding. The method was tested by staining mouse jejunum for the growth factors EGF, TGF-alpha and TGF-beta 1-3 using immunohistochemical techniques. RESULTS: A good separation of DAB- and double-stained pixels from hematoxylin-stained pixels was achieved, with misclassification of only 2.4% of the pixels as compared to 34% misclassification using automated thresholding of the blue component of the RGB image, the untransformed grayscale images with the most contrast for DAB- and non-DAB-stained areas. Significant differences in relative areas stained and mean specific optical density for the growth factors in mouse jejunal crypts and villi were observed. CONCLUSION: The image analysis method described offers the possibility of objective determination of stained area in histologic slides with the commonly used DAB and hematoxylin chromophores. It shows that reproducible and objective measurements can be made based on RGB true color images acquired using a simple microscope and video camera setup and the public domain program NIH Image.

3,3'-Diaminobenzidine↗

PubMed1997

The usefulness of the reticulin stain in the differential diagnosis of liver nodules on fine-needle aspiration biopsy cell block preparations.

We reviewed fine-needle aspiration biopsy (FNAB) cell blocks of hepatocellular carcinoma (HCC) (n = 16) and benign hepatic processes (n = 16) to evaluate the significance of reticulin staining (Gomori stain) in combination with standard cytomorphologic and architectural criteria. We analyzed the staining pattern using semiquantitative grading: normal, variable, decreased, or virtually absent. Also, we graded the cell thickness of the hepatic trabeculae as greater than or less than three cells. Fourteen of 16 biopsy specimens of benign processes demonstrated a normal reticulin framework, with staining outlining hepatic trabeculae less than three cell layers in thickness. Staining was markedly decreased in one case of steatosis and virtually absent in one case of cirrhosis. In contrast, all of the 16 HCCs demonstrated either a virtually absent (7 of 16), decreased (6 of 16), or variable (3 of 16) reticulin staining pattern, with thickened trabeculae greater than three cell layers. We conclude that the reticulin stain is a useful adjunct in the differential diagnosis of liver nodules on FNAB cell block preparations and that it is particularly useful in distinguishing HCC from benign hepatic processes. Virtually absent or decreased reticulin staining and staining outlining trabeculae greater than three cells in thickness support the diagnosis of HCC. Normal reticulin staining outlining well-defined hepatic trabeculae less than three cell layers in thickness supports the diagnosis of a benign hepatic process.

Biopsy, Needle↗

PubMed1998

Fontana-Masson--stained tissue from culture-proven mycoses.

BACKGROUND: The Fontana-Masson stain can be used to detect melanin of dematiaceous fungi such as Curvularia lunata and Bipolaris hawaiiensis in tissue. This stain is often used to identify fungi that may not have an evident brown color in hematoxylin-eosin-stained sections. However, it is not clear whether hyphae of nondematiaceous fungi can be stained with the Fontana-Masson procedure. METHODS: Paraffin sections from culture-proven mycoses caused by species of Aspergillus, Candida, Fusarium, Pseudallescheria, Trichosporon, zygomycetes, and dematiaceous fungi were stained by the Fontana-Masson procedure. Extent of stain intensity and its distribution in fungal elements in tissue were evaluated by means of an intensity scale based on Cryptococcus neoformans as a positive control. RESULTS: Many Aspergillus spp, some zygomycetes, and all of the dematiaceous fungi showed strong intensity, whereas other fungi showed either weak intensity or no staining. CONCLUSION: Fungal hyphae other than dematiaceous fungi occasionally can be stained with the Fontana-Masson procedure. The morphology of positively stained hyphae must be carefully evaluated, especially when they are hyaline in hematoxylin-eosin-stained tissue sections.

Aspergillus↗

PubMedDate not supplied

Microwave-assisted staining procedures in routine histopathology.

The use and practicability of microwave-assisted staining procedures in routine histopathology over more than three years has been evaluated. A domestic microwave oven was used to speed up the following staining procedures: Haematoxylin-Eosin (for frozen sections), Romanowsky-Giemsa, Periodic acid-Schiff (PAS), Ziehl-Neelson, Papanicolaou, Feulgen and Grocott--stain on buffered formalin fixed sections or cytologic smears. These staining procedures can be made highly reproducible providing; (1) Staining vessels are placed in the same position inside the oven; (2) Accurate timing in seconds is observed. Microwave-assisted staining procedures are equal to or even superior to those of the standard methods. Staining times can be reduced to 2%-10% of the conventional staining procedures. The basic staining protocols are presented.

Azure Stains↗

PubMed1982

Spreading and staining of human metaphase chromosomes on aminoalkylsilane-treated glass slides.

The properties of aminoalkylsilane-treated glass slides for the preparation of metaphase spreads and their staining quality have been studied and compared with those of slides which had only been cleaned in ethanol/ether. The parameters investigated were: (1) the average area of metaphases from cultures of blood from both healthy donors and haematology patients; (2) the influence of the positively charged 'coating' on the quality of quinacrine- and Giemsa-banding patterns; (3) non-specific background staining for these banding methods; (4) the number of metaphases as compared to the number of interphase cell nuclei per area of preparation; and (5) the Feulgen-staining intensities of chromosomes and chicken erythrocyte nuclei. The quality of metaphase preparations and the differential staining of chromosomes is better on aminoalkysilane-treated glass slides than that of preparations on routinely cleaned normal microscope slides. In the preparations on aminoalkylsilane-treated slides, the distribution of the cells over the glass surface is more homogeneous; and no influence could be detected on the relative frequency of metaphases as compared to the number of non-divided cell nuclei; the average area per metaphase is increased by about 10% and consequently the number of overlapping chromosomes is decreased. Preparations on aminoalkylsilane-treated glass, after Q-, G- and DAPI-banding procedures, always showed less binding of the staining compounds to the glass slide (a cleaner background) than those on routinely cleaned microscope glass slides. The Feulgen-pararosaniline staining intensities of human metaphase chromosomes and chicken erythrocyte nuclei are the same on aminoalkylsilane-treated slides and on routinely cleaned glass slides. Furthermore, the reproducibility and constancy of quinacrine banding was improved by development of an equilibrium staining method which does not require a washing procedure. The medium, containing 0.002% quinacrine, allows optimal staining results to be obtained for microphotography purposes within 30 min of staining (for visual inspection at least 90 min is required) and is used as the embedding medium. In combination with aminoalkylsilane-treated glass slides, this procedure leads to a clean background and reproducible banding patterns of excellent quality, the results being better and more constant than those of methods described before.

Azure Stains↗