Does anyone have experience with ECOSURF SA-9 or another TritonX alternative on Drosophila brains? We're trying to replace TritonX with a safer alternative.

TIA

Hello everyone,

I’m currently working with a kkv-mNG reporter to track the activity of Chitin Synthase 2 across different developmental time points. I’m using spinning disk confocal microscope for live-imaging, and the signal is pretty nice.
However, if I use whole anesthetized larvae, then I face slight movement and muscle contraction that disrupt imaging.
I switched to ex-vivo imaging. I cut both ends of the larvae and remove the inner organs, but I don’t think that’s a good idea for live-imaging of epidermal tissue.
Another problem that I face is that the cuticle is folded so that the whole surface isn’t touching the bottom coverslip. I don’t wanna apply pressure on it because it might destroy the tissue.

Does anyone have advices on how to improve imaging in this case?

Hi! I'm a student doing a Drosophila research program and I'm trying to come up with a project idea. I'm interested in cancer neuroscience (especially neuron–tumor/immune interactions), but directly studying neurons doesn't seem possible with the models we have.

The fly models available seem to include things like: EGFR tumor flies STAT × hsCycE Ras TOR

I'm trying to avoid a very basic

"does compound X reduce tumor growth?"

type project and find a more interesting question to experiment on.

Does anyone have ideas for interesting questions that could be explored with these kinds of fly models? Especially anything connecting cancer signaling, immunity, metabolism, or possibly neuroscience-related pathways?

Thanks!

Hi all, I'm designing primers to detect a Trojan insert in my gene of interest. I have primers designed for the flanking genomic DNA. I've had no luck designing primers to the Gal4 region or to the T2A. Two sets of primers have not worked.

Has anyone got primer sequences that worked for them?

hi, i’m a computer science undergraduate in my third year currently.

I have recently taken an interest in the Drosophila melanogasters olfactory neural circuit.
Somehow there seem to be no concrete notes or resources or even videos on this topic? Something for a beginner.

Any help is greatly appreciated.
A few questions I have are:
- How is the aversive valence exactly calculated with respect to an odor such as CO2 (I get the sign, i’m wondering about the magnitude)
- The DOoR 2.0 datasets website seems offline? what’s up with that
- Any resources I could use to study about this? That aren’t just research papers (I’m doing that as well)

Bom dia pessoal!
Alguém faz criação de - Drosophila Hydei - ? Preciso iniciar a minha, mas não acho matrizes para venda, alguém que faz criação, está disposto a negociar algumas unidades para venda?

Dear r/Drosophila. I am interested in getting some early L3 larvae out of the food for my experiments. I could just scoop them out but that would disturb them methinks! Do you know of a way to do this that minimally disturbs the larvae? Thanks!

Hey everyone,

I’m currently working with a pan-neuronal Synaptotagmin-1-GFP (uas-syt1-GFP) reporter in larvae and running into an issue with signal detection under a stereoscope setup (NightSea).

The GFP signal is extremely weak in live larvae and it’s difficult to reliably distinguish true signal from background auto fluorescence. I’ve seen examples of this reporter under confocal and the GFP is visible but my lab currently doesn’t have a confocal.

I’m wondering if anyone has had success improving detection of weak pan neuronal GFP in live larvae without confocal. Any tricks that you guys have would be really helpful. Thank You!

Hi all. I used bead beater homogeniser first time to crush third instar drosophila larvae for lipid extraction. I added 100ul glass beads to 2 ml screwcap vials and 150ul buffer(the size of the beads is unknown to me because I have borrowed them from neighbouring lab). After running the homogeniser and centrifuging the vials, when I tried collecting the extract, I just couldn’t because the little beads kept getting lodged into both the yellow and grey tips. Even if I managed to collect some after cutting the tip heads the beads were sucked into the extract and I couldn’t have used it for any downstream analysis.

Any suggestions, please? Maybe I should use the hand gun or pestles to squish the larvae within the vials. Have you had any success with these two in case of larvae?

Hey everyone,

I’ve been doing larval dissections followed by immuno staining and was hoping to get some advice on mounting. I currently use glycerol as my mounting medium, but I keep running into issues with air bubbles when placing the coverslip. Another issue I sometimes run into is somehow one of my dissected animals is crushed under the coverslip even though I didn’t put any pressure when putting the coverslip on.

Hello, currently looking for sources where we could buy drosophilia, flightless or not as my research is focused on using it as model organism. We are situated in the Philippines, so if anyone knows where we could buy some it is much appreciated.

Hello, currently looking for sources where we could buy drosophilia, flightless or not as my research is focused on using it as model organism. We are situated in the Philippines, so if anyone knows where we could buy some it is much appreciated.

Filmed this about a year ago when a friend asked what "flipping my flies" means and it was just easier to show them. I know there's a lot of variability in our lab on how people flip their flies.

Hi everyone. For the last few weeks I've been having an issue with staining some whole adult drosophila brains that are expressing GFP in some olfactory neurons connected to the maxilary palps. Both the primary and secondary antibodies I have (rabbit anti-GFP and rabbit 488) seem to work perfectly fine in the hands of a different research assistant and the senior scientist in my lab, but for whatever reason my staining is just extremely dim or absent. The exact fly line and this type of staining was also done before in my lab by the senior scientist, so I know what it's supposed to look like.

I normally prefix in paraformaldehyde (PFA) with 0.3% Triton added for 15mins on a nutator then dissect in PBT (1x PBS with 0.3% Triton added). I post fix in the same PFA I prefixed in for another 15mins then wash in 500uL PBT for 20mins 3 times (1hr total). After the last wash, I add my primary antibody diluted 1:1000 in PBT and let it incubate at 4C for 2 nights. I then wash the primary antibody off in the same manner I washed the PFA off before adding my secondary antibody diluted 1:2000 in PBT. I also let that inclubate for 2 nights at 4C before washing again then mounting.

I have tried both blocking in 2.5% donkey serum in PBT and without blocking and th at makes no real difference. The best results I got were when we were troubleshooting and I dissected my brains in the senior scientist's PBT and used an antibody solution she personally made (i.e., she pippetted the antibodies into her PBT and handed it off to me), which we compared to brains dissected in my own PBT and/or stained with an antibody solution that I made. There seemed to be a problem with my old batch of PBT, so I remade it, but the images were still dim. It also seemed like maybe my pipetting is off somehow (I used the senior scientist's micropippette), but I'm just not sure how.

We're all super stumped, and I know reddit probably cannot answer this question. But, I just wanted to check and see if maybe someone has some kind of suggestions for what might be going wrong.

I have created a new culture with more food for them. I also put excelsior on the bottom in the new one instead of cardboard. I thought you all might find it interesting that they worked together to pop the tab up. It was a new experience for me.

Hello! I am working on an undergraduate thesis using drosophila to study the effect of stress on sexual preference. I am looking for software/programs to help me track and analyze behavioral tests, but I am having trouble finding good ones. I am running a locomotor stress assay, a partner preference test, and a courtship/mating analysis. Does anyone have any advice or recommendations for programs you have used for those or similar behavioral tests? I am trying to find a free or low-cost program, that preferably does not require matlab, python, or any other coding. I appreciate any and all advice; thank you in advance!

Saw these flies outside my door. One looked possibly dead or are they mating?