Anyone has experience with a truly permeable DNA stain that will not be adsorbed in plant cell walls? We work on Arabidopsis roots and we tested dyes that are membrane permeable in animal cells but we had no luck with our material. In protoplasts there is no problem, therefore it seems that the obstacle is the cell wall charge (or something related)
Dear George, for the observation of the living nucleus you can try H2BGFP lines.
Please, consider that you stain majority of DNA staining is intercallation dye, so, they may have strong effect on gene expression. Please, also consider that majority of the dye cant penetrate membranes of the living cells (DAPI, Hoechst etc). So, may be histone-GFP labeling will be the best option.
If you're not afraid with green stuff try SYBR-Green I and II or Syto 9
If you need to see GFP or other green things you should try Syto Orange 83. It stains really well nuclei of Medicago roots at 5µM and it does not impair root growth when added in the culture medium (tested for 2 weeks).
There are also Syto Red(s) but I haven't tested them.
Hi George,
you can also look at blue stain:
Hoechst 33342
https://www.thermofisher.com/order/catalog/product/62249
http://www.bio-rad.com/webroot/web/pdf/lsr/literature/10043283.pdf
not sure how it works in plants,
best,
Jozef
Hi George,
I stained leaf cuttings and acridine orange worked well. If you want to try I can offer you some.
Saša
Hi George,
for my experience in plant cells the DAPI stain DNA in live cells very well, however the some detergent ( I use 0,1% Triton 100) must be included in the protocol. Thus, if the detergent does not interefere in your experiment try to use them as I have recommended. Good luck!
Thanks a lot all of you for your answers.
Olivier: I should definitely try this thanks for input. I will
Jozef: Yes this should be cell permeable so it is worth a shot. I will put it in order
Alexandra: This is nice alternative. I think we have some in the lab. If not I will contact you
Vladimir: We also tried TX-100 but our samples die. For us it is essential to maintain nuclear staining over a period of hours and keep our cells alive
Dear George, for the observation of the living nucleus you can try H2BGFP lines.
Please, consider that you stain majority of DNA staining is intercallation dye, so, they may have strong effect on gene expression. Please, also consider that majority of the dye cant penetrate membranes of the living cells (DAPI, Hoechst etc). So, may be histone-GFP labeling will be the best option.
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