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  • mh123
    Member
    • Feb 2008
    • 3

    #1

    Focusing problem with new flowcell on GAII

    I have focusing problems with 1.4mm FC. Sometimes a couple of lanes will be totally off focus while the rest of the lanes seems to be within the 15000nm range; Sometimes one end is +10,000nm and the other end gives -10,000nm and this makes it impossible to set a good "Z" value for focusing across the full FC. I think part of the problem is caused by the container FC was stored/shipped because often FC got stuck in it and this may have squeezed and forced the FC to change shape since it's a very thin sandwich. Anybody having similar issues in the field? Any solutions or suggestions? Thanks!
  • monad
    Member
    • May 2008
    • 40

    #2
    Originally posted by mh123 View Post
    ........and this makes it impossible to set a good "Z" value for focusing across the full FC.........
    Not every FC is manufactured perfectly....generally speacking, as long as you have a good focus in the middle of FC, lane 4-1,2 or 5-1,2, you may be fine.
    If way off, it seems your case, chances are plate that holds your FC in your GAII may be off-leveled. Illumina engineer has a tool to check and adjust the level. top left, right and bottom left and right should be leveled for even focusing. This off-level may cause increase error rate as it will affect imaging quality. You may want to check with FAS and/or service engineer.
    Last edited by monad; 11-21-2008, 09:31 AM.

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