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Dear All,
We experience quite visible axial thermal drift with the Prior H117
motorized stage under TIR illumination (ca. 10 nm per second) with the Nikon
100x N.A. 1.45 objective.
I have contacted Prior and I was told that they never measured thermal drift
for the H117 stage at that level - 100-200 nm. XY motors attached to the
bottom of the stage, thus the heat generated by the XY motors could also
contribute to the drift.
Nikon offers PFS or their Ti50 scope to solve the problem, a kind of
expensive solution for the Nikon's first generation, visibly
under-engineered TIR system.
I would greatly appreciate any positive feedback/suggestions.
Thank you,
Vitaly
NCI-Frederick,
301-846-6575
-----Original Message-----
From: Confocal Microscopy List [mailto:
[hidden email]] On
Behalf Of Cameron Nowell
Sent: Tuesday, February 10, 2009 1:00 AM
To:
[hidden email]
Subject: Re: Leica SP5 question
Hi Andy,
To be able to do XYZT acquisition with multiple fields your
scope needs to be fitted with a motorised stage. If you have that then
you will be fine.
To set up mark your first position, give it a z range and press the
exchange position button. If you don't hit the exchange position button
all your z stacks will be based on the last settings you give it.
I have done XYZT experiments in the past that have been 10-15 Z planes
over 24 different stage positions.
Cheers
Cam
Cameron J. Nowell
Microscopy Manager
Centre for Advanced Microscopy
Ludwig Institute for Cancer Research
PO Box 2008
Royal Melbourne Hospital
Victoria, 3050
AUSTRALIA
Office: +61 3 9341 3155
Mobile: +61422882700
Fax: +61 3 9341 3104
Facility Website
-----Original Message-----
From: Confocal Microscopy List [mailto:
[hidden email]]
On Behalf Of Aryeh Weiss
Sent: Tuesday, 10 February 2009 4:47 PM
To:
[hidden email]
Subject: Re: Leica SP5 question
First -- thanks for the quick responses. We will ask our rep to get us
the
software upgrade (the system is only a few months old -- I assume that
getting
the software will not be a problem).
Concerning mark and find with live data mode,
I think that it is not available on our machine. This is a problem
because we
have not been able to find a way to do a multipoint timelapse
acquisition.
Maybe the experienced SP5 users can correct me on that one. Is there a
way to
set up an XYZT acquisition with multiple fields?
--aryeh
Adrian Smith wrote:
> Also, does the Mark & Find function work in Live Data in version 2?
>
> When I had a quick look at in 1.8.2 it didn't seem like "mark and
find"
> was available in Live Data mode? (or at least I couldn't see it).
>
> Regards,
>
> Adrian Smith
> Centenary Institute, Sydney, Australia
>
>
>
> On 10/02/2009, at 2:32 PM, Dr. Graham Wright wrote:
>
>> Hello Zoltan & listers,
>>
>> You comments suggest that you are a regular user of the Leica Live
>> Data Mode on the SP5... if so, I was wondering how you handle the
data
>> post-acquisition - to get back to individual time series from each of
>> the different positions. We tend to use ImageJ and macros we have
>> written to sort and concatenate the files, but I am interested in how
>> others approach this.
>>
>> Best wishes,
>> Graham
>>
>> ----- Original Message -----
>> *From:* Zoltan Cseresnyes <mailto:
[hidden email]>
>> *To:*
[hidden email]
>> <mailto:
[hidden email]>
>> *Sent:* Tuesday, February 10, 2009 6:36 AM
>> *Subject:* Re: Leica SP5 question
>>
>> Aryeh,
>>
>> If you upgrade to LAS AF version 2.0, you'll be able to open a
>> second image window and observe previously acquired data. We very
>> much like the new version anyway, e.g. we find the Mark and Find
>> and Live Data Mode more stable than even with the 1.8.2, which
was
>> already pretty good.
>> I hope this helps,
>>
>> Zoltan
>>
>>
>>
>> On Mon, Feb 9, 2009 at 10:31 PM, Aryeh Weiss <
[hidden email]
>> <mailto:
[hidden email]>> wrote:
>>
>> When acquiring confocal data, we find it useful to have a
>> previous experiment (usually an image acquired earlier during
>> that session) open, while making the live acquisition. This
>> allows an immediate visual comparison of the current result
>> with a previous result. On the Leica SP5, we do not seem to
be
>> able to do this, because apparently only one data window can
>> be opened, and if a new scan is done, it replaces the old
data
>> in that window.
>>
>> I suppose I can use ImageJ to open the stored data, but i was
>> wondering if we are missing something obvious.
>>
>> --aryeh
>> --
>> Aryeh Weiss
>> School of Engineering
>> Bar Ilan University
>> Ramat Gan 52900 Israel
>>
>> Ph: 972-3-5317638
>> FAX: 972-3-7384050
>>
>>
>>
>>
>> --
>>
>> Zoltan Cseresnyes
>> Facility manager, Imaging Suite
>> University of Cambridge, UK
>
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