The exact amount of dye needed will depend on the quantum yield, the quality of your fluorescence microscope and so on. Generally you're looking at around 1-3 mol%. For dyes like Texas Red (I usually used Texas Red DHPE) you can get away with 0.25 - 0.5 mol%, for NBD I'd recommend higher concentration to get good signal to noise, as it photobleaches really easily (I've had good experiences with concentrations around 2 mol% but sometimes had to go even higher). Dyes from the Atto family can now also be bought attached to lipids, they are so photo stable that it can be hard to bleach them at all, but you will need even less dye than for Texas Red, if you're using Atto 590 something like 0.1 mol% can still be fine.

Thanks Johannes, I remember, during my training on confocal fluoresence microscope two years ago, instructor used GFP (green), YFP (yellow) and RFP (red) to prepare a sample. Can I use anyone of the these dyes in lipids? Some literature mention that these dyes are for proteins.  

i prepared lipid bilayer membrane using vesicles with lipids: DPPE and DPPS with ratio 2:1. How can i attach dye with it? Which type ofdye is most suitable? I will estimate diffusion coefficient of bilayer membrane deposited on solid state nanopore biosensor. 

GFP, YFP and so on are fluorescent proteins. You don't want to use them for the characterisation of your membrane because these aren't membrane proteins and are soluble in water. Therefore you won't be probing your membrane but either the solution in the vesicles or the surrounding medium. If you want to probe the membrane itself you want to have dyes attached to lipid molecules. The lipids will then be in your membrane and will behave similar (or identical) to the lipids in the membrane.

Dye labelled lipids are available commercially. For example Texas Red DHPE can be bought here https://www.lifetechnologies.com/order/catalog/product/T1395MP (or from many other sources, just search for the cheapest option for you) and you can find a variety of labelled lipids at avanti http://avantilipids.com/index.php?option=com_content&view=article&id=261&Itemid=213

Sadly labelled lipids are much more expensive than lipids without a dye, but keep in mind that you only need around 1 mol%

Thanks Johannes. In literature, I noticed that Biotin DHPE, Fluorescein DHPE,  Rhodamine-DHPE and Texas Red-DHPE are the potential dyes for lipids. Why Texas Red DHPE is expensive as compared to others. I found the least expensive on this web link:

https://biotium.com/product/texas-redtm-dhpe-n-texas-red-sulfonyl-12-dihexadecanoyl-snglycero-3-phosphoethanolamine-triethylammonium-salt/

Also do you have any procedure/protocols for attaching TR-DHPE with bilayer membrane. I prepared LBM with final concentration 20-25mM by suspended lipids (10mg/mL) in n-decane. 

You can use any of the dyes you identified, they should all work for your purposes. I'm not sure why Texas  Red is expensive compared to the others, I like using it for its photostability, but you might not need this.

The incorporation of the dyes into the membrane is a rather simple process. They are already attached to a lipid so you can just add the dye when you are preparing the lipid mixtures. Instead of just using 2:1 DPPE:DPPS you can simply prepare something like 100:50:1 DPPE:DPPS:[Dye] DHPE, depending on the dye you choose you might want slightly more dye or might get away with using less. The lipids will then be in your membrane and the dye molecules will stay attached to the lipids, you should be able to easily see your membrane then.

Thanks Johannnes. I have to say a big thanks and appreciate your discussion. I will inbox you my results once I am done for more discussion. Send me your email. 

Any website you recommend for buying GFP. 

Hi guys!, very interesting topic!. I have the next question: I want to use the DHPE not to study the plasma membrane but to label it and track my cells. I want to see if they are exchanging membrane each other. Can you suggest to me a good protocol for use this dye?. I read that you have to resuspend in clorophorm but nothing else about how to use that suspension. Thanks for your time!