Comparative analysis of fixation and embedding techniques for optimized histological preparation of zebrafish.
Comparative analysis of fixation and embedding techniques for optimized histological preparation of zebrafish.
复制标题
DOI：
10.1016/j.cbpc.2017.11.003
复制
发表时间：
2018-06
期刊：
影响因子：
--
通讯作者：
Cheng KC
中科院分区：
文献类型：
--
作者：
Copper JE;Budgeon LR;Foutz CA;van Rossum DB;Vanselow DJ;Hubley MJ;Clark DP;Mandrell DT;Cheng KC
认识到斑马鱼作为研究人类疾病的模式生物的重要性，我们为基于网络的显微解剖学参考图谱创建了斑马鱼内容，用于比较模型系统和人类之间的组织学和组织病理学（http：bio-atlas.psu.edu）。固定，脱钙，包埋和切片的斑马鱼进行了优化，以最大限度地提高切片质量。涉及六种固定剂的方案的比较表明，10%中性缓冲福尔马林在21 °C下24小时产生优异的结果。幼鱼和成年鱼的切片需要进行骨脱钙; 0.35 M EDTA在21日龄幼鱼至成年鱼（≥ 3个月）中产生有效脱钙。为了提高切片平面的一致性，在套幼虫，我们已经开发了新的阵列铸造模具的基础上，来自3D microCT图像的幼虫的外部轮廓。组织不连续的部分，一个共同的障碍，以创建高质量的部分斑马鱼，最大限度地减少处理和嵌入的福尔马林固定的斑马鱼组织在塑化形式的石蜡，并定期水合块表面在冰水之间的部分。最佳H&E（苏木精和伊红）染色是通过改进标准方案实现的。由玻璃组织学切片生成的高质量切片扫描经过数字处理，以最大限度地提高图像质量，并将实验复制品作为完整切片发布，作为本出版物的一部分。仍然需要对组织处理进行修改，以消除对块表面水合的需要。进一步增加来自其他模型系统的幻灯片集合和用于可视化组织结构的3D工具将大大增加数字图谱的实用性。
In recognition of the importance of zebrafish as a model organism for studying human disease, we have created zebrafish content for a web-based reference atlas of microanatomy for comparing histology and histopathology between model systems and with humans (http://bio-atlas.psu.edu). Fixation, decalcification, embedding, and sectioning of zebrafish were optimized to maximize section quality. A comparison of protocols involving six fixatives showed that 10% Neutral Buffered Formalin at 21 °C for 24 h yielded excellent results. Sectioning of juveniles and adults requires bone decalcification; EDTA at 0.35 M produced effective decalcification in 21-day-old juveniles through adults (≥~3 Months). To improve section plane consistency in sets of larvae, we have developed new array casting molds based on the outside contours of larvae derived from 3D microCT images. Tissue discontinuity in sections, a common barrier to creating quality sections of zebrafish, was minimized by processing and embedding the formalin-fixed zebrafish tissues in plasticized forms of paraffin wax, and by periodic hydration of the block surface in ice water between sets of sections. Optimal H&E (Hematoxylin and Eosin) staining was achieved through refinement of standard protocols. High quality slide scans produced from glass histology slides were digitally processed to maximize image quality, and experimental replicates posted as full slides as part of this publication. Modifications to tissue processing are still needed to eliminate the need for block surface hydration. The further addition of slide collections from other model systems and 3D tools for visualizing tissue architecture would greatly increase the utility of the digital atlas.
登录
查看更多内容
影响因子：
2.7
作者：
Moore, JL;Aros, M;Cheng, KC
通讯作者：
Cheng, KC
影响因子：
2.5
作者：
KIMMEL, CB;BALLARD, WW;SCHILLING, TF
通讯作者：
SCHILLING, TF
影响因子：
5
作者：
Beckwith, LG;Moore, JL;Cheng, KC
通讯作者：
Cheng, KC
影响因子：
2.7
作者：
Tsao-Wu, GS;Weber, CH;Cheng, KC
通讯作者：
Cheng, KC
DOI：
10.1186/s13046-015-0196-8
发表时间：
2015-08-12
期刊：
Journal of experimental & clinical cancer research : CR
影响因子：
--
作者：
Zhao S;Huang J;Ye J
通讯作者：
Ye J