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PubMed1996

Detection of myocardial infarction by immunohistological staining for C9 on formalin fixed, paraffin wax embedded sections.

AIMS: To evaluate an immunohistological stain for complement component C9 as a method of detecting early myocardial infarction and to compare this with (1) an enzyme histochemical method and (2) conventional histological staining. METHODS: (1) Eight hearts taken at necropsy were stained using the nitroblue tetrazolium/phenazine methosulphate method and an immunohistological stain for C9. (2) Twenty five hearts from cases of suspected or confirmed myocardial infarction and 25 from cases without conventional evidence of infarction were stained for C9 and by haematoxylin and eosin. RESULTS: (1) The histochemical method indicated myocardial necrosis in five hearts and the C9 method in seven, all of which had clinical evidence of myocardial damage or a reason for it. The histochemical method required fresh myocardium, was difficult to use and was difficult to interpret. (2) Of 25 hearts with suspected or confirmed infarction, 24 were stained by the C9 method. Staining with haematoxylin and eosin showed infarction in 16 of these, all with infarcts at least 24 hours old; the other eight had clinical evidence of infarction less than 24 hours old. The heart not stained by C9 was from a patient who, on review, had no evidence of infarction. Of the 25 control hearts, none had infarction on staining with haematoxylin and eosin, but three were stained by the C9 method. These three were from patients with septicaemia or another reason for myocardial damage. CONCLUSIONS: The immunohistological method for C9 is a simple, reliable and sensitive method for the detection of early myocardial necrosis that could be used on formalin fixed, paraffin wax embedded necropsy material. This had advantages over a histochemical method and conventional staining with haematoxylin and eosin.

Adult↗

PubMed2005

Analyzing patterns of staining in immunohistochemical studies: application to a study of prostate cancer recurrence.

BACKGROUND: Immunohistochemical studies use antibodies to stain tissues with the goal of quantifying protein expression. However, protein expression is often heterogeneous resulting in variable degrees and patterns of staining. This problem is particularly acute in prostate cancer, where tumors are infiltrative and heterogeneous in nature. In this article, we introduce analytic approaches that explicitly consider both the frequency and intensity of tissue staining. METHODS: Compositional data analysis is a technique used to analyze vectors of unit-sum proportions, such as those obtained from soil sample studies or species abundance surveys. We summarized specimen staining patterns by the proportion of cells staining at mild, moderate, and intense levels and used compositional data analysis to summarize and compare the resulting staining profiles. RESULTS: In a study of Syndecan-1 staining patterns among 44 localized prostate cancer cases with Gleason score 7 disease, compositional data analysis did not detect a statistically significant difference between the staining patterns in recurrent (n = 22) versus nonrecurrent (n = 22) patients. Results indicated only modest increases in the proportion of cells staining at a moderate intensity in the recurrent group. In contrast, an analysis that compared quantitative scores across groups indicated a (borderline) significant increase in staining in the recurrent group (P = 0.05, t test). CONCLUSIONS: Compositional data analysis offers a novel analytic approach for immunohistochemical studies, providing greater insight into differences in staining patterns between groups, but possibly lower statistical power than existing, score-based methods. When appropriate, we recommend conducting a compositional data analysis in addition to a standard score-based analysis.

Biomarkers, Tumor↗

PubMedDate not supplied

Relative value and cost-effectiveness of culture and special stains in fine needle aspirates of the lung.

