HELP: Performance spinning disk Andromeda/TILL

[offterdi1]

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Dear all,
We have a brand new iMIC/TILL which shows serious performance problems if
used in the spinning disk mode ("Andromeda", not exactly spinninhg disk but
rather spinning mirrors). Although we tested a very similar system last year
using the same type of CCD cam etc.. we now experience a very dissappointing
light sensitivity and a low signal/noise ratio compared to last year. Can
anyone point me in a useful direction? The Till people have only been of
very limited help and more or less argue that everything is perfect.
What are the likely error sources? Camera chip to chip variation (= same
model!) Laser alignment, optical fibres, lenses in the light path????

I think i will need to prove what is wrong exactly in order to get proper
support fromt the company.


Best
Martin

[Csúcs Gábor]

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Dear Martin,

I'm really wondering the insufficient answer from TILL as so far we always
experienced them as a very helpful people who really try to find a
solution to your problem. Of course, right now this doesn't help you too
much.
The first obvious thing I'd test is the laser power you have on the system
(which is of course strongly dependent on the alignment). Your should also
check whether the filters you are using are the same as used during the
demo. Of course there are variations also on the camera/objective side as
well but according to you the problems occur only when using the confocal
unit. A very obvious thing would be to do to test your systems once more
with the demo system you have used last year and compare the results. If
you really see huge differences - there is something to show to TILL. When
we demoed the Andromeda we also found it very convincing, so I don't
believe that your problems have to do with the technology itself.

Greetings     Gabor



On 10/31/12 8:56 AM, "Martin Offterdinger"
<[hidden email]> wrote:

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>
>Dear all,
>We have a brand new iMIC/TILL which shows serious performance problems if
>used in the spinning disk mode ("Andromeda", not exactly spinninhg disk
>but
>rather spinning mirrors). Although we tested a very similar system last
>year
>using the same type of CCD cam etc.. we now experience a very
>dissappointing
>light sensitivity and a low signal/noise ratio compared to last year. Can
>anyone point me in a useful direction? The Till people have only been of
>very limited help and more or less argue that everything is perfect.
>What are the likely error sources? Camera chip to chip variation (= same
>model!) Laser alignment, optical fibres, lenses in the light path????
>
>I think i will need to prove what is wrong exactly in order to get proper
>support fromt the company.
>
>
>Best
>Martin

[Josef Gotzmann]

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Hi Martin,

I can only back Gabor's comments.
We have demoed the Andromeda system when it was brand-new and in two demo
weeks we experienced similar problems and TILL being stumped with pulling
out enough power of the system. Although using the mirrors promises a gain
of intensity we did not see this increase compared to double-Nipkov systems.
Laser alignment was very critical at the Andromeda when we tested it as was
usage of appropriate filters - I would also suggest to measure laser power -
you should have data from installation to compare.
best,
Josef


 
On Wed, 31 Oct 2012 02:56:35 -0500, Martin Offterdinger
<[hidden email]> wrote:

>*****
>To join, leave or search the confocal microscopy listserv, go to:
>http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
>*****
>
>Dear all,
>We have a brand new iMIC/TILL which shows serious performance problems if
>used in the spinning disk mode ("Andromeda", not exactly spinninhg disk but
>rather spinning mirrors). Although we tested a very similar system last year
>using the same type of CCD cam etc.. we now experience a very dissappointing
>light sensitivity and a low signal/noise ratio compared to last year. Can
>anyone point me in a useful direction? The Till people have only been of
>very limited help and more or less argue that everything is perfect.
>What are the likely error sources? Camera chip to chip variation (= same
>model!) Laser alignment, optical fibres, lenses in the light path????
>
>I think i will need to prove what is wrong exactly in order to get proper
>support fromt the company.
>
>
>Best
>Martin

[offterdi1]

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Dear Josef and Gabor
Thanks a lot for your input. In our case we very also extremely satisfied
with the demo last year, and also the Till staff and therefore decided to
buy it. But now we are very dissatisfied with our new system and also their
support and I start thinking that it was a bad investment. Anyhow...

I don't think that laser power per se is our problem, we rather have more
power than the demosystem and also filters etc are the same (Quad). I am
rather hunting for some alignment errors, fibres etc since we do not get
good images, high bleaching rates, basically it can't be used in the same
experimental setup which was excellent last year. (FRAP, Andromeda
combination with a low GFP expressing sample)...


Best
Martin

[Csúcs Gábor]

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Dear Martin,

The question is where do you measure the laser output: at the end of the
fiber or in the sample plane? Of course it can well be that there is a
problem with the alignment of the Andromeda itself (misaligned disk?) But
naturally this can find out only with the help of TILL. You could also try
to image fluorescent beads and look whether their shape looks normal.
Spherical aberrations can also lead to very weak signals and high
bleaching rates.

Greetings    Gabor



On 10/31/12 12:11 PM, "Martin Offterdinger"
<[hidden email]> wrote:

>*****
>To join, leave or search the confocal microscopy listserv, go to:
>http://lists.umn.edu/cgi-bin/wa?A0=confocalmicroscopy
>*****
>
>Dear Josef and Gabor
>Thanks a lot for your input. In our case we very also extremely satisfied
>with the demo last year, and also the Till staff and therefore decided to
>buy it. But now we are very dissatisfied with our new system and also
>their
>support and I start thinking that it was a bad investment. Anyhow...
>
>I don't think that laser power per se is our problem, we rather have more
>power than the demosystem and also filters etc are the same (Quad). I am
>rather hunting for some alignment errors, fibres etc since we do not get
>good images, high bleaching rates, basically it can't be used in the same
>experimental setup which was excellent last year. (FRAP, Andromeda
>combination with a low GFP expressing sample)...
>
>
>Best
>Martin