Drosophila Burrowing and Tunneling Assay: A Method to Assess Tissue Hypoxia in Fly Larvae

Abstract

Source: Qiang, K. M., et al. A Burrowing/Tunneling Assay for Detection of Hypoxia in Drosophila melanogaster Larvae. J. Vis. Exp. (2018).

This video shows a method to test hypoxia in Drosophila larvae by assessing their burrowing and tunneling abilities. Using this technique researchers can distinguish larvae experiencing oxygen deprivation from those with developmental deficits. The example protocol demonstrates the setup for this assay.

Protocol

This protocol is an excerpt from Qiang et al., A Burrowing/Tunneling Assay for Detection of Hypoxia in Drosophila melanogaster Larvae, J. Vis. Exp. (2018). 1. Preparation of larvae Two or more days before beginning the assay, set up crosses of the desired experimental and control combinations (minimum 20 females and 10 males each) in egg-lay dishes as described by Wieschaus and Nusslein-Volhard but using 50 mL polypropylene beak…

Materials

REAGENTS
Dehydrated yeast
Frozen grape juice concentrate	Welch's		Available at most large supermarkets
Glacial acetic acid	Sigma-Aldrich	320099
Drosophila agar	Apex Bioresearch Products	66-103
Methyl-para-hydroxybenzoate	Apex Bioresearch Products	20-658
EQUIPMENT
50 ml polypropylene beakers
6.0 cm disposable Petri dishes	Falcon	08757100B
10 cm disposable plastic Petri dishes	E+K Scientific	EK-24104
Plastic microspatulas	Corning Incorporated	3012
Bent teasing needle	Nasco	S08848MH
Dissecting microscope			Any microscope with 10-30X magnification