Techniques | fly muscles lab

Handling fruit flies

At the core of our scientific work is the fundamental and rewarding task of placing flies in a tube and waiting for them to reproduce. Many genetically complex experiments can be conducted in Drosophila, such as knocking down genes, creating mosaic animals, and expressing fluorescently tagged proteins. Most of these experiments involve crossing flies, so we do that quite frequently.

Arduino-based and 3D-printed tools

Sometimes, we need to create our own tools to handle flies in specific ways. This is where Arduino microcontrollers and 3D printers come to the rescue. Two tools we’ve recently developed are:

An LED infrared device to track the movement of individual fly wings. We used this to test whether wings can become uncoordinated in certain mutant lines—and indeed, they can.
A flyflipper machine that flips the orientation of tubes, encouraging flies to walk for extended periods of time.

Confocal Microscopy.

We have a Thorlabs 4-channel confocal microscope in the lab. We also have a good collection of Drosophila strains with muscle proteins tagged with GFP or mCherry. So we dissect the muscles and see them under the microscope. We use the microscope to characterize the muscle defects in different mutant and RNAi conditions.

Transmission electron microscopy

We can access electron microscopy thanks to the Electron Microscopy Core Facility and Mary-Ann Trevors. We section the leg of the indirect flight muscles of different genetic backgrounds of Drosophila.

In-vivo protein biotinylation

We use proteomic approaches as an unbiased approach to find Z-disc proteins in different genetic conditions and to find the proteins bound to filamins in muscles. Both approaches rely on the biotin ligase called TurboID. A bait protein fused to a biotin ligase catalyzes the biotinylation of neighboring proteins within specific cells. We have a Z-disc TurboID variant that allows sus to interrogate the Z-disc proteome in different conditions. Then, using nanobodies, we are testing the filamin proteome in different mutants

AlphaFold and protein modelling

AlphaFold and similar programs allow us to guess the structure of proteins based on their amino acid sequence. We use them to predict the binding sites of filamins. It's a great tool because it allows us to test many proteins relatively quickly. We are also looking into ways of testing the predicted interactions in vivo.

Sometimes, we need to create our own tools to handle flies in specific ways. This is where Arduino microcontrollers and 3D printers come to the rescue. Two tools we’ve recently developed are:

An LED infrared device to track the movement of individual fly wings. We used this to test whether wings can become uncoordinated in certain mutant lines—and indeed, they can.

A flyflipper machine that flips the orientation of tubes, encouraging flies to walk for extended periods of time.