From arata from cdb.riken.jp Thu Dec 13 03:11:30 2007 From: arata from cdb.riken.jp (ARATA Yukinobu) Date: Thu Dec 13 13:44:46 2007 Subject: [Celegans] pie-1p::gfp Message-ID: Does anyone have an integrant strain expressing gfp alone driven by the pie-1 promoter? I would like to use for control. Yuki ***************************************************** Yukinobu ARATA, Ph.D Laboratory for Cell Fate Decision RIKEN, Center for Developmental Biology (CDB) 2-2-3 Minatojima-minamimachi Chuo-ku, Kobe 650-0047 Japan Phone +81-78-306-3199 Fax +81-78-306-3200 ***************************************************** From khedges1 from yahoo.com Fri Dec 14 15:52:37 2007 From: khedges1 from yahoo.com (Kathryn Hedges) Date: Fri Dec 14 16:00:20 2007 Subject: [Celegans] Strains with GFP-tagged neurons Message-ID: <994759.6059.qm@web82314.mail.mud.yahoo.com> Speaking of integrated strains with GFP, I'm looking for strains with GFP marking subsets of neurons. I already have ST2 (pan-neuronal), UA57 (DA neurons), and EM800 (YFP/CFP in DA/5HT). I need to check and see if there are plasmids available for RNAi knockdown of DAT-1 or other NT uptake transporters. --Kathryn From khedges1 from yahoo.com Sun Dec 23 09:30:50 2007 From: khedges1 from yahoo.com (Kathryn Hedges) Date: Sun Dec 23 18:20:28 2007 Subject: [Celegans] Fluorescence filter problem Message-ID: <806245.18669.qm@web82301.mail.mud.yahoo.com> This isn't about C. elegans per se, but I got the filters to view CFP/YFP in worms, and I know there are a lot of other folks in the C. elegans community using fluorescence microscopy. I'm probably going to cross-post this to some photography, C. elegans, and microscopy groups because I need help quickly. I bought a set of used dualband fluorescence filters for CFP/YFP (Chroma 51017) on eBay, installed them in a filter cube per the mfr's instructions, and put the cube in the scope. (Zeiss Axioplan 2, the old style with 5 cubes instead of 8.) Instead of seeing a dark field with just the fluorescent cells glowing cyan or yellow, I saw a solid yellow-green field. Same thing if I looked at plain glass slide without anything to autofluoresce, so it isn't just the agarose pad or something. The most likely culprit was the dichroic, so I flipped it over in case I'd installed it wrong. No help. I went through all the combinations of flipping filters and dichroics, exchanging filters, and the only thing that happened is that when I exchanged the excitation and emission filters, the light coming out the lens changed from violet to yellow. (I'm presuming violet is correct because the excitation wavelength needs to be shorter than what the fluorophore emits.) The "outside" face of the exciter looks like someone left a smear when cleaning it, although when I look through it, it looks fine (so the coating is probably OK). Since the set is used, and the seller got it with a used microscope his store purchased from a customer, there's a possibility the dichroic isn't the right one. I hope not... If anyone can help, I'd truly appreciate it. I'm behind schedule for some research I'm trying to present at Model Organisms on Jan. 5-8 (and I have a batch of dual-color worms ready to observe). I forgot to mention this in my other versions of this posting, but Jeff at Chroma has been fantastic. On Thursday I told him about getting this used filter cube from someone else that didn't fit my scope, and he got me the right cube by Saturday--and I'm on the opposite coast from Chroma. --Kathryn