Standardized Methyl Green—Pyronin Y procedures using pure dyes

Høyer, P. E.; Lyon, H.; Jakobsen, Palle; Andersen, A. P.

doi:10.1007/bf01675361

Cited by 16 publications

(6 citation statements)

References 10 publications

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“…Methyl green-pyronin staining was performed according to the method of Hayer et al (1986). Sections prepared with these stains were examined under the light microscope.…”

Section: Methodsmentioning

confidence: 99%

Neurodegeneration with Tau Accumulation in a Transgenic Mouse Expressing V337M Human Tau

Tanemura

Murayama

Akagi³

et al. 2002

J. Neurosci.

220

118

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Formation of neurofibrillary tangles (NFTs) is a common neuropathological feature found in several neurodegenerative diseases, including Alzheimer's disease. We have developed a transgenic (Tg) mouse expressing mutant human tau (V337M), derived from frontotemporal dementia parkinsonism-17. V337M Tg mice revealed tau aggregations in the hippocampus, which fulfills the histological criteria for NFTs in human neurodegenerative diseases. Concurrent with the accumulation of RNA and phosphorylated tau, neurons exhibited morphological characteristics of degenerating neurons, which include a loss of microtubules, accumulation of ribosomes, plasma and nuclear membrane ruffling, and swelling of the Golgi network. Thus, mutant tau induces neuronal degeneration associated with the accumulation of RNA and phosphorylated tau. The functional consequences of this neuronal degeneration was evidenced by the reduction of hippocampal neural activity and behavioral abnormality in Tg mice.

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“…Methyl green-pyronin staining was performed according to the method of Hayer et al (1986). Sections prepared with these stains were examined under the light microscope.…”

Section: Methodsmentioning

confidence: 99%

Neurodegeneration with Tau Accumulation in a Transgenic Mouse Expressing V337M Human Tau

Tanemura

Murayama

Akagi³

et al. 2002

J. Neurosci.

220

118

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“…Comparison of the standardized simultaneous and sequential procedures described in Hoyer et al (1986) using the colour TV-image analysis as described in the present paper unequivocally shows that even though pleasant appearances may be obtained with the Pyronin/Methyl Green sequence as evaluated by visual examination, the results obtained with the sequential methods do not show a simple stoichiometric relationship to the substrate but instead represent procedures with an element of 'differentiation'. As a result we can no longer recommend the use of the sequential methods for quantitative purposes.…”

Section: Discussionmentioning

confidence: 84%

“…The following staining solutions were prepared according to Hoyer et al (1986) Graumann (1953) using pure Pararosanilin hydrochloride (Serva GmbH, Germany, batch no. P-8101).…”

Section: Dyes and Staining Solutionsmentioning

confidence: 99%

The correlation between uptake of Methyl Green and Feulgen staining intensity of cell nuclei. An image analysis study

1989

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Paraffin sections of rat tissue fixed in either formaldehyde solution (3.6% w/v) or in Carnoy's fluid were stained using standardized Methyl Green-Pyronin procedures with the dyes used either simultaneously or in sequence. The sections were evaluated for the uptake of the two dyes by cell nuclei, nucleoli and cytoplasm using colour TV-image analysis. The parameters measured were integrated optical density and the surface area of the object. The sections were then destained and a Feulgen reaction was performed. The coordinates of the cells measured after the simultaneous Methyl Green-Pyronin method were stored in the computer, making it possible to measure the same cells in the Feulgen-restained sections. Image analysis gave results which invalidate the sequential methods as opposed to the simultaneous method. Mean optical densities were significantly increased for both dyes with the simultaneous method after formaldehyde fixation as compared to Carnoy fixation. The quantitative correlation of Methyl Green and DNA in the simultaneous technique was found to parallel exactly that of the Feulgen stain. In conclusion, the simultaneous Methyl Green-Pyronin technique is recommended while the sequential methods seem to be of less value.

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“…229-34) containing specified amounts of various molecules in a suitable matrix (van der Ploeg & van Duijn, 1964;van Duijn, 1976;van Duijn & van der Ploeg, 1980) has given much valuable information for achieving these goals. Examples of standardization involving both the dyes and their applications are the standard Romanowsky-Giemsa stain (see Part 4 of this review) and the Methyl Green-Pyronin Y method (Hoyer et al, 1986). For further references see, for instance, Schulte et al (1992).…”

Section: Standardization Of Methodsmentioning

confidence: 99%

Standardization of reagents and methods used in cytological and histological practice with emphasis on dyes, stains and chromogenic reagents

et al. 1994

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The need for the standardization of reagents and methods used in the histology laboratory is demonstrated. After definitions of dyes, stains, and chromogenic reagents, existing standards and standards organizations are discussed. This is followed by practical instructions on how to standardize dyes and stains through the preparation of reference materials and the development of chromatographic methods. An overview is presented of the problems concerned with standardization of the Romanowsky-Giemsa stain for cytological and histological application. Finally, the problem of how to convince routine dye and stain users of the need for standardization in their histology laboratories is discussed.

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Standardized Methyl Green—Pyronin Y procedures using pure dyes

Cited by 16 publications

References 10 publications

Neurodegeneration with Tau Accumulation in a Transgenic Mouse Expressing V337M Human Tau

Neurodegeneration with Tau Accumulation in a Transgenic Mouse Expressing V337M Human Tau

The correlation between uptake of Methyl Green and Feulgen staining intensity of cell nuclei. An image analysis study

Standardization of reagents and methods used in cytological and histological practice with emphasis on dyes, stains and chromogenic reagents

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