http://www.scientistsolutions.com/c308-biophysics.html Interdisciplinary field applying techniques from the physical sciences to understanding biological structure and function at the molecular level. Life Science Discussion en-us sci7feed@sci7.nojunkorherepleasespam.com hourly 1 2005-05-05T12:00+00:00 http://www.scientistsolutions.com/t26998-whole+cells+in+patch+clamp.html
We use HEK cells transiently transfected with a K+ channel. It seems that when cells are “driving” a current the seal is always broken before to achieve good a whole-cell. But it works well with HEK cells non-transfected, HEK cells transient...]]> Tue, 28 May 2013 23:48:33 GMT http://www.scientistsolutions.com/t26278-myotonic+dystrophy_+misreg_d+splicing+_amp%3b+gating+of+cav1_1+ca%2b%2b+channel.html ]]> Mon, 18 Mar 2013 09:42:47 GMT http://www.scientistsolutions.com/t26172-unable+to+output+file+from+fluorescence+spectrometer+_td.html I've done a experiment on a fluorescence spectrometer (not the one I usally use) , and I recieved the output in a .td formate. I asked in the lab and a post doc told me the he used spekwin32, to read the data. When I came home I downloaded this program but I'm unable to open the files of interest. Tried every possible way to open them.
I would really love to get down on the data already to night, so if anyone got any alternative programs or any hints on how to use spekwin32...]]> Thu, 07 Mar 2013 09:18:48 GMT http://www.scientistsolutions.com/t25488-cardiomyocyte+patchclamp.html Is anyone here doing patch clamp on cardiomyocytes?
I have a problem isolating viable cells. My cells are too fragmented and I barely get single cells.
I use the enzymes collagenase and Protease and my buffer has MgCl2, KCl, NaCl, HEPES, glucose, CaCl2
Please let me which protocols you use
Thank you]]> Mon, 14 Jan 2013 19:15:55 GMT http://www.scientistsolutions.com/t24565-current%2c+voltage+and+bridge+balance+problems.html So I'm setting up my first rig for intracellular electrophysiology.
I'm using an Axoclamp 2B amplifier and pClamp 9 software, and I have calibrated the software to the Axoclamp. I haven't tried to impale anything yet. This problem starts when I simply place my electrode into the extracellular solution.
My problem is when I inject current down my electrode, the change in voltage occurs in the wrong direction. For example, if I inject a hyperpolarising step to remove series ...]]> Wed, 24 Oct 2012 18:00:49 GMT http://www.scientistsolutions.com/t23734-next+generatiion+science+developed.html FIT or Certainty tool technology developed which is next Generation science.World scientist are welcome to see and test FIT technology.
vk sonakia
Unified Field Scientist
India
vksonakia@yahoo.co.in]]> Sun, 05 Aug 2012 00:26:50 GMT http://www.scientistsolutions.com/t23166-characterization+of+zebrafish+retinal+connexin+hemichannels.html ]]> Wed, 16 May 2012 08:46:29 GMT http://www.scientistsolutions.com/t22981-mycolic+acid.html Sat, 28 Apr 2012 02:42:40 GMT http://www.scientistsolutions.com/t22742-setting+up+an+electrophysiology+rig.html I am in the process of setting up an electrophysiology rig for the first time. Any advice on the best way to go about it. I have all the components, microscope, manipulator airtable etc.
Cheers,
Kwai]]> Mon, 09 Apr 2012 11:11:32 GMT http://www.scientistsolutions.com/t21424-thermal+denaturation+studies.html I was interested in knowing the wide applications of thermal denaturation studies
on proteins. Main application I have read is about stability study of those proteins
used as drugs (vaccine). But other than this if we have thermostable proteins from
thermophilic bacteria, what insight can be obtained from the thermal denaturation
studies. other than comparing with the stability of the mutants. And suppose this protein
is not so relevent ...]]> Thu, 15 Dec 2011 09:57:37 GMT http://www.scientistsolutions.com/t20479-osmometer.html ]]> Sun, 18 Sep 2011 21:55:46 GMT http://www.scientistsolutions.com/t19417-reversal+potential+and+equilibrium+potential.html I've recorded Na+ currents from neurons and I'm trying to fit activation and steady-state deactivation curve.
External sodium conc. was 127mM and internal was 8.6 mM.
ENa is 67.8 mV predicted by the Nernst equation
but actual extrapolated reversal potential was near +42 mV
This deviation possibly arose from contamination outward currents or non-linearity arising form Goldman-type rectification.
Here is my question.
When I fit the activation curve, w...]]> Fri, 01 Jul 2011 11:07:59 GMT http://www.scientistsolutions.com/t18542-reversal+potential+measurements.html I am doing whole-cell voltage clamp in HEK cells and would be very grateful if you share with me your experiences of measuring the reversal potential of an overexpressed channel. How different are the values you obtain between different individual cells that express the same channel?
Theoretically, it should only depend on the ion gradients (set by the bath and pipette solutions) and properties of the channel, and thus should be fairly reproducible from cell to...]]> Mon, 02 May 2011 19:02:27 GMT http://www.scientistsolutions.com/t18340-egta+pka.html Any help with where to find the respective constants?
]]> Thu, 14 Apr 2011 18:17:00 GMT http://www.scientistsolutions.com/t18092-estimating+the+number+of+channels+in+a+patch.html Intuitively I think the number of channels (N) in a patch is:
N=n/p
Being n the average of the maximal levels among trials and p the open probability.
Example 1: I apply a voltage pulse of 500 ms to a patch, many times. I get an average of maximal number levels of 3.32. The open probability is 0.2. Then I got aprox 16 channels in the patch.
Example 2: I apply the same protocol to another patch. I get 0.2 levels. p is 0.1. Then N=2 channels
Correct? Any refe...]]> Tue, 22 Mar 2011 17:04:03 GMT http://www.scientistsolutions.com/t17662-ca+microfluorimetry+in+primary+culture.html I have been using Fura-2 as Ca indicator in islets. Any tip/advise is very welcome...]]> Sun, 13 Feb 2011 17:23:02 GMT http://www.scientistsolutions.com/t17594-fluorsence+problem.html To answer the obvious, I have the wavelength correctly set. The plate is in the ...]]> Tue, 08 Feb 2011 15:31:37 GMT http://www.scientistsolutions.com/t16640-ground+for+noisy+current.html We don't have a real earth ground to get rid of noisy currents. Thus, I used ground of back of amplifier. I connect to copper bend with ground of amplifier then I connect to table, microscopy and digidata. The noisy current is less then 5 pA before I put electrode into the bath solution. However, this noisy current increased up to 20 pA. I try to connect headstage (with wire) to grounded (from amplifier) copper band but it was increased huge n...]]> Mon, 08 Nov 2010 22:59:46 GMT http://www.scientistsolutions.com/t16366-physics.html Sun, 17 Oct 2010 03:25:38 GMT http://www.scientistsolutions.com/t15824-going+whole+cell+problems.html Thu, 02 Sep 2010 12:03:46 GMT