Greetings Howard and Radha: Sometimes simple works well. I have used the BD/Pharmingen CBA kit quite often. Although the selection of cytokines and chemokines are determined by the specific kit [although BD/Pharmingen are releasing custom kits], the sensitivity is very good, the kit results are very reproducible, it is easy to use and the kit includes a set of standards covering the range of analyte expression. The software is familiar to most operators [Cell Quest and Microsoft Excel] and acquisition and subsequent analysis is straightforward; being downloaded into Excel. BD/Pharmingen has done a nice job on working out many of the variables which the investigator would otherwise have to perform. This kit will be even better as more custom kits are available. Just my two cents. PS-This is my opinion and not reflecting any BD/Pharmingen ties or relationships. Hope this helps and all my best regards, -Rich Richard F. Konz, Jr. Scientist Manager, Flow and Image Cytometry Facility Wyeth 200 Cambridge Park Drive Cambridge, MA 02140 USA Office: 617-665-5522 Lab: 617-665-5551 rkonz@wyeth.com >>> Howard Shapiro <hms@shapirolab.com> 09/15/02 01:28PM >>> Radha Dutta-Roy wrote: > Have any of you who regularly use CBA kits(by >B.D.Pharmingen) ever tried using kits made by other companies(like >Upstate's BeadLyte Cytokine Detection Kit) on the flow machine.The >principles are the same so why would you for example have to use a luminex >machine to run Upstate's kit and not try running it not the Flow >Cytometer.(legally I am sure Upstate would say no you can't run it on the >flow machine blah blah!!!and that they can't guarantee results >blah!blah!)Has anyone out there thought about it or tried it???Could you >please share your thoughts with me, I would really appreciate it.THANKS! The Luminex machine uses a 532 nm green laser to excite reporter molecules (phycoerythrin, Cy3, etc.) and a red laser to excite red and near infrared fluorescence from the dyes used to color code the beads. Kits designed for use with this machine could theoretically - and I emphasize theoretically - be made to work on a flow cytometer with 488 nm and red (633 nm HeNe or 635 nm diode) lasers if two fluorescence channels were available on the red laser, but sensitivity would not be as good because 532 nm excites PE better than 488 nm, while 488 nm excites autofluorescence and background fluorescence better than 532 nm. And one would need all the software to decipher which bead was which; the Luminex software deals with as many as 100 different shades of beads, while the current B-D and Bangs Labs bead sets have many fewer shades and use simpler software. Luminex originally made a bead set using fluorescein as a reporter and red and orange dyes for bead color coding; the beads could be run on a FACScan using Luminex's add-on computer system and software to do the analysis. However, Luminex isn't making those beads, or the add-on hardware and software, anymore, for obvious reasons. If you want to do assays using the new beads, it will be far easier to use Luminex's instrument; even if you manage to build or modify your own box and write your own software to get equivalent performance (not a trivial undertaking), you might be infringing on one or more Luminex patents, and, if you intend to use the assay for clinical purposes, you might also have to worry about FDA and CLIA issues. In the interest of full disclosure, I have worked and continue to work with Luminex on instrument design, among other things. However, I wouldn't have written anything different in answer to the question if I had no ties to the company. -Howard
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