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On Fri, 15 Nov 2002, Alison North wrote:

> 6)  ... My current PE
> system can either do GFP, Rhod and Cy5 together, OR CFP and YFP together,
>
> 7)  ... My PE system consists of
> a Zeiss microscope, the PE head and lasers etc., and then a load of filter
> wheels, cameras, software etc. from Universal Imaging,

Dear Alison,

Thank you very much for your nice comparison! Just one question: Have you
managed to automate filter swicthing on your PE system using filter
wheels? Where is the filter wheel situated (in your optical path)?

We have also got a Yokogawa CSU-10 system (which PE calls Ultraview)
and have struggled with its automation some time ago. Without success.
The confocal head contains a slider with 3 positions for 25mm exc. filters
(e.g. GFP+Rhod+Cy5) but it is very hard to replace it with a computer
driven filter wheel (without drilling into the confocal head).

Putting the filter wheel in front of the laser seems to cause severe
alignment problems (filter surfaces will never be parallel to each other
and will cause the light from the multiple-wavelength multi-mode laser
miss the 4 micron hole in the optical fiber for certain filters).

Another possibility is to put an AOTF between the laser and the conf. head
(fiber input and fiber output) but AOTF is very light inefficient (some
30-40% light throughput according to the specifications).

Thanks in advance for advice,
best regards,
Michal

P.S. Which laser (what laser power) do you use with your PE for GFP/Rhod/Cy5?
     We purchased a Melles-Griot one (Ar/Kr 35 LTL 835) with 10/10/15 mW
     for 488/568/647 nm, respectively (the strongest multiple-wavelength
     multi-mode laser from Melles-Griot) but find it quite week for most
     applications. The other possibility we found was Coherent Innova 70C
     (2.5W) but nothing in between these two extremes.

     Experience and ideas of other listers are also welcome.

~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~
Michal Kozubek, Ph.D.
Laboratory of Optical Microscopy, Faculty of Informatics,
Masaryk University, Botanicka 68a, CZ-60200, Brno, Czech Republic
Tel/Fax/Ans: +420-5-41512467   E-mail: [log in to unmask]
Internet home page: http://www.fi.muni.cz/~kozubek