Mitchell Dorothy G, Edgar Allison, Martindale Mark Q
The Whitney Laboratory for Marine Bioscience, 9505 N, Ocean Shore Blvd, St. Augustine, FL, 32080-8610, USA.
Front Zool. 2021 Jun 12;18(1):29. doi: 10.1186/s12983-021-00414-z.
Gelatinous zooplankton can be difficult to preserve morphologically due to unique physical properties of their cellular and acellular components. The relatively large volume of mesoglea leads to distortion of the delicate morphology and poor sample integrity in specimens prepared with standard aldehyde or alcohol fixation techniques. Similar challenges have made it difficult to extend standard laboratory methods such as in situ hybridization to larger juvenile ctenophores, hampering studies of late development.
We have found that a household water repellant glass treatment product commonly used in laboratories, Rain-X®, alone or in combination with standard aldehyde fixatives, greatly improves morphological preservation of such delicate samples. We present detailed methods for preservation of ctenophores of diverse sizes compatible with long-term storage or detection and localization of target molecules such as with immunohistochemistry and in situ hybridization and show that this fixation might be broadly useful for preservation of other delicate marine specimens.
This new method will enable superior preservation of morphology in gelatinous specimens for a variety of downstream goals. Extending this method may improve the morphological fidelity and durability of museum and laboratory specimens for other delicate sample types.
由于其细胞和非细胞成分独特的物理特性,凝胶状浮游动物在形态学上可能难以保存。相对较大体积的中胶层会导致使用标准醛类或酒精固定技术制备的标本中,其精致的形态发生扭曲,样本完整性较差。类似的挑战使得将原位杂交等标准实验室方法应用于较大的幼年栉水母变得困难,阻碍了后期发育的研究。
我们发现,实验室常用的一种家用防水玻璃处理产品Rain-X®单独使用或与标准醛类固定剂结合使用时,能大大改善此类精致样本的形态保存。我们给出了与长期保存或检测和定位目标分子(如免疫组织化学和原位杂交)兼容的不同大小栉水母保存的详细方法,并表明这种固定方法可能广泛适用于保存其他精致的海洋标本。
这种新方法将能够为各种下游目标在凝胶状标本中实现卓越的形态保存。推广这种方法可能会提高博物馆和实验室中其他精致样本类型标本的形态保真度和耐久性。