[Cytometry] Sorting with PerCP conjugates
jl7fj at virginia.edu
Mon May 9 11:47:31 EDT 2011
We regularly sort PerCP-Cy5.5 with a 200mw laser without any problem. The excitation of the PerCP portion seems to still allow the energy transfer to the Cy5.5. molecule. Our users prefer this over PE-CY5.5 because there is significantly less compensation out of the PE channel. We find that laser powers at 50 mw and below are fine for PerCP. So don't abandon PerCP Cy5.5 on your Vantage, it will work.
On Mon, 09 May 2011 16:20:20 +0100
"Rosemary Clarke" <r.z.clarke at dundee.ac.uk> wrote:
>We currently have a BD Vantage/DIVA cell sorter and routinely analyse antibody stained cells with an Enterprise laser 488nm at 250mW. (Lower than this and we don't get sufficient UV light out). Because of the issues associated with using PerCP (poor signal strength, photobleaching, stability etc) we have always avoided using it for cell sorting, along with it's conjugates (such as PerCP/Cy5.5) and opted for PE/Cy5.5 instead. We are increasingly finding that manufacturers are no longer providing PE/Cy5.5 conjugates, opting for the PerCP alternative. Obviously this conjugate has huge advantages for analysis, but not for sorting.
>We are currently putting in a proposal to purchase an Aria III sorter, which as standard will have a 20mW 488nm laser. My question is, does the lower laser power and optical set up of the Aria mean that it will be possible to use PerCP and it's conjugates for sorting? My thinking is that we get a great signal on out LSR Fortessa so why not on the sorter?
>A very simple question with a simple answer, I hope.
>Dr Rosie Clarke
>Division of Cell Biology and Immunology
>University of Dundee
>Tel: (01382) 384222
>E-mail: r.z.clarke at dundee.ac.uk
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Joanne Lannigan, MS
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University of Virginia
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