I don't know the precise answer off-hand, but I do recall that the duration depends on the size of your pipette tip, so roughly depends on pipette resistance. Below I include several references where you should find a useful approach to calculating the time constant for your conditions.

Oliva, C., Cohen, I. S., and Mathias, R. T., 1988, Calculation of time constants for intracellular diffusion in whole cell patch clamp configuration, Biophys. J. 54: 791–799.

Pusch, M., and Neher, E., 1988, Rates of diffusional exchange between small cells and a measuring patch pipette, Pflügers Arch. 411: 204–211.

If you can find a copy of the book "Single-Channel Recording", then you should find the answer in this chapter: "Tight-Seal Whole-Cell Recording" by Marty and Neher. Chapter Tight-Seal Whole-Cell Recording

Hope this helps - let us know what you calculate!

Hi Is good question, but in depende of the size of the cells or the capacitance of your model. In personal work we waiting a hours for have a change in the intracellular composition.

But, remember in whole cells configuration you only patch a part of the cells, not complete, and Cs solution not controler all the channel inside. Maybe you need considerate a external solution

Depends on the size of your pipette tip (i.e. electrode resistance) and the size/shape of the cell. I would say anywhere between 5 min (for the cell body) and ?? min for the distal dendrites. If you need very precise control, you'd have to check experimentally as Timal says.

Early 80s Alain Marty and Berth Sackmann measured how long it takes for the new Cl equilibrium to take place in the cells they changed. I rememberer it was surprisingly quick, in the order of the minute.

Hi, I usually start my measurements in HEK cells after waiting for about a minute. That should be enough time for the internal solution to equilibriate within the cell. My pipette resistance is usually around 1.5 to 3 MOhm.

Please see the followiing papers:

https://www.ncbi.nlm.nih.gov/pubmed/?term=pusch+Neher

There is an instrument available for whole cell perfusion

http://www.npielectronic.de/products/perfusion-systems/ala-2pk-whole-cell-pipette-perfusion.html

This is an improved version of the model described in this paper

Article Perfusing patch pipettes

Exchange times that have been opttained are in the range of about one minute!

This is in good agreement with the paper Mathias et al Biophys J 1990 (theory)

https://www.cell.com/biophysj/pdf/S0006-3495(90)82418-8.pdf

and the paper by Push and Neher Pflüg. Arch 1988 (experimental)

Article Rates of diffusional exchange between small cells and a meas...

Hope this helps!

Hans Reiner Polder

Hello, I agree with Johanes and Felix. In my experience with pipettes of arround 2-3 MOhm an inhibitor or blocker can take 2-5 minutes to reach the maximum effect, and an ion that is very much smaller takes less than a minute.

It's like Timal wrote, measure the speed of GABA reversal potential stabilization. Alternatively, with another chloride channel that is expressed in your cell type.

My experience is as the following:

1) For small cells, like HEK cells, most of intracellular solution replacement by the pipette solution occurs within 2 to 3 min;

2) For larger cells, like adult rabbit ventricular cardiomyocytes - for about 5 min;

3) For remote neuronal processes, like dendritic spines of CA3 rat hippocampal neurons - from 15 to 30 min.

This is valid only for membrane impermeable molecules and ions, while permeable ones reach intermediate submebrane steady-state concentrations.

Wait 3 minutes minimum before starting your recording

Depends on pipette resistance, cell size and diffusion koefficient for your molecule. For fluorecent Fura its less than 10 min to get to spines.

I think for ions it's faster than for for the dye.

I did cell filling for post analysis and most retinal ganglions would be filled in 10 min