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Immunofluorescent staining and imaging of the pupal and adult Drosophila visual system

Nature Protocols volume 1, pages 2635–2642 (2006)Cite this article

Abstract

This immunofluorescence protocol can be used to assay cell morphology, cell positioning and subcellular localization of proteins in the fly eye at stages of development from early pupation to adult. The protocol includes the following procedures: collecting and developmentally staging Drosophila pupae, dissecting fly eyes at defined stages of development, immunostaining of retina and preparing visual system samples (i.e., retina and optic lobe) for confocal microscopy. It is supplemented with images of key dissection steps, guidelines for troubleshooting and examples of data obtained using these methods. Overall, this protocol takes up to 9 d to complete. The amount of hands-on time required on each day varies, ranging from 30 min to several hours depending on the number of stages and/or genotypes one wishes to study.

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Figure 1: Dissecting station.
Figure 2: Pupal visual system dissection.
Figure 3: Immuno-imaging of whole mount retinas and optic lobes

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Acknowledgements

The authors would like to thank Donald Ready (Purdue University), who has pioneered the development of the protocols and the operating mode described in this manuscript. We would also like to thank the Pichaud lab for comments and suggestions about the manuscript. R.F.W. is the recipient of a National Cancer Institute of Canada Post-PhD Research Fellowship. This work is funded by an MRC Career Track Award to F.P.

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  1. Department of Anatomy and Developmental Biology, MRC Laboratory for Molecular Cell Biology and Cell Biology Unit, University College London, Gower Street, London, WC1E 6BT, UK

    Rhian F Walther & Franck Pichaud

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  1. Rhian F Walther
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  2. Franck Pichaud
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Correspondence to Franck Pichaud.

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Walther, R., Pichaud, F. Immunofluorescent staining and imaging of the pupal and adult Drosophila visual system. Nat Protoc 1, 2635–2642 (2006). https://doi.org/10.1038/nprot.2006.379

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