Patch clamp technique in India ?

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Sathya Subramani

We run a patch-clamp lab at CMC, Vellore.

Ajay Kumar

Thanks dear Sathya,

But the problem for there is that it's a 37 hr run from chd to vellore and I need to maintain the potential in live cells. So maintaining the viability during such long run is a big issue..

As you have an experience in the same field, any more valuable suggestions are highly welcomed..

De-Shou Cao

Calcium fluorescent imaging is the easiest way to measure the function of the cells. The calcium indicator, flu-4, can be used. A microscope attached with a CCD camera can monitor the fluorescent intensity change using micromanager and Image J (free software).

You could do your cultures here and then patch the cells.

@ De-Shou Thanks dear.

Can you tell the principle and how applicable is this dye in measuring voltage potential and how reliable these results can be as compared to Patch clamp measurements?

Thanks !

But coming there, doing cultures and then inducing the differentiation will take another one and a half month and then we will be able to measure potential. I am in the final stage of sending the article for publication . Second I have to see potential in 4 different type of stem cells and BMSCs will not survive the travelling time and cryopreservation conditions. Is there any other alternative you can suggest ?

Thanks in advance..

Norbert Weiss

An alternative is to use voltage-sensitive dye to measure cell electrical signal. It will not be easier than using patch-clamp but if you already have imaging technology on-site it might be something to consider depending of what you want to measure / characterize exactly. Please have a look at this to have an idea http://eknygos.lsmuni.lt/springer/374/Part%202/3/5%20Item.pdf

Thanks dear Norbert,

We have an access to very advance imaging techniques including confocal and EM..so Imaging technology will not be a hurdle..

Jitendra Narain Singh

we also involve in patch clamp study. We are basixcaaly involve in voltage and current clamp recording

Thanks dear Jitendra,

May I have your contact details please because NIPER is quiet close to PGI and may be I can visit sometime?

Cheers!

[email protected]

Ekta Mayur Dembla

Hi Ajay,

You can measure the calcium increase in the cell through ratiometric dyes like Fura-2, Fura-2F, Fura-4F, Furaptra (for quantitative measurements by which you can meaure the calcium concentration increase you can use the combination of above mentioned dyes according to the range of calcium concentration that these dyes cover). If you need only qualitative measurement just to see whether calcium is increasing or not you can also use Fluo-4.

Regards,

Ekta

Thanks Ekta....Can you Please send some suitable link for the same describing protocol so that I can go into the detail of the process and see wheather it solves my purpose or not..I need a relative quantification method as I want to compare the efficacy of neural differentiation between different stem cell types.

I am sending you one article about intracellular calcium measurements. You can go through it and then let me know if you need more details.

Thanks dear Ekta...

Ramachandra Nagaraju

Dear All,

I have a yeast S.cer clone expressing recombinant ion channel protein localized to the plasma membrane.

Unfortunately, we do not have a facility for Patch clamp assay to check the function of an ion channel protein.

I request your kind suggestion and help to perform the whole cell patch clamp assay for yeast system. If not patch clamp, could please suggest me any other methods to check the function of an ion channel expressed in a yeast host.

Your help and suggestion will be greatly acknowledged.

Ramachandra