OBJECTIVE: To define the role of microbiologic stains and culture in lung fine needle aspiration (FNA) specimens. STUDY DESIGN: All lung FNA specimens over a nine-year period, with results of both culture and microbiologic stains (Gram's, methenamine silver and acid fast) were reviewed and correlated with clinical information. RESULTS: Sixty-five cases were identified; 13 cases represented clinically significant infections (20%). Gram's stain identified 3 infections and had 1 false positive result, while culture identified 7 infections and had 9 false positive results. However, all false positive cultures represented easily identifiable contaminants, and eight of nine cases had no associated acute inflammation or necrosis. Aspergillus species were detected in four cases by Papanicolaou and silver stain, while culture was positive in only one case. Coccidioides immitis was detected by both Papanicolaou stain and culture in one case. A single case of Mycobacterium tuberculosis was identified by both culture and acid-fast stain. While culture appeared more cost-effective than Gram's stain for identifying bacteria, both Papanicolaou and methenamine silver stain were more cost-effective for identifying fungi. CONCLUSION: In lung FNA specimens, culture and special stains should be restricted to specimens with acute inflammation or necrosis. Gram's stain and fungal culture are insensitive and not cost-effective, and fungi are often identifiable with routine stains.

Adult↗

PubMed1994

Quantification of cell loss during bronchoalveolar lavage fluid processing. Effects of fixation and staining methods.

Discrepancies have been reported in differential cell counts according to the diverse processing methods used in bronchoalveolar lavage (BAL) fluid management. The differences have proved to be mainly the result of selective lymphocyte loss, while the exact mechanisms of the phenomenon remain controversial. Observing a similar variation in differentials from differently stained identical smears, we quantified the cell loss due to staining procedures from 45 consecutive satisfactory BAL procedures. To do this, we compared relative lymphocyte recovery on neat pooled lavage in a hemocytometer with that from smears and cytopreps fixed and stained in different ways. We found (1) A significant lymphocyte loss (p < 0.05) whatever the staining method. (2) Different methods of fixation and staining lead to considerable variation in differentials from slides otherwise identically managed. The loss is higher during air-drying fixation followed by staining with an aqueous medium such as Diff-Quik than on spray-fixed slides stained in an alcohol medium such as Papanicolaou stain. (3) The effect of lymphocyte loss on differentials is more important when the initial lymphocytosis is less than 35%, and decreases to nonsignificance when it exceeds 70%. The role of cytocentrifugation or other manipulations in cell loss probably has been overestimated because unknown effects of staining methods were also attributed to these manipulations. We suggest that lymphocyte loss could arise from poor adherence on slides, which is exacerbated during aqueous staining if no artifice (e.g., spray fixation), is used to hold them. Thus, the definition of the long-awaited standard procedure for an accurate differential count of BAL fluid must take into account fixation and staining methods.

Bronchoalveolar Lavage Fluid↗

PubMedDate not supplied

Comparison of four staining methods for detection of mast cells in equine bronchoalveolar lavage fluid.

Mast cells normally are present in equine bronchoalveolar lavage fluid (BALF), but usually represent <2% of all cells in healthy horses. An increased percentage of mast cells has been associated with airway hyperactivity and inflammatory airway diseases, but marked differences are reported between studies in normal and diseased horses. Because an abnormal mast cell count may be of clinical relevance, we compared the ability of a fast Romanowsky method to stain mast cell granules with that of 3 metachromatic stains: automated Romanowsky, May-Grünwald Giemsa, and toluidine blue stains. The BALF cells from 24 horses were studied. A differential cell count was performed blindly on 400 cells. The percentages of mast cells obtained were analyzed by means of repeated-measures analysis of variance and Fischer's PLSD test. The Bland and Altman method was used to assess agreement among stains. The mean percentage of mast cells in BALF was significantly lower with the fast Romanowsky than with the automated Romanowsky, May-Grünwald Giemsa, and toluidine blue stains. With the fast Romanowsky stain, the metachromatic granules of mast cells were not stained, and their identification was based on morphologic criteria. Toluidine blue staining allowed detection of the highest mean percentage of mast cells, but was inadequate for performing a differential cell count on other cell types. In conclusion, fast Romanosky stain may be inadequate for detection of mast cells in equine BALF, whereas automated Romanowsky, May-Grünwald Giemsa, and toluidine blue stains provide metachromatic staining of mast cell granules.

Animals↗

PubMed1980

Alcoholic Bouin fixation of insect nervous systems for Bodian silver staining. II. Modified solutions.

