From anjum_zfr from yahoo.com Sat May 10 09:23:31 2008 From: anjum_zfr from yahoo.com (anjum_zfr@yahoo.com) Date: Sat May 10 14:03:16 2008 Subject: [Neuroscience] Re: Technical question: dissociated cells References: <07060602-51f4-4be0-a683-ec8ce45cae11@b5g2000pri.googlegroups.com> Message-ID: <643383b1-4827-4b56-a7f3-06a37407a924@q27g2000prf.googlegroups.com> Hi , we are using newborn hipppocampal neurons for patching at JCSMR, ANU, Canberra. they grow well and stick to glass coverslipes as well. Though I am new student here and learning patching here. I don't have the references with me rightnow but i can send you them if you wish. Cheers Anjum J.A.Legris wrote: > On Apr 24, 2:15?am, christinela...@hotmail.com wrote: > > Hi, I'm wondering if anyone here has done electrophysiology on acutely > > dissociated neurons? I've found several methods in the literature and > > have had a couple of attempts with adult rat brain. The cells come out > > looking bright and round and alive, but unfortunately I can't make > > them stick to glass chips, with or without poly-D-lysine coating. I > > need them to stick to something in order to patch them. > > > > Any suggestions? > > How about a micropipette under low negative pressure. > > -- > Joe From edzska from gmail.com Sun May 11 21:17:03 2008 From: edzska from gmail.com (Ewa Z-ska) Date: Mon May 12 12:00:18 2008 Subject: [Neuroscience] Re: Technical question: dissociated cells In-Reply-To: <643383b1-4827-4b56-a7f3-06a37407a924@q27g2000prf.googlegroups.com> References: <07060602-51f4-4be0-a683-ec8ce45cae11@b5g2000pri.googlegroups.com> <643383b1-4827-4b56-a7f3-06a37407a924@q27g2000prf.googlegroups.com> Message-ID: <3bee93df0805111917u70cecbeej831bba379f3aac8a@mail.gmail.com> Hi, You may want to try approaching your cells with low positive pressure. Test if they are sensitive to the pressure in your recording pipette. Cells should stick to the poly-lys coverslips. Give them a few hours to do it so. best, eva On Sat, May 10, 2008 at 9:23 AM, wrote: > Hi , > we are using newborn hipppocampal neurons for patching at JCSMR, ANU, > Canberra. they grow well and stick to glass coverslipes as well. > Though I am new student here and learning patching here. I don't have > the references with me rightnow but i can send you them if you wish. > > Cheers > Anjum > > J.A.Legris wrote: > > > On Apr 24, 2:15?am, christinela...@hotmail.com wrote: > > > Hi, I'm wondering if anyone here has done electrophysiology on acutely > > > dissociated neurons? I've found several methods in the literature and > > > have had a couple of attempts with adult rat brain. The cells come out > > > looking bright and round and alive, but unfortunately I can't make > > > them stick to glass chips, with or without poly-D-lysine coating. I > > > need them to stick to something in order to patch them. > > > > > > Any suggestions? > > > > How about a micropipette under low negative pressure. > > > > -- > > Joe > _______________________________________________ > Neur-sci mailing list > Neur-sci@net.bio.net > http://www.bio.net/biomail/listinfo/neur-sci > -- Ewa From ytf0707 from gmail.com Mon May 12 00:46:59 2008 From: ytf0707 from gmail.com (ytf0707@gmail.com) Date: Mon May 12 12:00:23 2008 Subject: [Neuroscience] Silver staining......quick protocol!!! rather Bielschowsky's method Message-ID: How about an immerse in Silver nitrate solution (10%) for 15 min take the slides out and wash with distilled water and then add Ammonium Hydroxide solution (1%) to be just clear put the slides back, stain for 30 min then JUST wash with distilled water? Do you think this works? Anyone has an idea on the neurochemical bases for this reaction/ staining? Or anyone can introduce me some quick protocol for silver staining? From tehgabriel from web.de Fri May 16 03:44:06 2008 From: tehgabriel from web.de (Tom) Date: Fri May 16 11:13:40 2008 Subject: [Neuroscience] Re: Technical question: dissociated cells References: <07060602-51f4-4be0-a683-ec8ce45cae11@b5g2000pri.googlegroups.com> Message-ID: <04179506-03a7-4a16-9915-28b84286dfcb@b64g2000hsa.googlegroups.com> On Apr 24, 8:15 am, christinela...