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Michael Hooker Microscopy Facility (MHMF.ORG) |
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Step Size recommendations for Z-series | |||||||
on the LeicaSP2 confocal |
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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 2001-2014 Dr. M. Chua, Program in Molecular Biology & Biotechnology, School of Medicine, University of North Carolina, Chapel Hill, NC 27599 |
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Questions/comments/problems: Michael Chua |
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Last Updated:
2014-07-23 |