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

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

1. For the Joystick experiment – We continuously varied the light intensity at the tip of the optical fibre and measured it using the Lux meter at first and then in mW/cm^2. Then both these readings were taken down and the best fit was generated which depicted the measured fit. Now a theoretical fit was also generated and plotted. This is given below
2. We also did the best fit for the Tmaze experimental setup. We measured the light intensity on both sides of the chamber in between and took the average of it. 
3. The Information for plotting the theoretical fit was obtained from here: https://www.dfisica.ubi.pt/~hgil/Fotometria/HandBook/ch07.html

   ## 1 Watt = 517.031 lm at 590 nm

   ## 1 lux = 1 lm/m2

   ## Therefore: 517.031 lux = 517.031 lm/m2 = 1 Watt/m2 = 1000mW/mm2 = 10mW/cm2

   Light Measurement Handbook: Measurement Geometries

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.

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.

After getting a new spectrometer, we confirmed that the first one was faulty. Comparison of old (1st and 3rd measures) and new (2nd and 4th) spectrometer. Now much more sensitive and the right spectrum measured for the green LED present in the spectrometer itself and for the light comming out of the light guide coupled with the red LED (whose spectrum does not seem to change after travelling through the light guide)

This is the first line I completed from the screen. What it is more important is that I found out that NorpA;UAS-Chrimson has orange eyes, when it should be red. So I think I will start to do the line again and meanwhile test the contaminated line to see what the phenotype looks like

We need a student to finish up experiments

Example of ON/OFF traces.  And then the wiggling was measured in both conditions (with and without light). It seems like they move more when they are in the light side. I also did measure the derivative with a tau=2 (in case the sampling is over the temporal frequency of fly behavior)

Comparing scores in both setups show some lines that match their scores and other lines that have opposite scores. This filters the ones that are context dependent to the ones that show a context independent reinforcement valence.