Rapid demonstration of nucleic acids using "oxidised" gallocyanin and chromic potassium sulphate: methods and applications.

Husain, O. A. N.; Watts, K. C.

doi:10.1136/jcp.37.1.99

Cited by 13 publications

(8 citation statements)

References 6 publications

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“…Enzymatic digestion of DNA was carried out with DNAse (Boehringer, Table 1. Staining protocol for the Gallocyanin chromalum stain after Husain and Watts (1984) followed by various posttreatment procedures.…”

Section: Staining Proceduresmentioning

confidence: 99%

“…The Husain and Watts stain was prepared according to Husain and Watts (1984) from a batch of Gallocyanin chloride (Polysciences, batch no. S-61-G; C.I.…”

Section: Stainsmentioning

confidence: 99%

“…The situation has, however, changed completely with the advent of modified Gallocyanin chromalum stains with considerably shorter staining times (Eason & Tucker, 1979;Husain & Watts, 1984). Staining times of less than one hour were acceptable also for quick routine staining in diagnostic cytology.…”

Section: Introductionmentioning

confidence: 98%

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Gallocyanin chromalum as a nuclear stain in cytology. I. A cytophotometric comparison of the Husain-Watts Gallocyanin chromalum staining protocol with the Feulgen procedure

1991

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In the present study, the staining characteristics of the Gallocyanin chromalum technique devised by Husain and Watts are compared with the Feulgen reaction. Liver imprints, blood smears, and cervical smears were fixed in ethanol and stained with either the Husain and Watts Gallocyanin chromalum reagent or the Feulgen-Schiff reagent. The slides were then post-treated with 70% ethanol-HCl pH 1.0, or with phosphotungstic acid for 0.5-30 min. The integrated optical density of cell nuclei was measured with a VIDAS image analyzer. In the material stained with the Husain and Watts procedure, some Gallocyanin chromalum was removed from the nuclei in the early phase (5 min) of all the post-treatment steps, followed by a plateau phase where the integrated optical density remained constant for 30 min. In this phase, the nuclear absorbance was highly reproducible and of the same size regardless of the post-treatment. Both the Husain and Watts procedure and the Feulgen-reaction gave quantitative staining of DNA. The Gallocyanin chromalum stain after Husain and Watts is a quick staining procedure for quantitative evaluation of DNA in cytological material. Proper rinsing of the slides is necessary for a good reproducibility of results.

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Section: Staining Proceduresmentioning

confidence: 99%

“…The Husain and Watts stain was prepared according to Husain and Watts (1984) from a batch of Gallocyanin chloride (Polysciences, batch no. S-61-G; C.I.…”

Section: Stainsmentioning

confidence: 99%

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Gallocyanin chromalum as a nuclear stain in cytology. I. A cytophotometric comparison of the Husain-Watts Gallocyanin chromalum staining protocol with the Feulgen procedure

1991

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“…The overall texture measurement from an object will be made up from contributions from density changes at bounda- Image Data Base Collection The object images used for the experimental investigations were obtained from 20 cervical cytology scrape specimens with varying cytological grades. Slides were prepared using the suspension/monolayering/gallocyanin chrome alum method developed for use with the CERVIFIP system (11,22). The CERVlFIP system program was then set up to determine the positions of high-DNA objects detected as suspect cells, using a previously optimised classifier for abnormal cell detection and artifact rejection.…”

Section: Texture Features Investigatedmentioning

confidence: 99%

Interval‐coded texture features for artifact rejection in automated cervical cytology

