If i have understood correctly, i think your interest is to show host cells that have internalized viral particles. DAPI (nuclear staining) of permeabilized cells will help you demarcate host cells. 

hm, interesting. theoretically, i first fix by FA, than stain for cell membrane by a lectin, than fix again by a crosslinker, next permeabilize and stain for virus. by this way, you can mark and fix the first signal on membrane, than you can label your intracellular target... 

usually the cell membrane markers does not allow the permeabilization, or you will see some internal signal too. that is why you should do in two step the whole procedure, and fix the first signal to ensure it remains on the membrane. One can give a simplier answer probably. i assume you want to stain only the membrane from some reason.

https://www.thermofisher.com/hu/en/home/life-science/cell-analysis/cell-structure/plasma-membrane.html

I am not sure if I understood the point of the exp, but if you just need to show where is the cell membrane, then you can preincubate cells on ice with any antibody against plasma membrane exposed protein/receptor on your cells (this depends on what kind of cells you have) and then fix immediately (not to get your Ab to be internalized). After this you simply permeabilize and stain with secondary antibodies. If you do not want to put your cells on ice, fixation with 4% PFA would work, but you need to have a fresh one each time, since once it gets old, you get some methanol in the solution which leads to cell permeabilization. So in my hands labelling on ice with primary antibodies worked best. If you need a protocol, check some articles where people do Flow cytometry on live cells :) Then they have cells in suspension, but the same can be done on attached cells with no problem. Good luck!

Hi Fatima,

You can use Phalloidine as a marker for the actin network beneath the membrane. 

Regards,

Ekta

Hi Fatima,

I suggest the CellMask orange fluorophore. Nice paper using this fluorophore, and performing real-time confocal fluorescence microscopy of ultrasound-mediated sonopermeabilization: Hu et al. 2013, Membrane perforation and recovery dynamics in microbubble-mediated sonoporation, Ultrasound in Medicine and Biology.

Good luck,

Best regards

Marc

Hi Fatima,, I would suggest the alpha-1 subunit of Na/K-ATPase which is a membrane located protein. The antibody from Upstate (Clone 464.6) is wonderful. Hope this will help. Good luck. Jiang

you can use WGA-Alexa fluor 594 (https://www.thermofisher.com/order/catalog/product/W11262), I used it to label the plasma membrane of neurons incubated with fluorescent exosomes, I was following their endocytosis. Sounds like you want to do more or less the same with your viral particles ^^ here is the paper we published http://www.ncbi.nlm.nih.gov/pubmed/25398455

Good luck with your experiments,

Mathilde

To trace the membrane in general you can try the membrane dye octadecylrhodamine (R18).

https://www.thermofisher.com/order/catalog/product/O246