I found a dye on the Internet, but I do not know how this is work: CellMask Actin Tracking Stains. Somebody used that dye before? Is it good for tracking the actin filaments or which dye could be the best for this experiment?
Thank you for your help and answears.
I dont know much about Actin dyes for staining live cells, but have you considered expressing GFP/RFP tagged actin ? Most stains are toxic to live cells and may interfere with your observation. If you can transiently transfect a flourescent labelled actin construct before your experiment, you can make your observation with little or no damage to the cell.
Hi there,
I haven't used this particular dye, but the webpage of the company reads:
"CellMask actin tracking stains detect F-actin with a targeting structure that closely resembles jasplakinolide [...]"
So, you can assume that the mechanism of action is going to be similar to that of jasplakinolide... meaning that probably interferes with actin dynamics (by promoting and stabilizing actin polymerization).
There is an interesting document that compares several method to visualize actin in live cells and compares their advantages and limitations, find it here:
https://jcs.biologists.org/content/joces/130/3/525.full.pdf
The idea suggested above about GFP-tagged actin could be suitable for your experiments, although it has been also described to have an effect over actin dynamics. There is another alternative, which is LifeAct-TagGFP2, this is not actin, but a recombinant Actin Binding Protein fused to GFP, although there are also some papers suggesting a certain effect over actin dynamics.
See the links below for a couple of references:
https://en.wikipedia.org/wiki/LifeAct_Dye (Just to find the link to the company that sells it)
Article Differential effects of LifeAct-GFP and actin-GFP on cell me...
https://pubmed.ncbi.nlm.nih.gov/31239446/
Cheers,
J
Thank you for your answer. Yes, I considered that option too, but I would like to use a dye which can be used in live cells and I read about this CellMask Actin Tracking Stains and it is very promosing, but I could not find any personal opinion about this dye. I will keep in mind this tagging option.
J. Ramirez-Franco Hi!
Thank you for your answer. I agree with you and Praveesh Valissery , maybe the tagging is the best option. I am going to read the documents what you send.
Have a nice day,
Loretta
As Praveesh stated, if you can transfect cells, that will likely give you great staining of the F-actin. You could even consider using LifeAct in place of FP-actin so you are not over-expressing actin itself (either way, it provides very high quality F-actin labeling live [especially so if cells are fixed]). My concern with this dye you mention is that it may not look anywhere close to as good as transfections /phalloidins and as such may end up just being terrible when viewing under the microscope. I have never tried these actin-dyes for use in live cells before so maybe someone who has can comment on their quality of staining actin versus transfection/phalloidin.
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