Re: LSM510 multiple locations macro-detailed image
Posted by
Julio Vazquez on
URL: http://confocal-microscopy-list.275.s1.nabble.com/Re-LSM510-multiple-locations-macro-detailed-image-tp783186p784740.html
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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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Here are detailed images:
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