Dear Gareth,

First of all, I'd recommend calling Zeiss and talking with Zbi, Maya or John about this.  You can get advice from them regardless of whether you're on service contract...call 1-800-633-6610.  I find them to be endless resources.  (BTW--I have no affiliation with Zeiss...just use their equipment a lot).

Secondly...I'm wondering...are you zero-ing the stage?  Do you have a stage with linear encoders on it for the best accuracy?  How are you ensuring you're re-placing your plate in the exact same location as when you originally marked positions.  If you're hoping to image the same ROI (with the same cells in it etc), even a 50-100um xy shift can ruin your coordinates.  We've had some good success here with 8 well chambered coverglass by zeroing the stage at a particular corner of a particular well.  Then when we re-load the chamber, we zero again with the same method...then we can successfully return to previously imaged coordinates for further imaging...even weeks later.  

I think in answer to your first question, you can of course always manually re-do locations by moving the stage to the well you want to image, selecting that location in the MT list, and clicking the "replace xyz" button.  Once you've corrected each location, you can Store/Apply again and overwrite the MT setup file.

I can't comment on your second question, b/c I don't utilize the blocks in MT (yet:)...so I'm not sure how or if this would work.

Best of luck!!

d


Danielle Crippen
Morphology and Imaging Core Manager
Buck Institute for Age Research
8001 Redwood Blvd
Novato, CA 94945
415-209-2046



-----Original Message-----
From: Confocal Microscopy List [mailto:[log in to unmask]] On Behalf Of Gareth Howell
Sent: Friday, June 19, 2009 4:31 AM
To: [log in to unmask]
Subject: Locations in MultiTime macro

Hi
I have two questions relating to the MT macro which I'm using for screening
96 well plates. Currently I manually set the positions relating to each plate and save them within the macro using the Store/Apply step. However when they are recovered some (~10%) are missing the wells and we loose data within the screens. So my questions are:
1) is it possible to edit the locations within the Store/Apply so that they are all accurate?
2) is it possible to save each co-ordinate as a seperate configuration, with the stage coordinates included, and run the screen as a series of blocks within the macro? I realise this would be initially very labour intensive but worth the effort in the long run.
Thanks
Gareth