on Tuesday, June 19th, 2018 12:13 | by Ottavia Palazzo
on Monday, June 18th, 2018 1:15 | by Gaia Bianchini
Yellow 1 (Positive Control): Gr28bd-G4, TrpA1-G4
Light: intensity (500 Lux side, 1000 Lux bottom), frequency (20Hz)
on Friday, June 8th, 2018 11:47 | by Ottavia Palazzo
Not confocal immages (going to the confocal this week)
The construct I am analyzing is the FoxP-IsoB_Gal4, so with the Gal4 inserted in the 8th exon
on Monday, January 29th, 2018 12:40 | by Christian Rohrsen
Some traces from this week just so that you have an idea how do they look like. To me they are not the optimal traces I expected. But one can see some signal there. I will start the screen hoping to get enough good traces without too much work.
what do you think is the best quality control for accepting a trace for the analysis or not. I was thinking the 3D mapping gives a good hint but without quantification.
on Wednesday, February 13th, 2013 11:40 | by Christine Damrau
As a pilot for future experiments I tried to reproduce results from Keene et al., 2012. There, they substituted sugar by a laser to provoke PER after 24h of starvation (females, one week old). In TH-GAL4 they found 100% response when they pointed the laser to head or thorax. In tdc2-GAL4 indeed they found 50% of flies responding to heat but only when the laser was pointed to the thorax but not to the head.
Since we do not have this kind of laser, they used, I played around with an infrared light. Flies were fixed to hooks and a clamp as always. First, there was given a filter paper soaked with EVIAN-water (negative control), then there were 10s of heat (36-38°C), afterwards two positive controls: 30% sucrose on the legs and finally on the labellum.
In the figure, you can see that I am able to reproduce the methods with an infrared lamp instead of a laser. The TH-GAL4 flies crossed to TrpA respond much more to heat compared to the controls (see figure in blue and red). The proboscis extension appears already after a few seconds.
Unfortunately, I could not reproduce the results for tdc2-GAL4 (see figure in green). There was no response to heat found at all. (tdc2-GAL4 has to be ok, because I see fluorescence when crossed to GCaMP). Interestingly, the flies did not respond as strong to the 30% sucrose as expected. The starvation may have to be longer to increase motivation?
Next step will be to see a phenotype that can be seen by activating tdc2-GAL4. That may be PER after longer starvation or flight behavior.
on Tuesday, December 4th, 2012 12:49 | by Julien Colomb
Since we have problems with Madeleine’s project, I will make crosses to test that the flies are ok before using them for PKC RNAi experiments.
the putative elavGal$;tubGal80ts flies will be crossed to w; AKAP/Cyo flies. Then the son ElavGal4/Y, tubGal80ts/+ will be crossed to w.
Only non Cyo male flies should get white eyes, if not, the stock is not clean.
In parallel, I will also start crossing nsybGal4 into the tubGal80ts line (2 to 4 crosses necessary depending on the eye color of the heterozygotes).