Axiovert 200M Stage Calibration and MM

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Axiovert 200M Stage Calibration and MM

O'Hare, Peter F

Hello

 

I wonder if anyone can help and apologies if this is not correct route

 

I am trying to use MM with an Axiovert 200M  and via MCU28 for stage control

 

All is actually going okay, regarding interface control, channel, motorised nosepiece etc;   the Pixels are calibrated and using the live image and graticule,   measurements of image, spot on.     I can also move the stage with the Device Control>Stage,  and it evens says the correct field width in um in the “Field” window

 

BUT, when I actually come to move the stage a set distance (or a field),  (or try to use HCS to take eg 96 wells) this is not working for stage movement.  So the actual image dimensions are okay, but it seems to send wrong pulses to MCU28 to move the desired amount

 

This precise problem (I think identical, as if hard wired via MM and MCU28) was in a post and reply from Nico,  but I cant see if it was solved

https://sourceforge.net/p/micro-manager/mailman/message/32075264/

 

This was the link in  MM forum, and text is copied here below.

Can anyone help?   Grateful for any advice or pointers

 

Thank you and apologies again if not correct route for assistance

 

Peter

 

Professor P O’Hare

Section of Virology

Faculty of Medicine

Imperial College

London W2 1PG

Tel. 0207 5949517

 

 

Hi Martin,
 
>  I am having a stage calibration issue with my Zeiss XY stage, driven by a MCU-28. When I tell MM to move the stage by 100µm the display tells me it's moving the right distance, but the stage actually moves by about 130µm, so there seems to be a mismatch (I measured the movement with a calibrated grid). If I use the Zeiss MTB program to move the stage it moves exactly 100µm. I started to noticed this when I tried to do tiling and there were large gaps between the images.
>  Any ideas on how to fix this?
 
The code to the MCU28 ( https://valelab.ucsf.edu/svn/micromanager2/trunk/DeviceAdapters/ZeissCAN/mcu28.cpp ) contains an interesting line:
 
stepSizeUm_(0.2) // assuming fixed step size at 0.2um      
 
A quick calculation shows that your system has a step size of 0.26 um.  I do not know if it is possible to automatically deduce the stepsize from the system, but we could add a property that lets you set the step size.
 
Best,
 
Nico

 

 



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Re: Axiovert 200M Stage Calibration and MM

Stuurman, Nico
Hi Peter,

I am trying to use MM with an Axiovert 200M  and via MCU28 for stage control

 

All is actually going okay, regarding interface control, channel, motorised nosepiece etc;   the Pixels are calibrated and using the live image and graticule,   measurements of image, spot on.     I can also move the stage with the Device Control>Stage,  and it evens says the correct field width in um in the “Field” window

 

BUT, when I actually come to move the stage a set distance (or a field),  (or try to use HCS to take eg 96 wells) this is not working for stage movement.  So the actual image dimensions are okay, but it seems to send wrong pulses to MCU28 to move the desired amount

 

This precise problem (I think identical, as if hard wired via MM and MCU28) was in a post and reply from Nico,  but I cant see if it was solved

https://sourceforge.net/p/micro-manager/mailman/message/32075264/


I checked the source code and there now indeed is a pre-initialization property for this.  You will need to figure out what the real step size is (probably best to tell the stage to move a certain distance while looking at a graticule to measure how far it actually moved, this will not tell you the real step size, but give you the ratio with respect to what it is set at now).  Then go into the hardware configuration wizard, select the MCU28, and look for a (pre-initialization) property called ""StepSize(um)".  It is likely set to 0.2 microns.  Multiply this by the factor you found, save the config file, and check whether things are now moving the correct distance.  You can also edit our config file in a text editor, and change this number manually (make a copy of the file before, in case you end up making mistakes).

Best,


Nico





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