Regarding the resetting of a zero position: On a different instrument where
we did automated analysis of 96 well plates, we created a fiduciary mark on
a blank plate and set that as zero every time we ran the analysis. All the
measurement positions were created from that initial point. To make the
mark we scratched a cross-hair on the plate's bottom surface using a sharp
razor blade.
Doug
^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^^
Douglas W. Cromey, M.S. - Assistant Scientific Investigator
Dept. of Cell Biology & Anatomy, University of Arizona
1501 N. Campbell Ave, Tucson, AZ 85724-5044 USA
office: AHSC 4212 email: [log in to unmask]
voice: 520-626-2824 fax: 520-626-2097
http://swehsc.pharmacy.arizona.edu/exppath/
Home of: "Microscopy and Imaging Resources on the WWW"
-----Original Message-----
From: Confocal Microscopy List [mailto:[log in to unmask]] On
Behalf Of Danielle Crippen
Sent: Friday, June 19, 2009 9:08 AM
To: [log in to unmask]
Subject: Re: Locations in MultiTime macro
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
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