A previously devised synthetic equivalent of 'aged alcoholic Bouin (Duboscq-Brasil) fixative was modified in various ways to discover which of the chemical changes brought about by aging were important in improving fixation and staining. Effects were tested with ventral nerve cord ganglia of the cockroach Periplaneta americana, locust Schistocerca gregaria, and honey bee Apis mellifera. Formation of reaction products, chiefly ethyl acetate and diethoxymethane, seemed to play only a subsidiary role: neither individually appeared essential as long as a sufficient quantity of one or the other was present. In place of diethoxymethane, ethyl acetate concentration could be increased to 25% with little effect on results. Reduction in concentration of two of the original constituents, formaldehyde and ethanol, appeared to be the principal factor in improving fixation. Varying the concentration of each original constituent individually revealed that formaldehyde mainly increased glial staining, ethanol increased tissue shrinkage and reduced overall staining intensity, acetic acid improved preservation, and picric acid decreased glial staining but produced few other effects within a wide range of concentrations, though its omission seriously impaired overall preservation and staining. Varying the ethanol and acetic acid concentrations simultaneously confirmed that they acted in opposite ways. A decrease in ethanol and an increase in acetic acid both improved results. The optimum mixture, 'improved synthetic alcoholic Bouin' (40% formaldehyde 0-15: ethanol 25: acetic acid 5: ethyl acetate 5: diethoxymethane 15: picric acid 0.5: water to 100), gives better preservation and more intense staining, and formaldehyde content can be varied to give the degree of glial staining and more intense staining, and formaldehyde content can be varied to give the degree of glial staining required. Without formaldehyde glial staining is virtually eliminated, while preservation and staining of the neurons appears unaffected. This modification seems to offer a valuable advance in technique.

Acetates↗

PubMed1983

Application of dual pre-embedding stains for gonadotropins to pituitary cell monolayers with avidin-biotin (ABC) and peroxidase-antiperoxidase (PAP) complexes: light microscopic studies.

Double stains for gonadotropins and gonadotropin-releasing hormone were developed for fixed whole pituitary cells from cycling female rats. Monolayer cells were stimulated with [D-Lys6]GnRH, fixed in 2.5% glutaraldehyde, and then stained for luteinizing hormone (LH) (1:50,000-12 h) or follicle stimulating hormone (FSH) (1:60,000-12 h) and the avidin-biotin-peroxidase complex technique (ABC) with a jet-black substrate (nickel intensified diaminobenzidine-DAB). This was followed by a stain for the other gonadotropin with either ABC or peroxidase-antiperoxidase complex (PAP) techniques and amber (DAB) or red (3-amino-9-ethyl-carbazole) substrates. Additional monolayers were stimulated with biotinylated [D-Lys6]GnRH and stained with the ABC technique and the black (nickel-DAB) substrate. These monolayers were then stained immunocytochemically for LH or FSH with either ABC or PAP methods and orange or red substrates. The controls showed that the omission of the second primary antiserum abolished the stain indicating that the second staining solutions did not react with components in the first group. The addition of the second peroxidase substrate in sequence after the first stain indicated that no residual peroxidase activity remained from the first stain. Our tests also showed that saponin was not needed to aid reagent or antibody penetration. The dual stains demonstrated that 50-60% of the gonadotropes stored LH and FSH together, often in separate regions of the same cell. Some cells contained only one hormone (20-22%). The dual stains for GnRH and gonadotropins demonstrated that 80-90% of the GnRH bound cells are gonadotropes. These techniques allow a study of storage sites for multiple hormones in or on whole cells.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

PubMed1987

A combined stain for identifying epithelial cells of the gastric mucosa.

