How to detect receptor on the cell surface? - using the less viral protein possible (Oct/10/2005 )
Hi,
I have recombinant viral envelope protein, and I want to detect with this a receptor on the cell surface.
I have all I need - cells, viral protein ( but unfortunately not so much), which is biotinylated, and conjugate: streptavidin - fluorescent molecule.
My question is - what cell volume , the minimun, can I utilise to perform the recognition procedure, and to see the clear result under fluorescent microscope?
I need you suggest me the concrete plastic dish ( 96 wells, 24 wells, 6 wells, which diametre?)
Can we observe the reaction with fluorescence microscope only with drop on hemocytometre, or it is possible to observe directly culture plate?
My other question - what the minimal volume of fluorescence -marked celles could be counted with standard FACS ?
I have recombinant viral envelope protein, and I want to detect with this a receptor on the cell surface.
I have all I need - cells, viral protein ( but unfortunately not so much), which is biotinylated, and conjugate: streptavidin - fluorescent molecule.
My question is - what cell volume , the minimun, can I utilise to perform the recognition procedure, and to see the clear result under fluorescent microscope?
I need you suggest me the concrete plastic dish ( 96 wells, 24 wells, 6 wells, which diametre?)
Can we observe the reaction with fluorescence microscope only with drop on hemocytometre, or it is possible to observe directly culture plate?
My other question - what the minimal volume of fluorescence -marked celles could be counted with standard FACS ?
You really only need enough to put on a slide and look at them so a couple of hundred cells would be enough and a 96 well plenty. Using FACS 10000 cells are generally counted but to comfortably have enough so the cytometer won't run dry etc I like to 10^5.