Use of Semithin Sections Embedded in a Water-Miscible Methacrylate for Light Microscopy of Central Nerve Tissues

Nagato, Yasukazu; Sekiguchi, Masaki; Kushida, Tsuyuka; Kushida, Hiroshi; Shimai, Kazuyo

doi:10.2535/ofaj1936.66.2-3_145

Cited by 4 publications

(4 citation statements)

References 16 publications

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“…This method improved the formulation of GMA, Quetol 523 and MMA by employing HPMA instead of GMA. A low distortion and high stainability were also characteristic of previous GMA-Quetol 523-MMA (Kushida et al, 1981) and HPMA-Quetol 523-MMA embedded tissue sections, but the latter had several advantages over the former for the light microscopy of nerve tissues (Nagato et al, 1989). Quetol 523 and MMA reacted with HPMA and became an integral part of the polymerized system without a cross-linker.…”

Section: Discussionmentioning

confidence: 88%

“…Thus, structural details that are not apparent in sections of greater than 2.0 im in thickness may be visible in thinner sections. Nagato et al (1989) reported that the best results were obtained with tissue sections of 1.5 [im in thickness when observing the structure of nerve cells.…”

mentioning

confidence: 99%

“…The use of HPMA instead of GMA was devised for easier embedding, infiltration, sectioning and staining than in the case of GMAQuetol 523-MMA embedding. In addition, useful staining and related procedures were also reported for central nervous tissues by light microscopy (Nagato et al, 1989). This paper describes the application of useful staining, enzyme and immunohistochemical procedures employing a mixture of HPMA.…”

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confidence: 99%

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Staining of Semithin Tissue Sections Embedded in HPMA, Quetol 523 and MMA

Iijima

Nagato

Kushida

1992

Okajimas Folia Anatomica Japonica

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Summary: Various tissues fixed in a mixture of formaldehyde and glutaraldehyde, and embedded in an improved 2-hydroxypropyl methacrylate mixture were employed for studying the fine structures of cells and tissues by light microscopy. The embedding mixture contained Quetol 523 and methyl methacrylate as a plasticizer without a cross-linker. The catalyst was QCU-1. The mixture had a low viscosity, was easy to handle and penetrated readily and completely into the specimen, been employed with such sections and those reported here are hematoxylin-eosin, Azan and PAS. There was excellent preservation of alkaline phosphatase activity. A method of poststaining immunoperoxidase labeling was also applied to the mouse pancreas and examples of staining with insulin are included.

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Section: Discussionmentioning

confidence: 88%

mentioning

confidence: 99%

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confidence: 99%

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Staining of Semithin Tissue Sections Embedded in HPMA, Quetol 523 and MMA

Iijima

Nagato

Kushida

1992

Okajimas Folia Anatomica Japonica

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“…They were immersed in a staining solution of hematoxylineosin or toluidine blue. The sections require longer staining times than semithin sections used for light microscopy (Nagato et al , 1989, Iijima et al , 1992. After staining, the grids were carefully removed from the polyethylene tubing and placed on the slide glass for examination and photography.…”

Section: Light Microscopymentioning

confidence: 99%

Correlative Light and Electron Microscopy of the Same Sections Embedded in HPMA, Quetol 523 and MMA

Kushida

Nagato

Iijima

et al. 1993

Okajimas Folia Anatomica Japonica

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Semithin sections, cut from tissues stained with acid and basic dyes after embedding in 2-hydroxypropyl methacrylate, Quetol 523 and methyl methacrylate, showed cytoplasmic components at a high resolution by light microscopy. These same sections could then be viewed, after osmium tetroxide, uranyl and lead staining, by the electron microscope. These sections had a number of inherent advantages: they could be observed with a light microscope; they facilitated analysis of cellular structures in the identical sites, and they were frequently the optimum thickness to provide three-dimensional information. We clearly established the structural detail of this same-section correlative light-electron microscopy approach by showing that the coloured materials observed in such sections of cells followed the distribution of fine structures within the same sections as determined by electron microscopy. In some instances the fidelity of the correlation between the distribution of the coloured area and cytoplasmic components in identical cells of the same section revealed significant details which could not visualized in thin sections. This technique, therefore, provided a simple and useful solution to many problems that require the localization of cellular components in identical cells selected previously by light microscopy.

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Observation on the cycle of the seminiferous epithelium in the Japanese macaque (Macaca fuscata) using semithin sections

1994

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ABSTRACT. The stages of the cycle of the seminiferous epithelium in the Japanese macaque are investigated using testes fixed by a mixture of formaldehyde and glutaraldehyde containing picric acid and embedded in a methacrylate resin, Queto1523M. Sections, 1.0-2.0/zm in thickness, were cut with glass knives and stained with periodic acid-Schiff (PAS) and hernatoxylin. Sections from such resin blocks illustrated cellular detail without structural distortion during the polymerization process. Furthermore, staining affinity with PAS and hematoxylin was excellent. In stained sections, typical germ cell associations were described, based on the nuclear morphology of type A (dark and pale) spermatogonium, type B spermatogonium, various developmental stages of primary spermatocytes during meiosis, and the development of the acrosomic system. In the Japanese macaque, two different steps of spermatids (steps 3 and 4) were constantly seen in the same area of the tubular epithelium during stage III. Therefore, a classification into ten stages is proposed for the cycle in this species. Additional characteristics are described based on the observation of the seminiferous epithelium using semithin sections.

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Use of Semithin Sections Embedded in a Water-Miscible Methacrylate for Light Microscopy of Central Nerve Tissues

Cited by 4 publications

References 16 publications

Staining of Semithin Tissue Sections Embedded in HPMA, Quetol 523 and MMA

Staining of Semithin Tissue Sections Embedded in HPMA, Quetol 523 and MMA

Correlative Light and Electron Microscopy of the Same Sections Embedded in HPMA, Quetol 523 and MMA

Observation on the cycle of the seminiferous epithelium in the Japanese macaque (Macaca fuscata) using semithin sections

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