Re: Leica SP5 question

Posted by Zoltan on
URL: http://confocal-microscopy-list.275.s1.nabble.com/Zeiss-25-X-Plan-Neofluor-Phase-3-objective-for-sale-tp2286535p2305957.html

Aryeh,

We either run Mark and Find, or Live Data Mode.  In the latter case, each Job corresponds to a Z stack at a certain position on the slide, all parameters set up to best conditions.  We then add all Jobs to a macro with appropriate time gaps in between consecutive Jobs (Wait/Pause option), and then connect the two ends of the macro and set up the appropriate number of loops.  This we do usually for overnight live cell/animal scanning.  

  With Mark and Find, we choose XYZT mode, visit every location that we want to scan on the slide one-by-one, set up all scan parameters at a given position and THEN (at the end!) we save the position; this way the system will remember all the settings. Then we move to the next position and repeat.  As others pointed out already, at the beginning you have to have your motorised stage initalised for both the Live Data Mode and the Mark and Find.  In Mark and Find you also have to unclick the "Use same Z stack" option box and must click on the Set Stack (it's called something like this) software button before you move on to the next position.         

  I hope this helps,

Zoltan

- Show quoted text -

On Tue, Feb 10, 2009 at 5:46 AM, Aryeh Weiss <[hidden email]> wrote:

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: [phone]




   --
   Zoltan Cseresnyes
   Facility manager, Imaging Suite
   University of Cambridge, UK

--

Zoltan Cseresnyes
Facility manager, Imaging Suite

University of Cambridge, UK