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  • fluorescence microscope widefield laser illumination

    From Fitzgerald@21:1/5 to All on Fri Jul 15 10:47:27 2016
    I would like to use laser illumination for a widefield fluorescence
    microscope. I am wondering how to handle the polarization and/or
    coherence effects.

    I have read that it is better to have circular polarization on the
    sample, to avoid any preferential excitation of fluorophores with a
    specific orientation. How much of this is true and how much is superstition?

    Secondly I am wondering how to combat laser speckle to get uniform
    illumination of the sample. I might focus the laser in the back focal
    plane of the objective and then move the spot around in the BFP by
    adjusting the tip/tilt of the mirror. There are probably other ways.
    What is generally used for this?

    Thanks in advance

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  • From RustyHinge@21:1/5 to Fitzgerald on Wed Sep 21 13:04:18 2016
    On 15/07/16 09:47, Fitzgerald wrote:
    I would like to use laser illumination for a widefield fluorescence microscope. I am wondering how to handle the polarization and/or
    coherence effects.

    I have read that it is better to have circular polarization on the
    sample, to avoid any preferential excitation of fluorophores with a
    specific orientation. How much of this is true and how much is
    superstition?

    Secondly I am wondering how to combat laser speckle to get uniform illumination of the sample. I might focus the laser in the back focal
    plane of the objective and then move the spot around in the BFP by
    adjusting the tip/tilt of the mirror. There are probably other ways.
    What is generally used for this?

    Thanks in advance

    I find Kohler illumination does the biz.

    --
    Rusty Hinge
    To err is human. To really foul things up requires a computer and the BOFH.

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    * Origin: fsxNet Usenet Gateway (21:1/5)