Ramirez-Esquivel Fiorella, Ribi Willi A, Narendra Ajay
Research School of Biology, Australian National University.
Department of Biological Sciences, Macquarie University;
J Vis Exp. 2017 Nov 27(129):56339. doi: 10.3791/56339.
This article outlines a suite of techniques in light microscopy (LM) and electron microscopy (EM) which can be used to study the internal and external eye anatomy of insects. These include traditional histological techniques optimized for work on ant eyes and adapted to work in concert with other techniques such as transmission electron microscopy (TEM) and scanning electron microscopy (SEM). These techniques, although vastly useful, can be difficult for the novice microscopist, so great emphasis has been placed in this article on troubleshooting and optimization for different specimens. We provide information on imaging techniques for the entire specimen (photo-microscopy and SEM) and discuss their advantages and disadvantages. We highlight the technique used in determining lens diameters for the entire eye and discuss new techniques for improvement. Lastly, we discuss techniques involved in preparing samples for LM and TEM, sectioning, staining, and imaging these samples. We discuss the hurdles that one might come across when preparing samples and how best to navigate around them.
本文概述了一套光学显微镜(LM)和电子显微镜(EM)技术,可用于研究昆虫的眼内外解剖结构。这些技术包括针对蚂蚁眼睛研究进行优化并与其他技术(如透射电子显微镜(TEM)和扫描电子显微镜(SEM))协同使用的传统组织学技术。这些技术虽然非常有用,但对于新手显微镜工作者来说可能具有挑战性,因此本文着重介绍了针对不同标本的故障排除和优化方法。我们提供了关于整个标本成像技术(光学显微镜和扫描电子显微镜)的信息,并讨论了它们的优缺点。我们重点介绍了用于确定整个眼睛晶状体直径的技术,并讨论了改进的新技术。最后,我们讨论了用于光学显微镜和透射电子显微镜的样品制备、切片、染色和成像技术。我们讨论了在制备样品时可能遇到的障碍以及如何最好地克服这些障碍。