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
> ================================================
>
>
|
