Preparation of Xenopus laevis retinal cryosections for electron microscopy

• A method for preparing X. laevis retina samples for electron microscopy.
• The method involves an intermediate cryosectioning step.
• Reproducible orientation and improved throughput with excellent preservation.
• The method also works well using samples fixed for fluorescence microscopy.
• The method is amenable to dual light/electron microscopy paradigms.

Transmission electron microscopy is the gold standard for examination of photoreceptor outer segment morphology and photoreceptor outer segment abnormalities in transgenic animal models of retinal disease. Small vertebrates such as zebrafish and Xenopus laevis tadpoles have been used to generate retinal disease models and to study outer segment processes such as protein trafficking, and their breeding capabilities facilitate experiments involving large numbers of animals and conditions. However, electron microscopy processing and analysis of these very small eyes can be challenging. Here we present a methodology that facilitates processing of X. laevis tadpole eyes for electron microscopy by introducing an intermediate cryosectioning step. This method reproducibly provides a well-oriented tissue block that can be sectioned with minimal effort by a non-expert, and also allows retroactive analysis of samples collected on slides for light microscopy.