Re: Leica SP5 question - Motorised Stage

Posted by Zoltan on
URL: http://confocal-microscopy-list.275.s1.nabble.com/Zeiss-25-X-Plan-Neofluor-Phase-3-objective-for-sale-tp2286535p2305817.html

Adrian,

We didn't really have to alter the factory settings in order to get the tiling to work very well (some of my users scan entire locust brains or parts of butterfly wings, which require at least 3x3 tiling, up to 5x5).  We had no problem tiling and the LAS AF automatic stitching option worked very well (tiling overlap set to zero).  I'm home now but tomorrow I'll try to remember to read the parameters out for you.  However, the only critical step was to make sure that the scan field rotation is set such that the sample moves perfectly parallel to the horizontal (X) edge of the screen when we are turning the X controller joystick knob.  We select a bright spot in the sample, move it very close to the lower edge of the image screen and slowly turn the joystick knob (X) while watching the bright spot move; we then adjust the field rotation until the bright spot moves perfectly parallel to the lower edge of the screen.  In our case this angle turned out to be -0.7 degrees.  

  I hope this helps,

Zoltan

On Tue, Feb 10, 2009 at 11:01 PM, Adrian Smith <[hidden email]> wrote:

On 10/02/2009, at 4:59 PM, Cameron Nowell wrote:

 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.

While we are talking SP5 and motorised stage....

I've been wondering if anyone has a come up with a good procedure for setting up the calibration of the stage position, ie in the Stage Configuration window there are settings for Tile Scan overlap, origin offset sizes, merge settings that need to be set up in order for tile scans to work.

One of the students here got them quite close through a laborious trial-and-error process. Unfortunately, we need to reset them because the stage alignment got messed up at one point (when the stage cable got caught in the opening into the incubation enclosure).

Before I go through the process of doing it by trial and error or working out a more systematic way of doing it I was wondering if anyone else had any suggestions as to how best approach it?

Regards,

Adrian Smith
Centenary Institute, Sydney, Australia

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Zoltan Cseresnyes
Facility manager, Imaging Suite

University of Cambridge, UK