New techniques are proposed for differentiating each type of gastric epithelial cell in the same tissue section. The techniques combine the following stains: paradoxical concanavalin A staining (PCS) to identify mucous neck cells, periodic acid Schiff-concanavalin A staining to distinguish mucous neck cells from surface mucous cells, and a modified Bowie's stain to demonstrate zymogen granules of chief cells. Feulgen hydrolysis preceding the Bowie stain was found to remove most of the nonspecific coloration encountered with the original Bowie method. The results obtained by the new sequences were as follows: Feulgen hydrolysis-PCS-Bowie staining: mucous neck cells stained brown and chief cell zymogen granules deep blue. The other mucin-secreting cells remained unstained; Feulgen hydrolysis-PAS-concanavalin A-Bowie staining: mucous neck cells stained brown, zymogen granules stained deep blue to purplish blue and surface mucous cells stained purplish red.

Adult↗

PubMed1993

Immunopathological stain of lipoarabinomannan-B (LAM-B) for diagnosis of leprosy.

We developed an immunopathological staining of LAM-B antigen in formalin-fixed paraffin-embedded tissues, and compared it with, PGL-I immunostaining, Fite Faraco's stain and periodic acid carbol pararosaniline (PACPR) stain. Out of the total 28 leprosy cases, 27 were positive to LAM-B immunostaining while 23 were positive to PGL-I stain. Fite's stain was positive in 21 cases while PACPR stain was positive in 24 cases. In scrofuloderma, LAM-B antigen was observed only in the granuloma while no other positive findings were noted with other stains. Normal skin did not give any positive findings with any of the stains. Other dermatoses showed no positive findings to any of the stains tested. LAM-B staining was observed in the nerve even in the absence of bacilli in leprosy tissues. Presence of LAM-B in the cutaneous nerves is helpful in discriminating leprosy from other mycobacterioses. Considering the high sensitivity of LAM-B and the predilection of M. leprae for the nerves, we concluded that LAM-B staining can be a useful new tool in the prompt diagnosis of leprosy, especially in suspected or early cases.

Diagnosis, Differential↗

PubMed2000

Permeability of boar and bull spermatozoa to the nucleic acid stains propidium iodide or Hoechst 33258, or to eosin: accuracy in the assessment of cell viability.

This study was designed to assess whether nucleic acid stains such as propidium iodide and Hoechst 33258 and the cytosolic stain eosin identified equivalent proportions of non-viable cells. Sub-samples of boar spermatozoa stored for up to 72 h, and frozen bull spermatozoa stored in straws and thawed before staining, were exposed to either propidium iodide or Hoechst 33258 alone or in combination. Additional sub-samples were stained with eosin-nigrosin and subsequently with Giemsa. The proportion of non-viable cells identified by propidium iodide alone was equivalent to that observed when it was used in combination with the other fluorescent probe. Similar results were observed for Hoechst 33258. However, direct microscopic examination of sub-samples exposed to both stains revealed that a proportion of spermatozoa stained with propidium iodide did not incorporate Hoechst 33258. This was found consistently in boar and bull spermatozoa under the different experimental conditions used. Quantification showed that the proportion of propidium iodide-positive cells was significantly higher than Hoechst 33258-positive cells. Furthermore, the proportion of propidium iodide-positive cells was higher than cells stained with eosin, but no differences were found between the number of cells stained with Hoechst 33258 or eosin. The proportion of cells stained with propidium iodide was positively correlated with the proportion stained with either Hoechst 33258 or eosin, despite the observation that more cells incorporated propidium iodide. Taken together, these results indicate that there are differences in the ability of fluorescent probes to identify non-viable sperm cells and that this should be considered when staining protocols are used to analyse sperm viability, or when viability is used as a discriminating factor in functional studies, such as those related to acrosomal exocytosis.