et al. 1988

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In order to improve the separation between abnormal cells and noncellular artifacts in the CERVIFIP automated cervical cytology prescreening system, 22 different object texture features were investigated. The features were all statistical parameters of the pixel density histograms or one-dimensional filtered values of central and border regions of the object images. The features were calculated for 231 images (100 cells and 131 artifacts) detected as Suspect Cells by the current CERVIFIP and were then tested in hierarchical and linear discriminant classifiers. After selecting the two best features for use in a hierarchical classifier, 83% correct classification was achieved. One of these features was specifically designed to remove poorly focused objects. With maximum likelihood discrimination using all 22 features, an overall correct classification rate of 90% was obtained. Key terms: Image analysis, pattern recognition features, automated prescreening, cewical cytology chromatin, texture, artifactsThe problem of rejecting noncellular artifacts is one of very great practical importance in automated cytometry. For example, a major difficulty in cervical cytology automation is that of rejecting false-alarm signals ("false positives") in normal specimens caused by artifacts such as overlapping cell pairs, leukocyte clusters, dust particles, etc. Already, several techniques have been developed for this purpose based upon the analysis of global object features (e.g. area, density) (14,19), object shape (6,14,16-18,24,25), object texture (61, and object colour (6). Nevertheless, it has been found in system trials that a significant number of artifacts are not detected using classification based upon these features, so that falsepositive objects are still a major factor in specimen misclassification. Improvements in features for artifact rejection should therefore result in an increased eficiency of such systems.The analysis of cellular texture is one of the major areas of research in the extraction of information from cell images. Several different approaches have emerged, including the use of co-occurence (transition) matices (23), grain analysis (13), one-and two-dimensional filtering (lo), run-length texture analysis (8), and statistical parameters (1). In quantitative cytology, the analysis of nuclear texture has been particularly successful in separating normal and malignant specimens, even in apparently normal cells, (3,4), because texture measurements on an appropriately prepared and stained cell nucleus reflect the local chromatin arrangement, which is one of the most powerful indicators of cell function and hence of cell pathology.One type of instrumentation that has found application in the field of automated cytometry is that of the continuous motion imaging (CMD system, in which the specimen is moved continuously under a one-dimensional scanner (e.g., linear CCD diode array) (2,12,15).These systems possess several desirable properties for cervical cytology automation; they are fast, and they have potentia...

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“…Vasculature devoid of blood may be filled with a radiopaque suspension of lead, bismuth, or barium, which solidifies before scanning (e.g., "microfil") to opacify the microvasculature (Proweller et al 2007;Degenhardt et al 2010). Chromium (Z = 24) contrast is imparted to nuclei using the histological stain gallocyanin-chromalum, first described by Husain and Watts (1984) and further described for use in micro-CT imaging by Metscher (2009b). Osmium is recognized to differentially stain tissues with high lipid content, including white and brown adipose tissue, and contrasts the lipoproteins of most bilayered cell and organelle membranes (Hayat 1970).…”

mentioning

confidence: 99%

A Correlative Method for Imaging Identical Regions of Samples by Micro-CT, Light Microscopy, and Electron Microscopy

Sengle

Tufa

Sakai

et al. 2012

J Histochem Cytochem.

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We present a method in which a precise region of interest within an intact organism is spatially mapped in three dimensions by non-invasive micro-computed X-ray tomography (micro-CT), then further evaluated by light microscopy (LM) and transmission electron microscopy (TEM). Tissues are prepared as if for TEM including osmium fixation, which imparts soft tissue contrast in the micro-CT due to its strong X-ray attenuation. This method may therefore be applied to embedded, archived TEM samples. Upon selection of a two-dimensional (2-D) projection from a region of interest (ROI) within the three-dimensional volume, the epoxy-embedded sample is oriented for microtomy so that the sectioning plane is aligned with the micro-CT projection. Registration is verified by overlaying LM images with 2-D micro-CT projections. Structures that are poorly resolved in the micro-CT may be evaluated at TEM resolution by observing the next serial ultrathin section, thereby accessing the same ROI by all three imaging techniques. We compare white adipose tissue within the forelimbs of mice harboring a lipid-altering mutation with their littermate controls. We demonstrate that individual osmium-stained lipid droplets as small as 15 µm and separated by as little as 35 µm may be discerned as separate entities in the micro-CT, validating this to be a high-resolution, non-destructive technique for evaluation of fat content.

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Rapid demonstration of nucleic acids using "oxidised" gallocyanin and chromic potassium sulphate: methods and applications.

Cited by 13 publications

References 6 publications

Gallocyanin chromalum as a nuclear stain in cytology. I. A cytophotometric comparison of the Husain-Watts Gallocyanin chromalum staining protocol with the Feulgen procedure

Gallocyanin chromalum as a nuclear stain in cytology. I. A cytophotometric comparison of the Husain-Watts Gallocyanin chromalum staining protocol with the Feulgen procedure

Interval‐coded texture features for artifact rejection in automated cervical cytology

A Correlative Method for Imaging Identical Regions of Samples by Micro-CT, Light Microscopy, and Electron Microscopy

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