In this context, our group want to deliver a chemical compound inside dendritic cells. However, to confirm successful delivery needs to marker this compound with a fluorescent marker visualizing the delivery and release of this compound at the target place, as well as facilitating the tracking of these cells. Thank you
The easiest way, if you are not going to do the synthesis from scratch, is to attach the fluorescent dye at an amino group, a thiol group or a carboxylic acid group using commercially available fluorescent dyes with reactive groups, such as isothiocyanates, succinimidyl esters, maleimides,and carbodiimides. Reagents for this purpose can be found here, for example:
https://www.thermofisher.com/us/en/home/references/molecular-probes-the-handbook.html
After the labeling reaction, the labeled compound will have to be purified by chromatography.
It is essential to consider how the attachment of the fluorescent dye will affect the behavior of the compound, since fluorescent dyes are large organic compounds.
Dear Murad Alahdal
Fluorescent tagging or labeling, in general, employs a reactive derivative of a fluorescent molecule known as a fluorophore. The fluorophore binds to a specific region or functional group on the target molecule and can be chemically or biologically attached.
Have a look: https://promocell.com/cell-culture-basics/fluorescent-labeling/
Kind Regards
Qamar Ul Islam
Qamar Ul Islam Thank you very much sir. Your answer is very helpful to me, appreciate it.
Of course another consideration is what type of intrument you will be using. I presume a microscope but will you be using one or two photon excitation? Do your cells have significant autofluorescence to avoid? Probably you should use one of the newer photostable fluorophores.
David M Jameson Thank you very much, I definitely consider all of these issues. Thank you, sir.
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