Posted by
Mike Ignatius-2 on
URL: http://confocal-microscopy-list.275.s1.nabble.com/How-to-show-something-inside-the-cell-tp7580451p7580462.html
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Hi Yi,
In a further evolution of the quencher approach consider this paper:
Angew Chem Int Ed Engl. 2013 May 27;52(22):5744-8. doi: 10.1002/anie.201301243. Epub 2013 Apr 19.
A programmable sensor to probe the internalization of proteins and nanoparticles in live cells.
Liu H, Johnston AP.
Instead of bathing the whole prep in quencher, with its associated off target concerns, a complimentary nucleic acid probe is made with a quencher. Much more specific and pretty easy to make.
Mike Ignatius
Marker Gene Tech
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Hi Yi - I think your solution is good. Fluorophores on the surface can also be selectively quenched, depending on the fluorophore - FITC by lowering pH, red fluorophores or those emitting in the 600s by adding several mg/ml of Acid Blue 9.
Mike Model
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Dear listers,
One of our user is investigating the transportation of fluorescence labeled DNA oligo in cells. He would like to prove the DNA oligo has been transported inside the cells but not on the surface of cells by using the light microscopy. We suggested him to clearly label the membrane of cells and use the Z-stack images of confocal to look through the cells.
I am wondering if there is any other better solution?
Any suggestion on good membrane dye is appreciated as well!
Thank you,
Yi