Use of Melinex film for flat embedding tissue sections in LR White

IF 1.5 4区 工程技术 Q3 MICROSCOPY Journal of microscopy Pub Date : 2024-09-16 DOI:10.1111/jmi.13359
C. J. von Ruhland
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Abstract

Tissue slices can undergo distortions during processing into resin for light and electron microscopy as a result of differential shrinkage of the various tissue components, and this may necessitate removal of a considerable amount of material from the final resin-embedded tissue block to ensure production of complete sections of the sample. To mitigate this problem, a number of techniques have been devised that ensure the sample is held flat during the final curing/polymerisation of the resin. For embedding in acrylic resins, oxygen must be excluded as it inhibits polymerisation, and methods devised for epoxy resin embedding are generally unsuitable. The method describes the preparation and use of air-tight flat-embedding chambers prepared from Melinex film and provides an inexpensive, technically simpler, and versatile alternative to chambers formed from either Thermanox coverslips or Aclar films that have previously been advocated for such purposes.

Lay description: Tissue slices can undergo distortions during processing into resin for light and electron microscopy as a result of differential shrinkage of the various tissue components. Such distortions may necessitate removal of a considerable amount of material to ensure production of complete sections of the sample. For embedding in acrylic resins, oxygen must be excluded as it inhibits polymerisation, and methods devised for epoxy resin flat-embedding are generally unsuitable. Air-tight flat-embedding chambers prepared from either Thermanox coverslips, or a combination of PTFE-coated glass slides, polycarbonate film gaskets, and Aclar film have been advocated for such purposes. Thermanox coverslips are expensive and limited in size to 22 mm × 60 mm, and the alternative method is technically complicated. Melinex film is commercially available as 210 mm × 297 mm sheets and is approximately 1/20th the price of Thermanox and less than half the price of Aclar film. The method describes the preparation and use of embedding chambers made from Melinex film, glass slides and double-sided adhesive tape as a technically simpler, inexpensive and versatile alternative to both Thermanox coverslips and the Aclar film method.

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使用 Melinex 薄膜将组织切片平铺包埋在 LR White 中。
在将组织切片加工成用于光镜和电子显微镜观察的树脂过程中,由于各种组织成分的收缩程度不同,组织切片可能会发生变形,这可能需要从最终的树脂包埋组织块中去除大量材料,以确保制作出完整的样品切片。为了缓解这一问题,我们设计了一系列技术来确保样本在树脂最终固化/聚合过程中保持平整。对于丙烯酸树脂的包埋,必须排除氧气,因为氧气会抑制聚合,而环氧树脂的包埋方法一般都不适用。该方法描述了用 Melinex 薄膜制备和使用气密性平面包埋室的方法,与以前提倡的用 Thermanox 盖玻片或 Aclar 薄膜制备的包埋室相比,该方法成本低廉、技术简单、用途广泛。铺设说明:在将组织切片加工成用于光镜和电子显微镜的树脂过程中,由于各种组织成分的收缩程度不同,组织切片可能会发生变形。这种变形可能需要去除大量材料,以确保制作出完整的样本切片。在丙烯酸树脂中进行包埋时,必须排除氧气,因为氧气会抑制聚合作用,而为环氧树脂平面包埋设计的方法一般都不适用。为此,有人主张使用 Thermanox 盖玻片或聚四氟乙烯涂层玻璃载玻片、聚碳酸酯薄膜垫圈和 Aclar 薄膜组合制作气密性平片包埋室。Thermanox 盖玻片价格昂贵,尺寸仅限于 22 毫米 × 60 毫米,而且替代方法在技术上比较复杂。Melinex 胶片在市场上有售,规格为 210 毫米 × 297 毫米,价格约为 Thermanox 的 1/20,不到 Aclar 胶片的一半。该方法介绍了如何制备和使用由 Melinex 薄膜、玻璃载玻片和双面胶带制成的包埋室,作为 Thermanox 盖玻片和 Aclar 薄膜方法的一种技术简单、价格低廉且用途广泛的替代方法。
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来源期刊
Journal of microscopy
Journal of microscopy 工程技术-显微镜技术
CiteScore
4.30
自引率
5.00%
发文量
83
审稿时长
1 months
期刊介绍: The Journal of Microscopy is the oldest journal dedicated to the science of microscopy and the only peer-reviewed publication of the Royal Microscopical Society. It publishes papers that report on the very latest developments in microscopy such as advances in microscopy techniques or novel areas of application. The Journal does not seek to publish routine applications of microscopy or specimen preparation even though the submission may otherwise have a high scientific merit. The scope covers research in the physical and biological sciences and covers imaging methods using light, electrons, X-rays and other radiations as well as atomic force and near field techniques. Interdisciplinary research is welcome. Papers pertaining to microscopy are also welcomed on optical theory, spectroscopy, novel specimen preparation and manipulation methods and image recording, processing and analysis including dynamic analysis of living specimens. Publication types include full papers, hot topic fast tracked communications and review articles. Authors considering submitting a review article should contact the editorial office first.
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