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    • CommentAuthorjuruble
    • CommentTimeJul 27th 2007 edited

    This protocol explains how to: align the phase on the Olympus fluorescent microscope.

    Protocol revised: 6/27/2007

    Protocol written by: Barbara Lom, Fiona Watson, Julie Ruble


    NOTE: While following this protocol, please reference the labeled diagram found below.  A full-sized version of this image is available here.


    1. Start with the 10X objective and make sure the Phase Selector Ring is on "Ph 1."  Focus on a slide.
    2. Take out the right eyepiece of the Olympus fluorescent microscope and insert the UT30 centering telescope (found bubble-wrapped in a plastic jar by one of the scopes).
    3. Push in and turn allen wrench knobs until both rings viewed through the telescope eyepiece align in the dark circle.  Make sure the allen wrenches are catching in the screw hole when you push them in to turn them.
    4. Repeat this with each objective and its corresponding Phase Selector Ring setting.  For the 40X objective, the ring should be on "Ph 2," and for a 100X objective, it should be set on "Ph 3."

    • CommentAuthorpulakhmani
    • CommentTimeDec 15th 2010
    Don't forget to put the telescopic lens back where you found it.
    • CommentAuthorsapyfrom
    • CommentTimeDec 15th 2010

    Be gentle with this process. These are small, fairly delicate parts and small adjustments are usually all that is necesary.

    • CommentAuthorroclemente
    • CommentTimeSep 29th 2011
    Phase alignment should be the first thing you check when you want to take a phase contrast picture.
    • CommentAuthormamccreery
    • CommentTimeSep 30th 2011
    Finding and focusing on an unstained specimen can be difficult if the microscope has not yet been properly aligned for phase contrast. It is often helpful to correctly Kohler and focus the microscope in brightfield, either on your specimen or another stained slide at the appropriate magnification, before switching to phase contrast.
    • CommentAuthorcpgruber
    • CommentTimeOct 1st 2011
    Remember the telescopic lens is like a focusing eyepiece, and it needs to be in focus before adjusting phase alignment.