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September 2008

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From:
"Petrak, Lara J." <[log in to unmask]>
Reply To:
Confocal Microscopy List <[log in to unmask]>
Date:
Thu, 18 Sep 2008 10:41:15 -0400
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Hi Scott,

I agree with Gabor and Simon that having some sort of "auto focusing"
mechanism (whether it be a z-motor based software autofocus or a
laser-based perfect focus system) is extremely helpful, and sometimes
essential, when doing long-term timelapse imaging.  I find that even
with temperature and humidity control, z-drift is likely over a long
period of time and needs to be corrected for.  If you plan to image
fluorescence with live cells over time, imaging multiple planes at each
time point is not ideal because of the burdens of increased
photobleaching and phototoxicity.

I have used both software autofocus, which tends to work just fine, and
the Nikon Perfect Focus System on the TE2000E.  The PFS is easy to use
and works very well (and much quickly than traditional autofocus!).  If
you look into this, make sure that the objective lenses that you plan to
use are rated to work with the PFS (I think that many will work, but
there are some exceptions).

Best,
Lara

----------------------------------------------------------------------- 
Lara Petrak 
Microscopy Coordinator 
  
Nikon Imaging Center @ Harvard Medical School 
240 Longwood Avenue 
Building LHRRB Room 113 
Boston, MA 02115 
  
Systems Biology Microscopy Facility 
220 Longwood Avenue 
Goldenson Room 107 
Boston, MA 02115 
  
Phone: 617/432.3547 
Fax: 617/432.1144 
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