last three days

the last three days I have still been working on dissecting ovaries from drosophila flies and them preparing the slides to visualize them but now I understand a bit more the reason why we are doing this, I say we cause we are in fact about 6 people all doing the same. We set up crosses of flies in order to obtain progeny with all the characteristics that we need. one of the flies has a slbo that as i mentioned before makes it border cell specific and this is linked to gal 4 gal 4 in turn is used to switch on the expression of gfp by UAS and also works to swicch on the dsred that is a red dye with a nuclear localization signal overall the genes in that chromosome help us identify and visualize the cells we are interested in but also we need to make sure they keep these genes so recombination is avoided by having a balacer chromosome. which is a chromosome that has a lot of inversions and avoids recombinations, I have been informed that these fly lines with balancer chromosomes were created by radiation. The balancer chromosome also has another gene that helps us identify what we are looking for and that is it has the cyo gene that makes the wings of a fly curly ( I'll come back to that now). Now the cross also has two chromosomes in one it has the UAS transgene and in the other the cyo. The UAS transgene is what we are interested in because it is the one that we have manipulated to either overexpress or inhibit a kinase or a phosphatase enzyme, the other is used to screen the flies, that is if the flies have curly wings they have inherited the chromosome that we are not interested in whereas if they have straight wings they have the UAS transgene.
What these means is that we have to screen these flies and there are so many of them, they are all labeled with the different kinase or phosphatase that has been altered and when we dissect the ovaries and fix them into a slide it is then possible to screen the slides looking for alterations in migration and movement. The past three days I have been improving my dissecting technique as well as my fixing technique. I really struggled getting the slides to a good standard with a little background that allows the visualization of the slide but I have definitely improved specially today that I manage to produce really good and clear slides. I suppose the key was to allow the PBS to rinse for a longer time !!!!! I am getting there and I am getting better at it !!!!