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Step Size recommendations for Z-series

on the LeicaSP2 confocal

 
For multi-color images, use the optimal step size for the fluorophore with the shortest wavelength
 
  Pinhole 1 Airy Unit
  Excitation Laser UV 488 561 633
  ~ Em. Wavelength 460 520 600 670
  Typical fluorophore DAPI FITC Rhod Cy5
Objective

(mag / NA)

10X 0.3 6.7 7.6 8.8 9.8
16X oil 0.5 2.4 2.7 3.1 3.5
20X 0.7 1.2 1.3 1.5 1.7
40X oil 1.25 0.31 0.35 0.40 0.44
63X oil 1.4 0.21 0.24 0.28 0.31

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Copyright 2001-2015  Dr. M. Chua, School of Medicine, University of North Carolina, Chapel Hill, NC 27599
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Last Updated: 2014-07-24
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