Is it necessary to have a special microscope filter to visualize transient transferred mKeima cells? The microscope I have been using has the typical GFP and mCherry filters, but not a special filter to see mKeima fluorescence.
Dear Emily Fr ,
You will know, that mKeima is a fluorescent Protein to measure the intracellular pH. As you can find here:
https://www.fpbase.org/protein/mkeima/
Excitation should be around 440 nm and you will have to measure two emission wavelengths i.e. 650 nm and 600 nm, but I have never used mKeima. Standard GFP and mCherry filters will not work, since a GFP filter might be able to excite your protein the band pass filter for the emission will not match the emission of the mKeima. And mCherry Filters will not excite your protein although they should have the right emission filter. If you have the possibility to use a long pass filter for the emission you might be able to check the transmission. Or you can try to use the emission filter of the mCherry Filter set in the GFP Filter set instead of the existing GFP Emission Filter, but that might depend on the dichroic filter.
Best wishes
Soenke
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