von Ruhland C J
Electron and Light Microscopy Facility, Central Biotechnology Services, School of Medicine, Cardiff University, Cardiff, UK.
J Microsc. 2025 Jan;297(1):13-17. doi: 10.1111/jmi.13359. Epub 2024 Sep 16.
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.
在将组织切片加工成用于光学和电子显微镜检查的树脂的过程中，由于各种组织成分的收缩差异，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本切片。为了缓解这个问题，人们设计了一些技术，以确保样本在树脂最终固化/聚合过程中保持平整。对于丙烯酸树脂包埋，必须排除氧气，因为它会抑制聚合反应，而设计用于环氧树脂包埋的方法通常不合适。本文介绍了由聚酯薄膜制备的气密扁平包埋室的制备和使用方法，它为以前提倡用于此目的的由聚对苯二甲酸乙二酯盖玻片或氟化乙烯丙烯共聚物薄膜制成的包埋室提供了一种廉价、技术上更简单且通用的替代方案。通俗描述：在将组织切片加工成用于光学和电子显微镜检查的树脂的过程中，由于各种组织成分的收缩差异，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。对于丙烯酸树脂包埋，必须排除氧气，因为它会抑制聚合反应，而设计用于环氧树脂扁平包埋的方法通常不合适。以前有人提倡使用由聚对苯二甲酸乙二酯盖玻片或由涂有聚四氟乙烯的载玻片、聚碳酸酯薄膜垫圈和氟化乙烯丙烯共聚物薄膜组合制成的气密扁平包埋室来达到此目的。聚对苯二甲酸乙二酯盖玻片价格昂贵，尺寸限制为22毫米×60毫米，而替代方法技术复杂。聚酯薄膜以210毫米×297毫米的片材形式商业出售，价格约为聚对苯二甲酸乙二酯盖玻片的1/20，不到氟化乙烯丙烯共聚物薄膜价格的一半。本文介绍了由聚酯薄膜、载玻片和双面胶带制成的包埋室的制备和使用方法，作为聚对苯二甲酸乙二酯盖玻片和氟化乙烯丙烯共聚物薄膜方法的一种技术上更简单、廉价且通用的替代方案。
