Fly’s Neuronal Activity Experiment Setup using Two-Photon Microscopy

Supervisor: Professor Jean-Michel Mongeau

As a graduate student in the laboratory, I have participated in research focused on recording the activities of neurons in flies in order to understand the connection between neuron control and bio-motion. I was trained to tether fruit flies to the fly holder without causing any injury. Taking on a leading role in the experimental setup, I constructed the fly holder according to the previous design and created a mounting system with adjustable angles for the semicylinder LED arena. Additionally, I developed a temperature control saline system for the fly holder in the two-photon microscopy area. Furthermore, I received training in aligning the Sapphire Femtosecond Laser for Two-Photon Microscopy.

During this research, I carried out the following tasks:

• Learned to tether fruit flies to the static and flying fly holders.
• Learned and documented the fruit flies’ head dissection to explore the brain under the 2-photon microscopy.
• Constructed the flying fly holders and the brass tube holder in the anesthesia system in the learning factory.
• Designed and developed a versatile angle-adjustable mounting system for the arena and fly holder. Constructed the bill of material (BOM) and successfully assembled all the components.
• Designed and developed a temperature control saline system for the fly holder. Constructed the BOM and successfully assembled all the components.
• Received training in aligning the laser.
• Crossed flies in the GAL4-UAS system to obtain the GAL4-GCaMP flies for 2-photon microscopy.
• Imaged the calcium activity in neurons of flies in 2-photon microscopy.

(a) The 2-photon experiment setup in SolidWorks. (b) The real setup. Changed the fly holder mounting stage to the left side for a better user experience when finding the head under microscopy. (c) Saline system for the fly holder to maintain the temperature of the brain. (d) Calcium imaging of the fly's brain.

Tethered fly is flying