Heck Christian T, Volkmann Gwyneth, Woodward Holly N
Department of Biomedical Sciences, Oklahoma State University Center for Health Sciences, Tulsa, OK, United States of America.
PeerJ. 2020 Dec 15;8:e10495. doi: 10.7717/peerj.10495. eCollection 2020.
Histological examination of bone microstructure provides insight into extant and extinct vertebrate physiology. Fossil specimens sampled for histological examination are typically first embedded in an inexpensive polyester resin and then cut into thin sections, mounted on slides, and polished for viewing. Modern undecalcified bone is chemically processed prior to embedding in plastic resin, sectioning, mounting, and polishing. Conversely, small fossil material and modern undecalcified bone are typically embedded in higher priced epoxy resin because these specimen types require final sections near or below 100 µm thick. Anecdotal evidence suggests thin sections made of polyester resin embedded material polished thinner than 100 µm increases likelihood of sample peeling, material loss, and is unsuitable for modern tissue and small fossil material. To test this assertion, a sample of modern bones and fossil bones, teeth, and scales were embedded in either polyester resin or epoxy resin. Embedded specimens were sectioned and mounted following standard published protocol. Thin sections were ground on a lapidary wheel using decreasing grit sizes until tissue microstructure was completely discernible when viewed under a polarizing light microscope. Additionally, eight prepared thin sections (four from polyester resin embedded specimens and four from epoxy resin embedded specimens) were continuously ground on a lapidary wheel using 600 grit carbide paper until peeling occurred or material integrity was lost. Slide thickness when peeling occurred was measured for comparing slide thickness when specimen integrity was lost between the two resin types. Final slide thickness ranged from 38 µm to 247 µm when tissue was identifiable using a polarizing microscope. Finished slide thickness varied between resin types despite similar tissue visibility. However, finished slide thickness appears more dependent on hard tissue composition than resin type. Additionally, we did not find a difference of slide thickness when material was lost between resin types. The results of this preliminary study suggest that polyester resins can be used for embedding undecalcified modern hard tissues and fossilized hard tissues without loss of tissue visibility or material integrity, at least in the short term.
对骨微观结构进行组织学检查有助于深入了解现存和已灭绝脊椎动物的生理机能。用于组织学检查的化石标本通常首先被嵌入一种价格低廉的聚酯树脂中,然后切成薄片,安装在载玻片上,并进行抛光以便观察。现代未脱钙骨在嵌入塑料树脂、切片、安装和抛光之前需要进行化学处理。相反,小型化石材料和现代未脱钙骨通常嵌入价格较高的环氧树脂中,因为这些标本类型需要最终切片厚度接近或低于100微米。有传闻证据表明,由聚酯树脂嵌入材料制成的薄片被抛光至厚度小于100微米时,样品剥落、材料损失的可能性会增加,并且不适用于现代组织和小型化石材料。为了验证这一说法,将一组现代骨骼、化石骨骼、牙齿和鳞片样本分别嵌入聚酯树脂或环氧树脂中。按照已发表的标准方案对嵌入的标本进行切片和安装。使用粒度逐渐减小的磨盘在切片机上研磨薄片,直到在偏光显微镜下观察时组织微观结构完全清晰可见。此外,使用600目硬质合金砂纸在切片机上对八个制备好的薄片(四个来自聚酯树脂嵌入标本,四个来自环氧树脂嵌入标本)持续研磨,直到出现剥落或材料完整性丧失。测量出现剥落时的载玻片厚度,以比较两种树脂类型在材料完整性丧失时的载玻片厚度。当使用偏光显微镜能够识别组织时,最终载玻片厚度范围为38微米至247微米。尽管组织可见性相似,但最终载玻片厚度在树脂类型之间存在差异。然而,最终载玻片厚度似乎更多地取决于硬组织成分而非树脂类型。此外,我们没有发现两种树脂类型在材料损失时载玻片厚度存在差异。这项初步研究的结果表明,聚酯树脂可用于嵌入未脱钙的现代硬组织和化石硬组织,至少在短期内不会损失组织可见性或材料完整性。