Flow Cytometry Technical Question >>>>

Discussion in 'Physician Scientists' started by GWD, May 13, 2008.

  1. GWD

    GWD
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    Does anyone know if/how to do post data acquisition compensation for flow cytometry?

    I'm a relative n00b when it comes to this machine/software.

    I'm using a Beckman Coulture FC500 flow cytometer running on CXP software.

    Basically I ran my samples and acquired my data, but now I realize that I needed to use some heavy compensation to normalize the data.

    So, does anyone know if it is possible to do post acquisition compensation, and if so... how?

    (I'm not calling tech support because our service agreement is up and I don't want to remind them to come pick up the machine)
     
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  3. Tiger1

    Tiger1 UIUC-VMSP (DVM/PhD) Y2
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    I used BD FACSCalibur for analysis and FACSAria for sorting extensively. I analyze via FlowJo. I know that you can NOT compensate post-acquisition on FlowJo because it is a mathematical formula applied to the data during calibration prior to acquisition.

    However, if you ran negative control samples with your actual experimental set for EACH of the colors that you labeled, you can gate the negative set and anything "significant" above or the right of the gate could be positive. You will have to interprete the data critically.
     
  4. GWD

    GWD
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    Thanks for the info... I don't think it's applicable to the software I am using

    "supposedly" you can do post-acquisition compensation on this software, just nobody in the lab knows how

    must be nice running a FACSAria though *drools*
     
  5. MedRower

    MedRower Member
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    I'm curious, is there someone else at your institution who uses the software who you might ask?
     
  6. GWD

    GWD
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    No. If there was, don't you think I would have asked them already?

    Of course we have a CORE facility here, but they run different machines with different software.

    This is something akin to a personal machine which pretty much just our lab and 1 or 2 other individuals ever use, which is why the department voted against extending the pricey service agreement. Everybody seems to agree that it can be done, but nobody quite knows how.

    It's no big deal.. it's probably only 3-4 hours of work to re-do, I just thought somebody might be familiar with how it was done in case this problem ever comes up in the future.
     
  7. MedRower

    MedRower Member
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    Shrug. Of course man, but you know how un-straightforward things can be when there are people in the building who have experience with a piece of equipment that you aren't aware of until you ask around.
     

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