Re: LSM510 multiple locations macro-detailed image
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Julio Vazquez |
Re: LSM510 multiple locations macro-detailed image
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Hi Yi,
I wrote the LSM 510 manual a long time ago, and will need to check if the instructions for the multi-time macro are correct. I will send you a screen capture of the Multi-Time window with all the settings for imaging at multiple locations. What you need to do is: (a) mark the different locations with the stage control panel, (b) create the configuration you need for imaging your sample, and test it to see it's giving you good results... note that you should be in Multi-track mode. Note also that all the channels (tracks) you need to use are checked (active) in the configuration control panel before you save it, (c) load the multi-track configuration into the Multi-Time control panel, and (d) load the locations with the edit location tool in the Multi-Time panel. I generally select individual locations in the location list in the Multi-time panel and verify that the right configuration is associated with it.. I think the problem is that if you load the list of locations first, they will be linked to whatever configuration was remembered from a previous experiment. If you then load your new configuration, it will be applied only to the current location displayed at the top of the Multi-Time window, in which case you need to select each location and reload the right configuration for all of them The pinhole error message just indicates that the pinhole was never aligned for the specific track you were using, so either you were using a bad track, or you need to align the pinhole for the track you are using. If you got images at all locations as you were planning to, I suspect the configuration you used was not set properly, or was not applied all locations. In that case, follow the steps above and make sure the configuration you saved is working. If the macro itself did not run properly (confocal did not go to the desired locations, or do the right number of cycles, make sure your Multi-Time control panel is set-up as in the screenshot I will send you. good luck, Julio. -- Julio Vazquez Scientific Imaging Fred Hutchinson Cancer Research Center Seattle, WA 98109-1024 Tel: Office: 206-667-1215/ Lab: 206-667-4205 FAX: 206-667-6845 On Aug 25, 2008, at 9:34 PM, Yi Zheng wrote:
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Search the CONFOCAL archive at
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While I don’t qualify as a multitime expert (more of a dabbler),
I would only add the Zbignew Iwinsky (the programmer at Zeiss) frequently
updates the multitime macro. I believe the updates are in the user area
of the Zeiss FTP site (see instructions at: ftp://lsm.zeiss.com/The%20Zeiss%20LSM%20FTP.pdf) 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: [hidden email] voice: 520-626-2824 fax: 520-626-2097 http://swehsc.pharmacy.arizona.edu/exppath/ Home of: "Microscopy and Imaging Resources on the WWW" From: Confocal Microscopy
List [mailto:[hidden email]] On Behalf Of Julio Vazquez Search the CONFOCAL archive at
http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal Hi Yi, I wrote the LSM 510 manual a long time ago, and will need to
check if the instructions for the multi-time macro are correct. I will send you
a screen capture of the Multi-Time window with all the settings for imaging at
multiple locations. What you need to do is: (a) mark the different
locations with the stage control panel, (b) create the configuration you need
for imaging your sample, and test it to see it's giving you good results...
note that you should be in Multi-track mode. Note also that all the channels
(tracks) you need to use are checked (active) in the configuration control
panel before you save it, (c) load the multi-track configuration into the
Multi-Time control panel, and (d) load the locations with the edit location
tool in the Multi-Time panel. I generally select individual locations in the
location list in the Multi-time panel and verify that the right configuration
is associated with it.. I think the problem is that if you load the list of
locations first, they will be linked to whatever configuration was remembered
from a previous experiment. If you then load your new configuration, it will be
applied only to the current location displayed at the top of the Multi-Time
window, in which case you need to select each location and reload the right
configuration for all of them The pinhole error message just indicates that the pinhole
was never aligned for the specific track you were using, so either you were
using a bad track, or you need to align the pinhole for the track you are
using. If you got images at all locations as you were planning to, I suspect
the configuration you used was not set properly, or was not applied all
locations. In that case, follow the steps above and make sure the configuration
you saved is working. If the macro itself did not run properly (confocal did not
go to the desired locations, or do the right number of cycles, make sure your
Multi-Time control panel is set-up as in the screenshot I will send you. good luck, Julio. -- Julio Vazquez Scientific Imaging Fred Hutchinson Cancer Research Center Seattle, WA 98109-1024 Tel: Office: 206-667-1215/ Lab: 206-667-4205 FAX: 206-667-6845 = On Aug 25, 2008, at 9:34 PM, Yi Zheng wrote:
Search the CONFOCAL archive at Here are detailed images: [/IMG] [/IMG] |
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