A practical protocol to prepare paraffin-embedded whole tick histology sections.
A practical protocol to prepare paraffin-embedded whole tick histology sections.
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DOI：
10.1016/j.ttbdis.2023.102162
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发表时间：
2023-07
影响因子：
3.2
通讯作者：
Birkenheuer AJ
中科院分区：
文献类型：
--
作者：
Yang TS;LaDouceur EEB;Baumgartner WA;Marr HS;Karounos M;Robertson J;Whitehurst N;Miller LS;Birkenheuer AJ
蜱是重要的体外寄生虫，能够传播多种类型的病原体，目前与世界范围内许多新兴的蜱传疾病有关。随着人类和兽医物种中蜱传疾病的发生率越来越高，我们不断需要进一步了解蜱及其传播的病原体。全蜱组织学提供了蜱虫内部解剖的完整范围，使研究人员能够在一个切片中检查多个感兴趣的器官。这与蜱传疾病研究中更常用的其他技术相反，例如电子显微镜和单个器官的光学显微镜。目前缺乏文献描述一种实用的技术来处理整个蜱组织切片。因此，目前的研究旨在为研究人员提供一个可行的方案，以制备高质量的石蜡包埋的全蜱组织学切片。Amblyomma americanum成虫用作本研究的示例物种。在使用各种固定剂、软化剂和加工技术的组合进行了一系列中试实验后，我们选择比较两种常见的固定剂，10%中性缓冲福尔马林（NBF）和布因溶液用于全蜱。数量相等的A。对未进食的美洲蜱成虫（n = 10/固定剂）进行相同的处理，并对它们的整个蜱组织学冠状切片进行单独评分。包含更多对蜱传疾病研究至关重要的内部器官（例如唾液腺和中肠）、切片上组织和外骨骼的高度完整性以及组织的良好固定和染色质量的完整蜱切片得分较高。布因固定蜱的平均总评分显著高于NBF固定蜱（p = 0.001）。为了进一步评估我们的首选技术，我们还证明了使用布因溶液生产其他三种具有医学重要性的常见蜱种（血红扇头蜱、肩胛硬蜱和变异革蜱）的高质量全蜱切片的可行性。虽然这种技术可能需要进一步优化其他蜱种属，但我们描述了一种使用常用工具、试剂和标准组织学设备的可行方案。这应该允许任何研究者根据他们的实验目标根据需要容易地对该方案进行调整。
Ticks are important ectoparasites that are capable of transmitting multiple classes of pathogens and are currently linked with many emerging tick-borne diseases worldwide. With increasing occurrences of tick-borne diseases in both humans and veterinary species, there is a continuous need to further our understanding of ticks and the pathogens they transmit. Whole tick histology provides a full scope of the tick internal anatomy, allowing researchers to examine multiple organs of interest in a single section. This is in contrast to other techniques that are more commonly utilized in tick-borne disease research, such as electron microscopy and light microscopy of individual organs. There is a lack of literature describing a practical technique to process whole tick histologic sections. Therefore, the current study aims to provide researchers with a workable protocol to prepare high quality paraffin-embedded whole tick histology sections. Amblyomma americanum adults were used as an example species for this study. After a series of pilot experiments using a combination of various fixatives, softening agents and processing techniques, we elected to compare two common fixatives, 10% neutral-buffered formalin (NBF) and Bouin’s solution for whole ticks. Equal numbers of A. americanum unfed adults (n = 10/fixative) were processed identically and their whole tick histology coronal sections were individually scored. Higher scores were assigned to whole tick sections that contained more internal organs that are crucial for tick-borne disease research (e.g. salivary glands and midgut), high integrity of tissues and exoskeleton on the section, and good fixation and staining quality of the tissues. The mean total scores for Bouin’s-fixed ticks were significantly higher compared to NBF-fixed ticks (p = 0.001). To further assess our preferred technique, we also demonstrated the feasibility of producing high quality whole tick sections for three other common tick species of medical importance (Rhipicephalus sanguineus, Ixodes scapularis, and Dermacentor variabilis) using Bouin’s solution. While this technique may require further optimization for other tick species, we described a feasible protocol that uses commonly available tools, reagents and standard histologic equipment. This should allow any investigator to easily make adjustments to this protocol as needed based on their experimental goals.
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影响因子：
5.7
作者：
Hermance, Meghan E.;Thangamani, Saravanan
通讯作者：
Thangamani, Saravanan
DOI：
10.1111/j.1550-7408.1977.tb00974.x
发表时间：
1977-01-01
期刊：
JOURNAL OF PROTOZOOLOGY
影响因子：
--
作者：
MEHLHORN, H;SCHEIN, E
通讯作者：
SCHEIN, E
影响因子：
1.9
作者：
Mihara, Ryo;Umemiya-Shirafuji, Rika;Suzuki, Hiroshi
通讯作者：
Suzuki, Hiroshi
影响因子：
2.6
作者：
Saito, KC;Bechara, GH;Mathias, MIC
通讯作者：
Mathias, MIC
影响因子：
2.7
作者：
Starck, J. Matthias;Mehnert, Lisa;Hoernig, Marie
通讯作者：
Hoernig, Marie