http://confocal-microscopy-list.275.s1.nabble.com/Live-cell-imaging-experiences-please-tp4285373p4292834.html
My experience with a Nikon A1 is similar to what Neeraj described. We almost never see drift in focus or XY during long time-lapse imaging of many (>20) independent XY positions, and when we do it is invariably a software bug rather than a limitation of the hardware. Elements software also has a convenient function for 'tiling' several XY points into one larger image.
The 'optimize' function in Elements makes a decent go of minimizing travel between points, but it optimizes for a single time point (e.g., the last point is usually the most distant from the starting position). That sometimes causes the focus control to miss on the return due to the long travel. I find it quite easy to modify the order by hand to eliminate any large distance gaps.
We have never had a problem managing files up to several GB.
Based on my experience with other microscopes, 'stability' during long-term imaging usually entails focus drift due to thermal expansion/contraction and/or vibration. With its current hardware Nikon simply does not have that problem. I cannot speak for other contemporary scopes, but my success rate for time lapse > 4 h. has gone from ~20% on a dated, non-motorized system to almost 100% using the A1. The only reason we lose a movie is rare software malfunctions (the Vista edition of Elements has benefitted from a year of bug fixes).
I believe that Nikon's PFS hardware uses a unique TIRF-like infrared mechanism to maintain a constant distance between objective and coverslip. This allows a refresh rate in the milliseconds and it does not depend on detecting fluorescence from the sample. PFS will stay locked even if the software crashes and the computer has to reboot. Elements even remembers XY positions entered just before a software crash, which is nice.
That said, if product demos are an option then I recommend that you try your experiment on several systems to see which best matches your particular needs.
> Hi Zoltan,
>
> I have done long term time lapse (24 hour) experiments looking at multiple stage points in 3 different channels. We have a Nikon TiE with a motorized stage, perfect focus system and a Sutter Lambda 10-3 mechanical shutter. Its fairly easy to setup multidimensional experiments in the Nikon elements software, you can setup pretty complicated configurations which can be saved and reloaded. Using the perfect focus you can set different Z offsets for different stage points, there is also a function which automatically optimizes the stage travel between the stage points.
>
>
> As always, no commercial interests just a satisfied customer.
>
>
> Neeraj V. Gohad, Ph.D.
> Postdoctoral Fellow
> Okeanos Research Group
> Department of Biological Sciences
> 132 Long Hall
> Clemson University
> Clemson,SC-29634
>
> Please note my new email address:
[hidden email]
>
>
> -----Original Message-----
> From: Confocal Microscopy List [mailto:
[hidden email]] On Behalf Of Christian Götze
> Sent: Tuesday, January 12, 2010 5:36 AM
> To:
[hidden email]
> Subject: Re: Live cell imaging experiences, please
>
> Commercial response!
>
> Hi Zoltan,
>
> exciting application! Please correct me, if I'm wrong but one of these
> experiments would result in about 12.5 GByte of data (at 512x512 Pixels
> per plane, 8 Bit intensity, 18 Planes per stack, 8 stacks per time
> point, 1800 time points). What kind of software do you plan to use?
>
> If you don't have a suitable candidate already, perhaps you should have
> a look on our arivis Browser software. It is specialized in handling
> very large experiments and could be the right solution for you.
>
> (
http://www.arivis.com/en/Produkte/arivis-Browser)
>
> If you are interested or there are questions please don't hestitate to
> contact me.
>
> Christian
>
> Zoltan Cseresnyes schrieb:
>> Dear All,
>>
>> We are planning to run a few tests, where we follow cell development
>> in 8-9 XY positions, running 45 um Z stacks (step size 2.5 um), 2.5
>> min time interval, for a total of 14 hours at room temp, using GFP and
>> DsRed settings. We had lots of success with the Leica SP5 system for
>> the past 2 years. We are wondering if this community has more input
>> about how other systems would perfom here? We are most interested in
>> Olympus FV1000 and Nikon A1. Our main concerns are the simplicity of
>> the setting up procedure, the stability of the system during the long
>> scans, and the handling of the large data files.
>> All inputs will be much appreciated! Happy New Year to all! With
>> best wishes,
>>
>> Zoltan
>>
>
> --
> Christian Götze
>
> arivis - Multiple Image Tools GmbH
> Kröpeliner Straße 54, D-18055 Rostock
>
> Tel : +49 381 461393 11
> Fax : +49 381 46139399
> Mail:
[hidden email]
> Web :
http://www.arivis.com>
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> Geschäftsführung: Raik Madla, Christian Goetze