Analysis of Variance↗

PubMed2000

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

The chief limitation of Ultra Fast Papanicolaou (UFP) stain, suggested by Yang and Alvarez (1995), is that, Richard Allan Haematoxylin (RA-H) and Richard Allan Cytostain (RA-C), used in the staining procedure are not universally available. It has not been established so far, whether, changes in the reagents and their proportions depending on the local availability, influence the performance of the stain. The objective of this study was to assess the feasibility and applicability of a modified UFP stain to suit our local laboratory supplies of chemical dyes, as applied to the permanent smears prepared from Fine Needle Aspiration (FNA) of breast lumps. In the present study, smears from FNA from 100 breast lumps were stained by the modified UFP stain. The modification consisted of following two changes: use of Gill's Haematoxylin instead of RAH and omission of Orange G from cytostain. Eighty Six breast aspirates were adequate for interpretation. Smears showed transparent cells with crisp nuclear features, equal to and even better than the conventional Papanicolaou stain, in a blood free background. There was an increase in total staining time by 40 seconds. We recommend the use of this modified UFP stain, only if similar reagents are being used in other laboratories. Otherwise situation specific modifications may be needed. If the UFP stain is to be used for tissues where the chances of cytoplasmic keratinization are negligible, then the use of Orange G component of the stain may become redundant.

Biopsy, Needle↗

PubMed2003

[Significance of screening by iodine staining of endoscopic examination in the area of high incidence of esophageal carcinoma].

BACKGROUND & OBJECTIVE: In order to improve early detection and visualization of esophageal premalignant lesion and carcinoma, this study was designed to explore significance of the screening by iodine staining of endoscopic examination in the area of high incidence of esophageal carcinoma. METHODS: Three screenings were completed through endoscopic examination for 3164 persons in high-risk population. Esophageal epithelium was stained with iodine in endoscopic examination. According to mucosal color and pathological result, esophageal carcinoma and premalignant lesion were confirmed. RESULTS: After staining, the normal epithelium became brown and precancerous or malignant epithelium manifested yellow with sharp margin. (1) 100% early esophageal cancer was iodine staining positive, all lesions were grade I to grade II. The incidence rates of early esophageal carcinoma and advanced esophageal carcinoma were 1.60%-4.59% and 0.29%-1.09%, respectively. Early detection rate of esophageal carcinoma was all over 75%. (2) 95.6% severe dysplasia were staining positive; 91.3% lesion were grade I and grade II in staining. The incidence rate of severe dysplasia was 4.49%-7.68%. (3) 96.6% moderate dysplasia was positive in iodine staining, 73.3% lesion were grade II and grade III. (4) 92.3% mild dysplasia was positive in iodine staining; most of lesions were grade III. (5) 0.9% esophagitis and 0.4% normal esophageal mucosal were grade I and grade II in iodine staining, most of lesion were negative or grade III. CONCLUSION: The results demonstrated that the screening with endoscopic staining examination could improve early detection of esophageal carcinoma and precancerous lesion. Iodine staining showed highly sensitivity and specificity for identifying these precancerous and early squamous cancer lesions; it may be helpful for diagnosis of early esophageal carcinoma and precancerous lesion.

Endoscopy↗

PubMed2003

Analysis of the reliability of manual and automated immunohistochemical staining procedures. A pilot study.

OBJECTIVE: To study the variation in the number of stained cells and staining intensity comparing 2 immunostainers and manual staining for estrogen receptor (ER) expression in breast carcinoma. STUDY DESIGN: In 5 cases, 15 consecutive paraffin sections were investigated after simultaneous immunohistochemical ER staining. The slides were evaluated using a CM-2 TV image analysis system (Hund, Wetzlar, Germany). One viewing field, identified around a histologic structure present on all 15 sections, was analyzed. The percentage of immunoreactive cells (PP), mean grey values of the immunopositive (GVpos.) and immunonegative nuclei (GVneg.), and immunohistochemical staining intensity (SI, defined as GVneg.-GVpos.) were calculated. RESULTS: The mean PP values were higher for immunostainers A (70.2%) and B (53.8%) than for manual staining (40.8%). The results were significantly different comparing the 2 immunostainers (P = .0143) or immunostainer A and manual staining (P < .0001). Also, the mean SI values were higher for immunostainers A (24.5 +/- 2.8% [CV]) and B (18.5 +/- 31.1%) than for manual staining (10.8 +/- 33.8%). These differences revealed statistical significance comparing the immunostainers with manual staining (.0001 < P = .0048). CONCLUSION: Our results underline the higher staining quality using immunostainers in comparison with manual staining.

