For more advanced image acquisition configurations with multiple positions set up your channels as described in SIMPLE CONFOCAL IMAGE ACQUISITION. Then open the Multi-Area Time-Lapse controller, which can be found under Device -> Multiarea Time-Lapse.
An XY-stage calibration warning will open. Press OK to calibrate the stage, which is important to be able to register points for multipoint acquisitions.
If you would like to use the Z-Drift Controller (ZDC) to keep the focus stable throughout the experiment follow the instruction in the Z-DRIFT CORRECTION (ZDC) section. Be aware that you will need to restart the software to set this up.
In Multiarea Time-Lapse Controller:
1. 1^st button from left: Add current position (to save a position, then move on to next -> gives you list of data points in Registered Point List)
2. 2^nd button from left: Mosaic Outline (to select separate points and then fills in gaps between them to scan an area)
3. 3^rd button from left: Define Matrix (specify central point and define area around it)
Go to View -> Stage controller to control the stage using the software or use the manual XY-controller to navigate to different positions in your sample and create a point map using the Add Current Position, Mosaic Outline or Define Matrix method.
Double-click on a number in the Registered Point List to go to positions you marked previously.
In the Add Current Position mode it is possible that the added points have different Z-positions.
The Interval and Repeat field allow you to set-up time series.
It is also possible to combine the multi-point acquisition with a Z-stack.
To reload a previous point list navigate to Device -> Multi-Area Time-Lapse Viewer -> Open files in the main window.
To clear points go to the Registered Point List, highlight points you do not want to keep and press the trash button.
When you want to acquire a Z-stack or time-series you need to go to Edit -> Edit acquisition parameters and select Depth and/or time respectively. Then press OK. Be aware that this only activates the Z-stack for the currently selected position and needs to be activated for all other positions separately.
If you make changes to a marked position and want to update it go to Edit -> Re-register.
Before starting the image acquisition create a location on the PC hard disk to save your data to by navigating to Options -> File & Folder -> Folder name (you can also create a subfolder for each data point).
When you are happy with the experiment set-up press the Ready button (bottom left) in the Registered Point List window.
Press Start to start the image acquisition.
To obtain a stitched image from raw Mosaic Outline or Define Matrix data go to Stitch View -> select central plane if Z-stack -> Show -> Save as series.

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