Wouldn't it be funny if you had to reengineer the patch clamp apparatus to provide positive pressure to keep the cell out of the micropipette, while simultaneously providing a leak of chemoattractant so that the cell's own pressure maintains the seal. 

But that's speculation. 


On Jul 25, 2012, at 1:30 PM, Thierry Soldati wrote:

> … and because I love to agree with Rob, I would add that if you want to measure plasma membrane channels, his suggestions are cool.
> I was more thinking in terms of capacitance measurements to monitor membrane traffic !!
> Cheerio,
> 
> Thierry
> 
> 
> On 25 Jul 2012, at 19:44, Rob Kay wrote:
> 
>> Dear Wanessa,
>> 
>> I love to disagree with Thierry, and so I would give latrunculin a go (at
>> least for one Friday afternoon)!
>> 
>> As a non-druggy alternative, you could also try either of our
>> temperature-sensitive membrane trafficing mutants: nsfA or secA (see
>> papers in J Cell Sci by Traynor et al and Zanchi et al).  At the
>> restrictive temperature of 27oC these mutants round up and stop moving
>> within a few minutes, and I think they would then be easy to patch clamp.
>> 
>> If any of these tricks worked you would be doing stuff that almost nobody
>> has been able to do before......
>> 
>> Best wishes,
>> Rob
>> 
>> 
>>> Hi Wanessa,
>>> 
>>> In one word … don't try !
>>> When I came to Heidelberg, I was hired by Wolfhard Almers, one of the very
>>> top electrophysiologist, because he wanted to use Dicty to measure the CV
>>> discharge … they tried for 2 years !! Never worked. Dicty either crawls
>>> around the pipette, or inside, but there is no way to calm it down. I
>>> would not try pharmacology to "tranquilise it".
>>> Cheers,
>>> 
>>> Thierry
>>> 
>>> On 25 Jul 2012, at 16:40, Wanessa Cristina DE LIMA wrote:
>>> 
>>>> Dear all,
>>>> 
>>>> 
>>>> A professor in my department, specialized in electrophysiology and
>>>> patch-clamping, agreed to give it a try and patch-clamp Dictyostelium.
>>>> Although I’ve never seen this described in papers, we decided to try
>>>> anyway. She thought the cells would wrap themselves around the
>>>> electrode, or just run away. However, to her astonishment, Dicty cells
>>>> were able to ENTER on the 1-micron-pipettes used to patch clamp.
>>>> 
>>>> 
>>>> So, my question is: is there any trick around to stop cells and still
>>>> have them in a “physiological” condition? I was thinking latrunculin,
>>>> but not sure if it’s the best answer….
>>>> 
>>>> 
>>>> Cheers, Wanessa
>>>> 
>>>> 
>>>> Lab Pierre Cosson, Geneva
>>>> [log in to unmask]
>>> 
>>> --
>>> ================================================
>>> Prof. Thierry Soldati
>>> Department of Biochemistry
>>> University of Geneva
>>> 30 quai Ernest Ansermet, Sciences II
>>> CH-1211-Genève-4, Switzerland
>>> 
>>> Tel: +41-22-379-6496
>>> Fax: +41-22-379-3499
>>> email: [log in to unmask]
>>> http://cms.unige.ch/sciences/biochimie/-Thierry-Soldati-.html
>>> ================================================
>>> 
>>> 
> 
> -- 
> ================================================
> Prof. Thierry Soldati
> Department of Biochemistry
> University of Geneva
> 30 quai Ernest Ansermet, Sciences II
> CH-1211-Genève-4, Switzerland
> 
> Tel: +41-22-379-6496
> Fax: +41-22-379-3499
> email: [log in to unmask]
> http://cms.unige.ch/sciences/biochimie/-Thierry-Soldati-.html
> ================================================
>