I am trying to determine the best possible method to measure the impedance on my monolayer of MDCK (Madin Darby Canine Kidney) epithelial cells.
Does this require a specific metallic coating (via sputter coater or another method) on/ under (as a membrane the cells would grow on) my monolayer or would the liquid media itself simple provide the conductive element required?
Hi Divya,
can you give more details to your system, please? What kind of liquid media do you mean? Do you have SAMs and cells both?
Thank you,
Kirill
Water will do the work. The Dielectric constant is about 85.5
I think the best method would be to grow your cells on a permeable support and meassure impedance in a ussing chamber. Hope this can be of any help! Cheers!.
Can you give us some more details about what you want to determine? Which is the reason for you need the impedance of a cell monolayer? What parameter do you intend to evaluate and for which specific use?
Contact Dr. Jeffrey Fredberg, Professor of Bioengineering and Physiology [email protected] and/or Dr. Enhua Zhou, [email protected] and/or Dr. James Butler, Senior Lecturer, all at the Dept. of Environ. Health, Harvard School of Public Health, Boston; they are real experts in this field of work
thanks guys :)
@Kyrylo: I'm growing my cells in DMEM (Dulbecco's Modification of Eagle's Medium) with a high glucose concentration. The cells form a monolayer that adheres to the bottom of the dish
@Daniel: I'm afraid of electrocuting the cells. This may seem childish, but it seems like a legitimate concern at this stage
@Edith and Stephania: thank you. I'll look into this technique
@Javier: yes, I do believe you are right. I am considering building a chamber with the permeable support being PDMS (the simplest thing I can think of). I am trying to figure out the best way to perforate the PDMS to create holes as small as possible. I want to target 10 microns. Not sure whether to use lithography or just punch it. Do you have any suggestions?
@Lucia: the experiment is titled "trafficking of nanoparticles through an epithelial membrane". I want to visualize, using confocal microscopy, the passage of nanoparticles through (or whether it merely activates a process) the epithelial membrane. While I have not yet gone as far as determining which NPs to use, I am trying to create a monolayer to ensure that the NPs will not fall through any holes that may be created. The impedance measurements should provide some data as to the integrity of this monolayer. I can't seem to think of another way and welcome alternate suggestions to impedance measurements.
@Peter: thank you sir. I will look into this suggestion
When I measured the impedance of epithelia (I think it was South African frog - the tissue lives long) with my students, we did not attach any electrodes, since as Danielo Olapado mentions, the conducting (sea) water is a "good enough" electrode. The epithelia has been stretched in the middle of a sample cell with two Pt electrodes on each side of the stretched epithelia. Proper and calibrated Impedance measurements have enabled us to identify various conducting channels through the epithelia, inclusive a quantitative analysis of Calcium passive channel. The number of the elemental Ca channels did agree with electron microscope studies of the same. Yes, the impedance measurements are a good qualitative and quantitative characterisation tool. With best regards
Petr
Our company develops 3D cell barrier culture systems for in vitro study of virtually any cell barrier, and the system comes equipped with both a Fluid Perfusion Unit and a Trans-Endothelial/Epithelial Electrical Resistance (TEER) Measurement Unit! The use of a vessel-shaped 3D environment has been proven more effective than 2D systems like Transwell (
Article Santaguida S, Janigro D, Hossain M, Oby E, Rapp E, Cucullo L...
), and our advanced TEER Measurement Unit allows for frequency sweeps between 0.1-1,000 Hz, automated time point sampling, logging of data to Excel, and additional measurement of phase angle for cell capacitance calculations! For those that are committed to Transwell use, we also have a TEER Measurement Unit that is compatible with nearly all Transwell products (Endohm cell cup chamber, STX2 "chopstick" electrodes, etc) and allows the user to greatly expand their testing capabilities with Transwell equipment. Additionally, we have smaller modular systems that connect to microscope-friendly cell culture modules: i.e., a miniaturized 3D cell culture system that you can use right under your microscope! If you or anybody on this thread is interested in learning more, I encourage you to visit www.flocel.com or email me at [email protected], thank you!
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