Our experiments are performed after blinding ie we do not know the genotype of the flies tested. Code for this line is Red2. Intensity was approximately 400 Lux. The frequency of yellow light was 20Hz with 50:50 duty cycle.

Our experiments are performed after blinding ie we do not know the genotype of the flies tested. Code for this line is Orange3. Intensity was approximately 400 Lux. The frequency of yellow light was 20Hz with 50:50 duty cycle.

Yellow 1 (Positive Control):  Gr28bd-G4, TrpA1-G4

Light: intensity (500 Lux side, 1000 Lux bottom), frequency (20Hz)

The platform experiment was done with us being blinded. We referred to the genotype as the Red1 Genotype. The light intensity was taken as approximately 400 lux as was decided by previous experiments. The duty cycle was 50:50 at a frequency of 20 Hz.

Used positive control flies (NorpA; UAS Crimson) to check what intensity of light is better for activating the crimson channel and for studying learning behaviour in the joystick experiment.  Tested light in 3 intensity ranges to check what would be best suited.

1. Initially, we were doing experiments at 700 lux intensity. We saw results which possibly point out to damaging of neuronal populations due to such high-intensity light. As can be seen in the graph below, there is a strong aversion in the first 2 minutes of the training period. However, this aversion drastically decreases later and a very strong phenotype is not shown later. This may prove the hypothesis that high-intensity light(about 700 lux) may result in damage to neuronal populations.
2. So we tried with two more light intensities – 400 lux and 100 lux. The graphs for these are attached below. As can be clearly seen that now the results dont just show up for the first training segment but remain til the very end. This indicates that both 100lux and 400 lux light intensity are safe and do not disrupt neurons. Also, it can be clearly seen that there is a stronger and more pronounced effect with 400 lux light intensity. Thus we decide that 400 lux light intensity needs to be used for testing crimson channel expression for the joystick experiment.

Blue 1 combined PI value plot at 400 lux at 50 per cent duty cycle and 20 Hz

Trajectories of blue 1 flies on drosophila joystick apparatus shown:-

Comparison between White 1 ( Control (NorpA- UAS Chrimson)) and Yellow 1 (Positive Control ( Gr28bd-G4, TrpA1-G4).

To test whether the flies are really blind, and there is no problem with the NorpA part of the construct, we compare with NorpA,UAS GTACR1 ; NorpA,UAS GTACR2 ; and another stock of NorpA-UAS-Chrimson.

Weighted mean is calculated by multiplying the weights (total number of flies in that experiment/total number of flies in all the experiments) with the PI for that experiment, and taking the sum thereafter for all the experiments.

the first graph shows Bar plot of Mean and standard deviation of PIs for Genetic control (n=9) and Positive control (n=5).
The second graph shows weighted mean and weighted standard deviation of the same.

Weighted mean is calculated by multiplying the weights (total number of flies in that experiment/total number of flies in all the experiments) with the PI for that experiment, and taking the sum thereafter for all the experiments.

Positive control not working: why?