I just started work in a lab where I have to do Gram stains. I'm having trouble with the de-staining step. Everyone has told me it's the most tricky step because it's the only step that isn't timed (the dyes are left on for a specific amount of time, whereas you just have to keep applying the de-stainer until it looks right). Even though everyone acknowledges that it's the hardest part, I'm feeling stupid for struggling so much with it. My boss says I'm de-staining for too long. So I tried some more practice slides where I literally only applied it for a second or two, and STILL, the positive control had parts that were pink. What's more, on a slide where part of positive control was pink, a part of the negative control was purple (i.e., it hadn't been de-stained long ENOUGH). I'm going nuts with this. Any tips on how to de-stain well? Thanks!
I think this video does a good job of explaining the step by step procedures....