Posted by
leoncio vergara on
URL: http://confocal-microscopy-list.275.s1.nabble.com/stage-motorization-tp4455148p4460416.html
I think you are absolutely right.... despite the stability issues, having a motorized stage in your system really add to the capabilities and the possibility to do multi-locations is a real advantage.
One has to remember however that for multi-location work you absolutely need a good fast autofocus system, this adds to the cost if budget is concern, but a the stage alone would not be enough... even if the company tells you that their microscopes are extremely stable.... .
________________________________________
From: Confocal Microscopy List [
[hidden email]] On Behalf Of Keith Morris [
[hidden email]]
Sent: Tuesday, January 26, 2010 4:26 AM
To:
[hidden email]
Subject: Re: stage motorization
I agree with all the previous comments, but from our point of view:
Personally I'd always buy a motorized stage, whether time-lapse is involved
or not - although this does assume it's on a more expensive fully motorized
microscope [filter wheels/focus]. Back in the days when I bought my own
microscopes they always came with one. In recent places I have worked at,
the confocals have arrived before my time, and a manual stage has always
been fitting and XY drift during long time-lapse [say 78 hours] is an issue.
It's not such that the time lapse can't be done but it is irritating. I say
drift, but normally it's more a small jump or two that's clearly not cell
movement [no doubt tension stored in the screw rack somewhere releasing
itself or 'vibration/thermal expansion'].
We have a manual XY stage now on our otherwise motorized time-lapse Zeiss
510 Meta confocal and really miss the ability to go back to the same
location [say take one live cell photo every day at the same place], or to
time-lapse multiple wells, perhaps with different fluorochrome labels [and
get the reduced stage drift]. If a motorized stage had been specified in the
original quote we would got it for 'free' [if a confocal costs a £220,000 or
£230,000 it's not going to make much difference to whether the purchase goes
ahead, but trying to raise £20,000 after the confocal arrived will be a very
different matter. Ensure you have all the right time-lapse/3D/FRET/FRAP
modules you require factored into the quote as well [they are £5k each
afterwards], and the required phase contrast objectives for time lapse [air
Ph 20x at least, DIC isn't really good enough for transmission cell motility
time-lapse - we don't have a standard fluorescence/brightfield microscope
time-lapse system]. With multiple wells you might want to add a
non-immersion air hi-power objective as the oil won't easily move across.
A motorized stage will also allow a complete automated raster scan image of
a fixed slide [ideally via say Zeiss Axiovision and a separate CCD camera -
both ideal for transmission time-lapse], something else we really miss not
being to do. Likewise we could have automated metaphase finding, scoring and
relocation with our cytogenetics FISH workstation, but that has a manual
stage as well adding hours to our weekly routines [not 'confocal' I know,
but that missing motorized stage is still a pain].
We have just bought a large Zeiss XL3 incubator, TempControl & CO2 system
that protects the manual stage/objectives from drift somewhat during
time-lapse, but are now, as most users are asking for one, trying to raise
yet another £20k for the Zeiss motorized stage and associated control module
[plus another £5k for the FRET module], and it's probably going to take a
long while for the stage funding to appear.
The Zeiss motorized XY stage software module also allows us to have a
variable time-lapse: say 10 mins at one time interval and then the next 6
hours at a longer one [which our present 'physiology' module can't do].
Shame we never got it all with the microscope in the first place [and then
it probably would have attracted a large discount].
Good luck with the purchase anyway
Keith
---------------------------------------------------------------------------
Dr Keith J. Morris,
Molecular Cytogenetics and Microscopy Core,
Laboratory 00/069 and 00/070,
The Wellcome Trust Centre for Human Genetics,
Roosevelt Drive,
Oxford OX3 7BN,
United Kingdom.
Telephone: +44 (0)1865 287568
Email:
[hidden email]
Web-pages:
http://www.well.ox.ac.uk/molecular-cytogenetics-and-microscopy-----Original Message-----
From: Confocal Microscopy List [mailto:
[hidden email]] On
Behalf Of Craig Brideau
Sent: 25 January 2010 19:08
To:
[hidden email]
Subject: Re: stage motorization
A motorized stage will not necessarily be more stable than a
non-motorized stage. It mainly depends on the bearing system. The
motor just replaces your finger turning a shaft in most cases, so it
all really boils down to the mechanics of how the shaft is geared and
the bearing system the stage is resting on. That said, a motorized
stage with active correction built in, like 'Perfect Focus' will 'lock
on' to a given position and actively maintain it. More generally
speaking, if your room is fairly temperature stable, your microscope
is vibration free, and your ventilation system is not turbulent, you
should get decent stability from most good quality stages, either
manual or motorized. At the end of the day though, it will all
revolve around your specific case; how much movement can you tolerate
within what timeframe? You will need to verify that the specs of the
stage match your requirements. Even then, you will probably need to
test it out to be sure.
Craig
On Mon, Jan 25, 2010 at 9:56 AM, Charles Stevens <
[hidden email]>
wrote:
>
> Dear List,
>
> We're planning to set up an inverted confocal microscope for live cell
> imaging. I'd appreciate any opinions on using manual vs x-y motorized
stage.
> Is the motorized stage more stabile during long-term experiments? Does it
> facilitate/is it necessary for any experiments besides multi-point
> time-lapse imaging?
> Thanks for your comments!
>
> Charles
>
>