Confocal Microscopy List

Re: Leica SP5 question

Posted by George Ring on
URL: http://confocal-microscopy-list.275.s1.nabble.com/Zeiss-25-X-Plan-Neofluor-Phase-3-objective-for-sale-tp2286535p2303334.html

Hi, Aryeh,

As Cam said below, to do an XYZT on multiple fields you need a motorized
stage. Also, when the LAS AF boots up, you have to select "Yes" when the
application asks you if you want to initialize the Motorized Stage. Even
though the stage initializes when the microscope is turned on, you have to
initialize it again in order to load the tile scan and the multipoint
capabilities.

Hope this helps,

George

George Ring, Ph.D.
Dept. of Cell and Developmental Biology
SUNY Upstate Medical University
750 E. Adams St.
Syracuse NY 13210
Tel. (315) 464-8595
FAX (315) 464-8535
email: [hidden email]

>>> On Tue, Feb 10, 2009 at 12:59 AM, in message
<[hidden email]>, Cameron
Nowell <[hidden email]> wrote:

> 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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