Dear all,
Can anyone suggest me a dye for staining (living or fixed) of ALL membranes (also GolgI/ER etc.) in mammalian cells (adherent cells).
Thank you very much!
PS.: I already tried PKH67 and FM4-64FX which seem to only stain the plasma membrane.
You can try to use labelled Wheat germ agglutinin (WGA, eg- FITC Conjugated Lectin from Triticum vulgaris, L4895 SIGMA). It will be one of the cheapest option for you to give it a try.
WGA detects some of the glycoproteins mostly present on the plasma membrane. It can be used to stain both live and fixed cell. Though, WGA is mostly recommended for cell membrane, we have seen it to stain Golgi components along with plasma membrane. We have not seen any clear staining of ER. Although we have not explored that part as our aim was to stain cell membrane glycoproteins. You can try troubleshooting with it to check whether it works for your need, for example try after fixing and mildly permeabilizing the cell which will allow better internalization of WGA while keeping the cell membrane integrity. WGA will also stain any structures in the cell with high glycoprotein components (especially N-acetyl-glucosaminyl residues and its oligomers).
You can also use FM dyes, such as FM4-64, FM1-43 or FM5-95.
They are highly hydrophobic and thus have a very high affinity for membranes.
It is rather used for live staining, but it works well on fixed cells/tissues.
For concentrations read "FM-dyes as experimental probes for dissecting vesicle trafficking in living plant cells"
Article FM-dyes as experimental probes for dissecting vesicle traffi...
Lipophilic cyanine dyes, like DiI C18 (product D282 from Thermo Fisher) quickly intercalates through all cell membranes and would be a good choice. Be aware that these, as well as the FM dyes, will be lost if the cells are permeabilized in any way. (the PKH kits from Sigma use these dyes, but they also have some sort of sugar included, which changes the dye localization slightly for cell proliferation uses).
Amlan mentioned WGA. WGA and other lectins have been successfully used for labeling ER and Golgi in fixed and permeabilized cells, but the distribution is very cell-type selective, and it won't label all membranes.
Dear all,
thank you for all your answers. After testing several dyes, CellTrace™ BODIPY® TR Methyl Ester from ThermoFisher worked absolutely great (red fluorescence) for intracellular membranes (Golgi, all kind of vesicles, ER, nuclear membrane, mitochondria). I checked the stainings with a confocal microscope, I think a normal eppifluorescence would is not sufficient to resolve all the membrane structure (at least not with our setup).
Best, Jeff
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