Simple Way To Measure Ethanol Sensitivity In Flies l Protocol Preview

Summary

In this video, Thomas Maples from UT Southwestern Medical Center demonstrates a simple procedure to test the sensitivity of fruit flies to ethanol-induced sedation. The assay is based on the flies' loss of righting reflex and walking ability after ethanol exposure, which can reveal differences in alcohol sensitivity among various Drosophila strains and mutants. The method also allows for inducing alcohol tolerance in flies. The experiment requires flies, an empty food vial, a razor, cotton-like plugs, a micro pipette, 200 proof ethanol with food dye, and a timer.

Takeaways

• 🔬 Thomas Maples is demonstrating an assay to determine the sensitivity of fruit flies to ethanol-induced sedation.
• 🧪 The assay is based on the loss of righting reflex and walking ability in flies after ethanol exposure.
• 📊 This method can be used to compare alcohol sensitivity between different strains of Drosophila and mutants.
• 🔄 The assay can also be used to induce tolerance in flies, which is a reduced response to alcohol after repeated exposure.
• 🧳 The experiment requires vials with eight fruit flies, an empty food vial, a razor, cotton-like vial plugs, a micro pipette, 200 proof ethanol with food dye, and a timer.
• 🐛 Males or females can be used for the exposures, but each vial should contain only one sex.
• 🔪 To create an exposure chamber, a vial plug is cut in half using a razor blade.
• 🔎 The flies' mobility will be monitored in the exposure chamber to assess their sensitivity to ethanol.
• 📝 The procedure involves exposing the flies to ethanol and observing their behavior to measure alcohol sensitivity.
• 🔴 A 200 proof ethanol solution with food dye is used for the exposure, which is easily obtainable at a grocery store.

Q & A

Who is Thomas Maples and what is his role in the video?

Thomas Maples is from the lab of Dr. Adrian Rothenflu in the Department of Psychiatry at UT Southwestern Medical Center in Dallas. He is demonstrating a procedure in the video.

What is the main purpose of the procedure demonstrated by Thomas Maples?

The procedure aims to determine the sensitivity of flies to ethanol-induced sedation, which is based on the flies' loss of righting reflex and walking ability after exposure to ethanol.

What can the assay measure using this procedure?

The assay can measure alcohol sensitivity differences between different strains of Drosophila and mutants, as well as induce tolerance in flies, which is a reduced response to alcohol after repeated exposure.

What materials are needed for this experiment?

The materials needed include vials with eight fruit flies each, an empty food vial, a razor, two vial plugs made of cotton-like material, a 1000 microliter micropipette with tips, 200 proof ethanol with food dye, and a timer.

Why is it important to use flies that are one or two days old for the experiment?

Flies that are one or two days old are used because they are likely more uniform in age and physiological state, which can help ensure consistency in the experiment's results.

Why should each vial contain only one sex of flies?

Each vial should contain only one sex of flies to control for any potential differences in sensitivity to ethanol between males and females, ensuring the results are not confounded by sex-related variables.

What is the role of the cotton-like material or cotton balls in the experiment?

The cotton-like material or cotton balls are used to make vial plugs, which are necessary for creating an airtight seal when exposing the flies to ethanol.

How is the exposure chamber created for the flies?

The exposure chamber is created by cutting a vial plug in half using a razor blade and pushing one of the halves to the bottom of the vial.

What is the significance of using 200 proof ethanol in the experiment?

200 proof ethanol is used because it is a high concentration of ethanol that effectively induces sedation in the flies, allowing for a clear measurement of their sensitivity.

Why is food dye added to the ethanol?

Food dye is added to the ethanol to visually confirm that the ethanol is being properly aerosolized and distributed within the exposure chamber.

How does the timer play a role in the experiment?

The timer is used to standardize the exposure time of the flies to ethanol, ensuring that all flies are subjected to the same conditions for a consistent and controlled experiment.