Ultrastructural Analysis of Drosophila Ovaries by Electron Microscopy

Abstract

i. Summary The Drosophila melanogaster ovary is a powerful, genetically tractable system through which one can elucidate the principles underlying cellular function and organogenesis in vivo. In order to understand the intricate process of oogenesis at the subcellular level, microscopic analysis with the highest possible resolution is required. In this chapter, we describe the preparation of ovaries for ultrastructural analysis using transmission electron microscopy and focused ion beam scanning electron microscopy. We discuss and provide protocols for chemical fixation of Drosophila ovaries that facilitate optimal imaging with particular attention paid to preserving and resolving mitochondrial membrane morphology and structure.

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“…Transmission electronic microscopy (TEM). TEM was performed similar to a previous study 95 , with minor modifications. Fly ovaries were immersed in fixation solution (2.5% glutaraldehyde, 1% tannic acid and 2% paraformaldehyde in 0.1 M sodium cacodylate buffer) overnight at 4 °C.…”