Hey I am about to start drug survival test in drosophila does anyone has tips of how to select drug concentrations, fly density.
And also do you put either paper filter or other kind of filters in your adult cultures as a way to increase the surface for the flies? If so do you change the paper if so how often?
Thank you!
Drosophila survival assays
For intestinal infection, survival was assayed as previously described (Blow et al., 2005). Briefly, 30 male flies were divided evenly between three vials either containing a cellulose acetate plug saturated with 2 ml of LB broth supplemented with streptomycin alone or containing a 1:10 dilution of an overnight culture of V. cholerae that had reached early stationary phase. Viable flies were counted at least once in 24 h. For septic injury, 20 flies were pricked in the dorsal thorax with a 29-gauge needle dipped in a solution containing early stationary-phase bacteria diluted to an OD600 of 0.05 in fresh LB media. Flies were maintained in vials containing a cellulose acetate plug saturated with 5% sucrose in phosphate-buffered saline (PBS). Flies were inoculated in the evening, and the numbers of viable flies were recorded hourly during the following day. Survival curves were constructed for both intestinal and systemic infections, and log-rank tests were used to determine statistical significance. Reproducibility of all survival data was confirmed in at least three independent experiments.
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2666625/
You can use filter paper soaked in 5% sucrose solution(or depending on what recipe you use, appropriate sugar content) and the drug of your choice. This is typically used in paraquat stress tolerance assays. The filter paper can be cut to specifications using scissors, inserted whole, or cut in 1/4 quadrant-circles and inserted into the vials. The required amount of liquid then can be added.
My personal preference is using 1/4 of a filter paper and soaking it ~300ul of the solution. That provides enough moisture that the paper sticks, the liquid doesn't flow and flat enough that flies can survive on it.
Ideal fly density is same as that standard fly rearing conditions i.e.
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