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Determine optical slice thickness in a CLSM

  1. Dec 12, 2008 #1
    I am trying to figure out the optical slice thickness on a confocal laser scanning microscope with the following:

    Lambda exitation: 514
    Lambda emmision: 548nm
    Pinhole: 2 AU / 222,9 um
    NA: 1,2
    n (refraction index): 1,33 (W)

    I have tried a few equations, but they give quite different results. One of the equations I have used is in this article from Zeiss in the following link:
    http://www.zeiss.com/C1256D18002CC306/0/F99A7F3E8944EEE3C1256E5C0045F68B/$file/45-0029_e.pdf
    (page 11, equation 4)



    I am going to use this in my Master thesis due soon. Can someone please help me determine the correct way of measuring this?
    I must stress that I am a biologist so "be gentle" with the complex formulas :wink:
     
  2. jcsd
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