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    • CommentAuthoraadeal
    • CommentTimeNov 17th 2010 edited

    This protocol explains how to: Image Using a 100x Objective (Oil Immersion)

    Protocol revised: 11/17/10

    Protocol written by: Aaron Deal

    Note: When imaging a slide using a 100x objective, oil is required for a precise image.
    1. View the slide on a compound microscope. Begin on a lower powered objective and then move up to a dry 100x objective in order to find the desired, or potentially desired, image. (If the image is brightfield, see the Kohler illumination protocol for instructions on setting up Kohler illumination)
    2. Once the image is in view, rotate the nosepiece so that no objective is pointing towards the slide.
    3. Place 1-2 drops of objective lens oil (located on the shelves above the microscope) directly onto the slide where the objective will be located.

    4. Rotate the nosepiece back to the 100x objective, without passing any other objectives through the oil (the 100x objective lens should now be surrounded by the oil).
    5. The stage can still be moved, but a reapplication of oil may be required if the oil is spread too thin. It is best to move the stage as little as possible after applying oil.
    6. Once finished with the 100x objective, rotate the nosepiece so that there is no objective pointing toward the slide.
    7. Remove the slide and wipe with a kim wipe or any absorbent paper. DO NOT USE A KIM WIPE OR TISSUE ON THE OBJECTIVE LENS!
    8. Only use an objective lens wipe (found in packets on the shelves above the microscope) to clean the objective. Lowering the stage can give you more room to adequately maneuver in order to clean the lens.
    9. Place a lens wipe between your finger and the lens. Pull the paper without moving your finger so as to drag the lens wipe across the objective lens. Do not simply wipe in circles, this will only smear the oil, not remove it from the lens.
    10. If any other objective accidentally passed through the oil, clean the lens using the same technique described in the previous step.

    *If the objective still looks foggy or unclean for any reason, see the lens cleaning protocol for more options.*

    • CommentAuthorpulakhmani
    • CommentTimeDec 15th 2010
    Only use oil immersion techniques on objectives that were designed to be used with oil.
    • CommentAuthorelpitts
    • CommentTimeDec 16th 2010
    We usually use Type A Immersion Oil, but for our purposes either type of oil will likely work.