Automation↗

PubMed2005

[Gimenez staining: a rapid method for initial identification of legionella pneumophila in Amoeba trophozoite].

OBJECTIVE: To establish a rapid staining method for facilitating initial identification of Legionella pneumophila in amoebal trophozoite. METHODS: Acanthamoeba polyphaga and Legionella pneumophila were co-cultured under laboratory condition. At consecutive time points during the culture, smears of the cultured products were made on glass slides for staining purposes. Different types of stainings including Gram's staining, Gimenez staining, Giemsa staining and immunofluorescence were used to determine the best method for the identification of amoebal pathogens. RESULTS: Gimenez staining technique is simpler and yields better results as compared with the other three stainings. Gimenez stain gives the best color and contrast for amoeba and amoebal Legionella Amoeba trophozoites and/or cysts showed a distinct purplish blue with amoebal Legionella in red. Amoebal Legionella can be distinguished clearly at an earlier time of co cul ture, providing a proper sensitivity. It takes only 10 minutes to finish the operation. The other techniques require the use of expensive reagents, are relatively time-consuming, and involve complex staining procedures. CONCLUSION: Gimenez staining is of value for the initial identification of amoebal pathogens, and it is suitable for laboratory diagnosis.

Amoeba↗

PubMed1976

Evaluation of bovine spermatozoal morphologic features after staining or fixation.

Two experiments were conducted to evaluate effects of 3 stains and 2 fixatives on morphologic features of bovine spermatozoa. In experiment 1, the morphologic features of acrosomes of raw and incubated, extended spermatozoa were evaluated after staining with Hancock's Blom's or Wells-Awa's stains or after fixation with buffered glutaraldehyde. Evaluations were done of stained smears by bright field microscopy and of fixed, unstained preparations, by differential interference contrast microscopy, using wet mounts. Raw semen samples from 1st ejaculates of 80 bulls were evaluated. The percentage of spermatozoa with intact acrosomes averaged 83.5% in unstained preparations fixed in glutaraldehyde, compared with averages of 68.1, 74.5, and 67.4% for smears stained with Hancock's, Blom's, or Wells-Awa's procedures (P less than 0.01). From these results, it appeared that procedures for preparing stained smears were detrimental to acrosomes. Although counts for other acrosomal abnormalities differed (P less than 0.01) in each treatment, patterns were inconsistent. With incubated, extended spermatozoa from 57 bulls, glutaraldehyde-fixed, unstained samples had more (55%) intact acrosomes (P less than 0.01) than did samples stained with Hancock's or Blom's procedures (24.0 and 34.7%, respectively, but the former were not significantly different from Wells-Awa-stained smears (49.3% intact acrosomes). In experiment 2, several morphologic characteristics of spermatozoa from 15 1st ejaculates of 7 bulls were evaluated after staining with Hancock's or Blom's stains or after fixation in buffered glutaraldehyde or buffered formal saline fixatives. Higher counts (P less than 0.01) of head abnormalities were found in wet, unstained fixed preparations (4.83, 4.47, 7.87, and 7.93% respectively, for Hancock's, Blom's, glutaraldehyde, and formol saline methods). There were more (P less than 0.05) separated heads on stained, dry smears (1.43, 1.23, 0.47, and 0.47%, respectively, for Hancock's, Blom's, glutaraldehyde, and formol saline procedures). Fixation with buffered glutaraldehyde resulted in higher counts (P less than 0.01) of proximal protoplasmic droplets (2.47, 1.03, 0.67, and 1.43%, respectively, for glutaraldehyde, Hancock's, Blom's, and formol saline procedures). Although not significant, the same trend was observed for distal protoplasmic droplets...

Acrosome↗

PubMed1982

Staining of acid-hydrolysed tissue sections with Schiff-type dye-reagents under UV rays.

