Hello,

I'm a Msc student who is working on some Drosophila Melanogaster Bicoid Homedomain mutant lines, I did molecular staining on embryos which is tagged for Bicoid and obtained Confocal images of several different mutant lines. Each mutant line has been bred in two different temperatures which are 22C and 25C. I want to know how Bicoid concentration gradient has shifted for each temperature. I'am using Fiji (ImageJ) Plot feature and obtaining the data for each image, but I just couldnt figure out how to effectively compare two different temperature to get a meaningful result. I've researched and find some articles about this and tried to calculate the point in Embryo Length where the Bicoid concentration dropped and reached to a certain intensity level which is called lambda (1/e), but it gave me inconsistent results, I just need some advice on how to compare the differences caused by the temperature in Bcd gradient, would love to hear from someone who's worked on calculating the Bcd gradient in differently conditioned embryos. Thank you so much!