Forster Resonance Energy Transfer, FRET

[kinase]

I'm certain this a long shot but has anybody done FRET before? I'm planning a acceptor photobleach FRET experiment and I'm pretty sure I have everything in working order. I was just wondering if anybody had experience with it and might be able to offer insight on what the best way to do the actual photobleaching is. I'll be using a Nikon A1R+. I'm goign to ask some professors but I assume I just power up the laser and scan the same area a number of times until I observe photobleaching.

I was playing with the scope today. It's a dream come true, as are all the big 4's LSM confocals. I made a great Z stack which is rendered and viewed in 3D of some intestinal lumen. When I do my experiment, I may have a 3D rendering of a nucleus with a few protein interactions. I'll share if I can!

[McConkey]

I have no idea what any of that meant but it sounds awesome! Do you have any images of the equipment you are talking about?
Thanks!

[kinase]

I'm using it as a method to say that two proteins are close enough to potentially be interacting with each other, i.e. within a couple nanometers of each. It's a way to get around the diffraction limit, because I know certain properties of the fluorophores I'm using I can tell if they're close to each other or not.

I wanted to take pictures of the setup but there one someone else there explaining the software to me, I didn't want to look weird . I'll get some next time, when my samples are prepared and I'll probably be the only one in the room. I also recently used a Lecia LMD laser microdissection microscope, I do have photos of that. I'll have to make an equipment porn post one time when I don't have exams looming.