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What is STEP and What is RAMP in electrohysiology protocol [View Printable]
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neurons
Group: Member Posts: 3 Joined: Sep 17, 2008
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I am doing my Ph.D in Neuroscience(Electrophysiology), but my background is not in this area , I am using AXON instrument so can anybody tell me in that when u design a protocol in the wave form option it says STEP or RAMP what does it mean what exactly happens when u put step and when u put RAMP
how can one know that this is a suitable protocol for ur type of cells or ur specific conditions
If there is any very basic book which can clear my doubts then please let me know, like when one patch embryonic neurons and when u don't put the EGTA, or sodium what happens like i want to know certain basic requirement as one has to add K+ to depolarize the membrane so how do u know what is function of all the component of ur extra and intra and why it is being used at particular concentration only
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Posted Sep 17, 2008, 11:37 AM |
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frasermoss
Group: Admin Posts: 724 Joined: Feb 22, 2005
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Firstly some essential reading
The axon guide can be downloaded here and has plenty of useful pointer for writing protocols
Also I frequently recommend this book as a plainly written guide for beginners
Patch Clamping: An Introductory Guide to Patch Clamp Electrophysiology by Areles Molleman # ISBN-10: 047148685X # ISBN-13: 978-0471486855 here is the link to the book on Amazon.com
RE the step vs ramp protocol question
Ramps have the advantage of generating current-voltage relations directly and very rapidly and might be considered suitable to study for a study of rapidly activating or time independent currents.
Step protocols measure the steady state current at a given voltage and depending on the duration of the step allow the investigator to also analyze the kinetics of the current at each voltage and view phenomena including inactivation, desensitization at a particular voltage
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......................... "Opportunity is missed by most people because it is dressed in overalls and looks like work". Edison
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| Posted Sep 17, 2008, 13:02 PM |
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neurons
Group: Member Posts: 3 Joined: Sep 17, 2008
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thanks a lot it really cleared my basic doubts
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| Posted Sep 18, 2008, 3:28 AM |
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neurons
Group: Member Posts: 3 Joined: Sep 17, 2008
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FRASER MOSS Sir I have one more request if you can help me in your published paper Proc. Natl. Acad. Sci. USA Vol. 95, pp. 8328–8333, July 1998 in MATERIALS AND METHODS It is mention as The pharmacological sensitivities of the 10–13 (a1A-2, a2bd, b4a), G1A1 (a1B-1, a2bd, b1b), C11D8(a1C-1, a2bd, b2e), 5D12–20 (a1D, a2bd, b3a), E52–3 (a1E-3, a2d,b1b), and E58–19 (a1E-3, a2d, b4a) cell lines, encoding P-Q, N,L, and R-type channels, respectively, have been reported (17).
what are this cell lines and sorry but the reference I am not able to get Boot, J. R., O’Brien, A. & Tran, S. (1997) Brit. J. Pharmacol. 122, Suppl. 1, 89P (abstr.).
If you still have soft copy of this refernce then please send it to me
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| Posted Sep 18, 2008, 7:27 AM |
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