Name: a novel rfp reporter to aid in the visualization of the eye imaginal disc in drosophila
Uploaded: 2009-12-15T17:00:00
Description: Watch this Scientific Journal Video about A Novel RFP Reporter to Aid in the Visualization of the Eye Imaginal Disc in Drosophila at JoVE.com

This work was funded by King&#x2019;s College London and the Royal Society.

<ol>
 <li>First place a drop of PBS onto a Sylgard dish under a dissecting microscope. </li>
 <li>Next remove several wandering third instar larvae from a vial or bottle using a spatula and place them in the PBS.</li>
 <li>To begin the dissection use your left hand tweezers (if you are right handed) to grasp a larva two thirds of the way towards the posterior and then use the other pair to hold the dark colored mouth hooks at the anterior end. </li>
 <li>Now gently pull the larva apart by moving both pairs of tweezers away fro...

<ol>
	<li>Doroquez, D. B., Rebay, I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Crit Rev Biochem Mol Biol. 41 (6), 339-339 (2006).</li><li>Kaul, A. K., Craig, G. M., McNeill, H., Bateman, J. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> , .</li><li>Barolo, S., Castro, B., Posakony, J. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Biotechniques. 36 (3), 436-436 (2004).</li></ol>

Dissection of the Drosophila eye imaginal disc is straightforward, but can take time to master. The critical steps are the identification of the eye disc and its dissection away from the larval brain. Identification of the eye disc is made easier using the eye-RFP line described here. Possible modifications are to remove the antennal disc or the peripodial membrane, or to leave the optic lobe attached to visualize the PR axons entering the brain.
The eye-RFP line was generated by clo...

a novel rfp reporter to aid in the visualization of the eye imaginal disc in drosophila

The Drosophila eye is a powerful model system for studying areas such as neurogenesis, signal transduction and neurodegeneration. Many of the discoveries made using this system have taken advantage of the spatiotemporal nature of photoreceptor differentiation in the developing eye imaginal disc. To use this system it is first necessary for the researcher to learn to identify and dissect the eye disc. We describe a novel RFP reporter to aid in the identification of the eye disc and the visualization of specific cell types in the developing eye. We detail a methodology for dissection of the eye imaginal disc from third instar larvae and describe how the eye-RFP reporter can aid in this dissection. This eye-RFP reporter is only expressed in the eye and can be visualized using fluorescence microscopy either in live tissue or after fixation without the need for signal amplification. We also show how this reporter can be used to identify specific cells types within the eye disc. This protocol and the use of the eye-RFP reporter will aid researchers using the Drosophila eye to address fundamentally important biological questions.

Watch this Scientific Journal Video about A Novel RFP Reporter to Aid in the Visualization of the Eye Imaginal Disc in Drosophila at JoVE.com

A Novel RFP Reporter to Aid in the Visualization of the Eye Imaginal Disc in Drosophila

We describe a novel red fluorescent protein (RFP) reporter that is expressed specifically in the Drosophila eye. We detail a methodology for dissection of the eye imaginal disc and how this reporter can be used to aid in the dissection and identification of specific cell types in the developing eye.

A Novel RFP Reporter to Aid in the Visualization of the Eye Imaginal Disc in Drosophila

The Drosophila eye is a powerful model system for studying areas such as neurogenesis, signal transduction and neurodegeneration. Many of the discoveries ...

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