This paper reports on the use of some Schiff-type dye-reagents prepared with oxalic acid and sodium thiosulphate. Some of the dyes, such as azure A, azure B, toluidine blue O, thionine, brilliant cresyl blue, and methylene violet were also prepared with oxalic acid and sodium thiosulphate but fortified with disodium hydrogen phosphate. It has been found that all these dye-reagents, barring neutral red-SO2, when used on acid-hydrolysed mammalian tissue sections under UV rays yield far better staining of the DNA-aldehyde molecules than is possible in controls stained under normal laboratory conditions. Sections stained with any of the dye-reagents under UV rays do withstand treatment in SO2 water or acid water without showing any leaching of the dye from the nuclei. Aqueous solutions of all these dyes can also be used to stain DNA-aldehyde molecules of acid-hydrolysed tissue sections. But sections stained with aqueous solutions of the azures, safranine, phenosafranine, and methylene violet when treated with SO2 water reveal considerable leaching of the dye from the nuclei. However, leaching does not occur when these dyes are used as SO2-containing dye-reagents under UV rays. It has, therefore, been concluded that in the case of staining with the azures, safranine, phenosafranine, and methylene violet, containing SO2, the reaction under UV rays is of Feulgen-type. In the case of staining with toluidine blue-SO2, thionine-SO2, and brilliant cresyl blue-SO2 under UV rays, the mechanism of staining is also of Feulgen-type but it is due to liberation of the dye molecules from their more or less colourless states that react with the DNA-aldehyde molecules producing nuclear colouration as though these dyes are in aqueous solution. This interpretation is based on the fact that sections from which RNA has been extracted selectively with cold concentrated phosphoric acid when treated with these dye-reagents under UV rays also stain DNA-phosphate groups. The increased staining intensity of the nuclei produced with the majority of the dye-reagents under UV rays as compared with control sections stained under usual laboratory conditions has been considered to be due to electronic excitation of the dye molecules thus facilitating binding between more molecules of DNA-aldehyde and of dye. Possible significance of these findings has been discussed.

Animals↗

PubMed1983

Influence of staining on fast automated cell segmentation, feature extraction and cell image analysis.

FAZYTAN, a system for fast automated cell segmentation, cell image analysis and extraction of nuclear features, was used to analyze cervical cell images variously stained by the conventional Papanicolaou stain, the new Papanicolaou stain and hematoxylin and thionin only; the last two dyes are used as the nuclear stains in the two versions of the Papanicolaou stain. Other dyes were also tried in cell classification experiments. All cell images in the variously stained samples could be described by the same nuclear features as had been adapted for the discrimination of conventional-Papanicolaou-stained cells. Variances were lower for thionin-stained cells as compared with hematoxylin-stained cells. By application of spectrophotometry, it was confirmed that the spectra of the cytoplasmic counterstains are superimposed on those of the nuclear stains. It appears that a variety of dyes are suitable as cytologic stains for cell classification by the FAZYTAN system, provided that they achieve sufficiently strong nuclear-cytoplasmic contrast by precisely delineating the chromatin texture.

Cell Nucleus↗

PubMed1983

[Staining behavior and applicability of spectrally pure Capriblue GN, Stella Blue, Oxonin and Punky Blue].

The staining properties of 4 spectral pure derivates of Phenoxazin--Capriblue GN, Stella Blue, Oxonin, and Punky Blue--were investigated using human and animal tissues. Punky Blue and Stella Blue are newly synthesized derivates. A simplified staining technique was developed based on conventional fixing and mounting methods of histological materials. Punky Blue is a particularly contrasting metachromatic nucleus stain, and produces at pH = 4 and a staining period of 5 min a differentiated cell picture, the colours of which are comparable to conventional stains. The Oxonin-stain produces good contrasts after 2 min. Capriblue GN is only limited use as one-component tissue stain, due of its low metachromatic characteristics and Stella Blue, on its own, does not provide sufficient contract. The influence of electrolytes and hydrogen ions on the staining mechanisms will be discussed. An increase of hydrogen ions is used to control the selective staining of amphoteric biopolymers. The possible binding mechanisms of stains used are discussed in the light of their staining properties.

Chemical Phenomena↗