Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal

Hi Ricardo,

Yes, adding 11 mm space under the objective lens for the piezo will 
affect the image on your 160 mm (171 mm) tube length microscope. If 
you are only interested in one plane, no need for the piezo. If you 
have not purchased the piezo, an external focus motor could be fitted 
to the focus knob if/when you do decide to acquire Z-series.

Yes, a high NA water immersion lens would be nice, but if you could 
use mounting medium that matches your immersion oil's refractive 
index (i.e. 1.515, same as glass), you would be fine. Of course your 
goal is to image live cells. You could boost the R.I. of your live 
cell culture medium using BSA, etc. to get at least close to index 
matching. Let us know if it makes a significant difference! Since 
your cells are probably within a couple of micrometers of the 
coverglass, even standard aqueous vs immersion oil refractive index 
difference will not be a killer - after all, plenty of researchers 
have published live cell microscopy with oil immersion lenses.

2D deconvolution is part of Fovea Pro 4.0, a set of Photoshop 
plugins. See the bottom of 
http://www.reindeergraphics.com/index.php?option=com_content&task=view&id=206&Itemid=152 
for example (astronomy). You can download an 3-week, fully 
functional, version of Fovea from the company at
http://www.reindeergraphics.com/index.php?option=com_content&task=view&id=43&Itemid=68

They also have a separate plugin, Focus Extender 1.0, that may be of 
use with Z-series 
http://www.reindeergraphics.com/index.php?option=com_content&task=view&id=29&Itemid=52


Best wishes.


Sincerely,


George


At 10:58 PM 3/22/2006, you wrote:
>Search the CONFOCAL archive at
>http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
>
>Hi all
>a question about deconvolution (again!!!!!)
>I am trying to put together a "home made" time lapse microscope (for gfp
>tag proteins and slow calcium signals) with deconvolution in mind,
>would something like this work?
>
>-a IM 35 zeiss inverted microscope (the 160 length tube!!) with a good
>63x/NA 1.3 oil objective (but no luck getting a water one so far)
>- a spot 2 (slider) ccd camera (diagnostic instruments)
>- a jena piezo device for the objective (any bet if the extra 11 mm will
>interfere with the optics?)..this is optional
>- a shutter for the mercury lamp and an optical excitation switch (DX-1000,
>Solameretech)
>- run by and old version of QED imaging software
>
>  the deconvolution would be made afterwards image acquisition. I don't plan
>to do any fancy 4D imaging, just one picture (one focal plane) over time,
>but most deconvolution free software (ImageJ and Image Track) I had found
>seem to be done for xyz stacks, I was able to trick the software supplying
>a psf made  of only one picture (not much of a psf...), but I am not sure
>if this is the rigth procedure, do you think it is posible to get good
>plain 2d deconvolution?  thanks....
>
>Ricardo Armisen
>
>SUNY at SB
>
>U. de Chile




George McNamara, Ph.D.
Division of Cancer Immunotherapeutics and Tumor Immunology
City of Hope National Medical Center
1500 E. Duarte Rd
Duarte, CA 91010
626-359-8111 ext 60035 office
818-547-6909                 home
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