@hotmail.com wrote: > Hi, I'm wondering if anyone here has done electrophysiology on acutely > dissociated neurons? I've found several methods in the literature and > have had a couple of attempts with adult rat brain. The cells come out > looking bright and round and alive, but unfortunately I can't make > them stick to glass chips, with or without poly-D-lysine coating. I > need them to stick to something in order to patch them. > > Any suggestions? - Make sure your glas cover slips are really clean (standard protocols are available in the internet. briefly: sonicate coverslips in a glas beaker with nitric acid for several hours, rinse with milliq water to remove the acid, rinse with ethanol, sonicate with ethanol for another several hours, rinse with fresh ethanol and finally store them in ethanol. protect cover slips from getting dry at any time point!) - Try different coating substrates. e.g. d- or l- lysine, or a mix of collagen and lysine (we use something like 1ml of milliq water with 100 ul rat tail collagen plus 20 ul PDL). in my experience, postnatal neurons (mouse) don't grow so well on pure PDL coating. - Check your medium! there are plenty of different receipes how to prepare culturing medium. In general they should contain some kind of supplement (N2 or B27) and some serum (horse serum or fetal bovine serum), diluted in the medium (MEM, neurobasal, etc.) we use (for 100 ml final volume): 87 ml neurobasal, 10 ml horse serum, 2 ml B27 supplement, and 1 ml glutamax. If you want/need you can add antibiotics. This works fine for our postnatal mice hippocampal cultures. Regards, Thomas From CFenster from wooster.edu Sat May 17 14:28:48 2008 From: CFenster from wooster.edu (Catherine Fenster) Date: Sun May 18 12:30:44 2008 Subject: [Neuroscience] Analog-to-Digital Conversion For Sale&In-Reply-To= Message-ID: <482EF9AF.5DD1.00F9.0@wooster.edu> Hi Ronald Chase, I am interested in your AD board. Here is the catch...I need it for a research sabbatical (for only 6 months). This means that I'd either like to borrow/rent it or purchase it with a discount. How much were your planning to ask for it? Any chance that I could convince you to lease it??? Thanks for your help. -Cate Fenster *************************************** Catherine (Cate) Pichon Fenster, Ph.D. Assistant Professor of Biology Department of Biology (Mateer 309) The College of Wooster 931 College St. Wooster, OH 44691 office: (330) 263-2436 fax: (330)-263-2378 email: cfenster@wooster.edu *************************************** -------------- next part -------------- BEGIN:VCARD VERSION:2.1 X-GWTYPE:USER FN:Catherine Fenster ORG:;Biology EMAIL;WORK;PREF;NGW:CFenster@wooster.edu N:Fenster;Catherine END:VCARD From connelly.bill from gmail.com Sun May 25 17:51:51 2008 From: connelly.bill from gmail.com (Bill) Date: Sun May 25 20:03:26 2008 Subject: [Neuroscience] What camera do you use of Fluorescent Imaging? Message-ID: Hi, I need a new digital camera suitable for a variety of tasks in epifluorescent mode (Plain luminescence calcium imaging, ratiometric calcium imaging as well as plain old 'don't glowing cells look pretty' mode), so I'd need to be able to change gain and exposure time at a click of a button. On top of that, in order to dovetail to electrophysiological recording, I'd need it to be easily triggered into action by an ouput from the A/D converter. I've played around with the Diagnostic Instruments Spot Explorer line of cameras, and while they have a good set of functions, some bright spark had mounted a fan in the camera and hence it generated lovely vibrations that constantly shook the rig. I was just wondering what camera/acquisition software do you guys use? Thanks Bill Connelly Department of Pharmacology University of Otago New Zealand From christinelaura from hotmail.com Sun May 25 18:22:20 2008 From: christinelaura from hotmail.com (christinelaura@hotmail.com) Date: Sun May 25 20:03:41 2008 Subject: [Neuroscience] Re: Technical question: dissociated cells References: <07060602-51f4-4be0-a683-ec8ce45cae11@b5g2000pri.googlegroups.com> <04179506-03a7-4a16-9915-28b84286dfcb@b64g2000hsa.googlegroups.com> Message-ID: <249792cf-9902-40f2-a444-fd3d5d9db3fe@x19g2000prg.googlegroups.com> Hey thanks for all the suggestions! I've tried lower/no positive pressure and still ended up chasing cells around the dish. I shall try leaving them on the chips for longer and maybe different chip coatings next. Thanks again -C