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A Paraffin Microtomy Method for Improved and Efficient Production of Standardized Plastic Microfibers
Environmental Toxicology and Chemistry ( IF 4.1 ) Pub Date : 2021-09-22 , DOI: 10.1002/etc.5216
Christine M Knauss 1 , Christopher F Dungan 2 , Stuart A Lehmann 2
Affiliation  

Microfibers are one of the most abundant microplastic particle types found in the environment, where they cause negative impacts on organisms and possibly on human health. Microfibers should be included in a wide range of laboratory studies; however, microfibers for scientific studies are not commercially available. Current methods to make microfibers generally create particles with large size ranges and poor precision, and efficient production of particles ≤100 µm is difficult. Laboratory studies of the biological and toxicological effects and chemical interactions of microfibers require uniform, small microfibers in sufficient numbers for environmentally relevant experiments. We developed a novel fiber embedding technique and modified a seminal cryomicrotomy method to produce precise microfibers in quantities suitable for environmentally relevant concentrations. Polyethylene terephthalate (PET) and nylon fibers were strategically wound onto a spindle, embedded in paraffin wax, and sectioned using a standard paraffin microtome. After processing with a suitable organic solvent to remove the wax, microfiber size distributions were assessed. The small microfibers (10–42 µm) were accurate to the target lengths with excellent precision and a production rate ≥13.5 times higher than previous methods. As a proof of application, three lengths of manufactured PET fibers were stained with Nile red and exposed to eastern oyster larvae (Crassostrea virginica) for 24 h. Larvae ingested the smaller fiber lengths (14 and 28 µm), and the Nile red–stained fibers were visible and distinguishable in the guts of the larvae. This experiment was the first to demonstrate ingestion of plastic particles other than microspheres by oyster larvae. The present method facilitates the use of small microfibers in laboratory experiments, allowing for a more complete understanding of microplastic effects in the environment. Environ Toxicol Chem 2022;41:944–953. © 2021 SETAC

中文翻译:

改进和高效生产标准化塑料微纤维的石蜡切片法

微纤维是环境中发现的最丰富的微塑料颗粒类型之一,它们对生物体甚至可能对人类健康造成负面影响。微纤维应包括在广泛的实验室研究中;然而,用于科学研究的超细纤维在市场上是不可购得的。当前制造微纤维的方法通常会产生尺寸范围大且精度差的颗粒,并且难以有效生产≤100 µm的颗粒。对微纤维的生物和毒理学效应以及化学相互作用的实验室研究需要足够数量的均匀、小微纤维来进行环境相关实验。我们开发了一种新的纤维嵌入技术并修改了精液冷冻切片法,以生产适合环境相关浓度的精确微纤维。聚对苯二甲酸乙二醇酯 (PET) 和尼龙纤维被战略性地缠绕在纺锤上,嵌入石蜡中,并使用标准石蜡切片机切片。在用合适的有机溶剂处理以去除蜡后,评估微纤维尺寸分布。小微纤维 (10–42 µm) 精确到目标长度,具有出色的精度和比以前方法高 13.5 倍的生产率。作为应用证明,三段人造 PET 纤维被尼罗红染色并暴露于东部牡蛎幼虫(聚对苯二甲酸乙二醇酯 (PET) 和尼龙纤维被战略性地缠绕在纺锤上,嵌入石蜡中,并使用标准石蜡切片机切片。在用合适的有机溶剂处理以去除蜡后,评估微纤维尺寸分布。小微纤维 (10–42 µm) 精确到目标长度,具有出色的精度和比以前方法高 13.5 倍的生产率。作为应用证明,三段人造 PET 纤维被尼罗红染色并暴露于东部牡蛎幼虫(聚对苯二甲酸乙二醇酯 (PET) 和尼龙纤维被战略性地缠绕在纺锤上,嵌入石蜡中,并使用标准石蜡切片机切片。在用合适的有机溶剂处理以去除蜡后,评估微纤维尺寸分布。小微纤维 (10–42 µm) 精确到目标长度,具有出色的精度和比以前方法高 13.5 倍的生产率。作为应用证明,三段人造 PET 纤维被尼罗红染色并暴露于东部牡蛎幼虫(小微纤维 (10–42 µm) 精确到目标长度,具有出色的精度和比以前方法高 13.5 倍的生产率。作为应用证明,三段人造 PET 纤维被尼罗红染色并暴露于东部牡蛎幼虫(小微纤维 (10–42 µm) 精确到目标长度,具有出色的精度和比以前方法高 13.5 倍的生产率。作为应用证明,三段人造 PET 纤维被尼罗红染色并暴露于东部牡蛎幼虫(牡蛎) 24 小时。幼虫摄入的纤维长度较短(14 和 28 µm),尼罗红染色的纤维在幼虫的内脏中可见且可区分。这个实验是第一个证明牡蛎幼虫摄入微球以外的塑料颗粒的实验。本方法有助于在实验室实验中使用小微纤维,从而更全面地了解环境中的微塑料效应。环境毒物化学2022;41:944–953。© 2021 SETAC
更新日期:2021-09-22
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