Multi-point Acquisition

Point by Point (Manual) Multi-Point
Well Plate (Rectangular) Multi-Point
Large Image (Covering) Multi-Point
Random Multi-Point
To Change a Single Z Coordinate
Special Options
Advanced Settings

This window serves for defining XY(Z) points to be scanned during the multipoint capture experiment. This feature is available when a motorized stage XY(Z) is present in the system. Check the Include Z box to display the Z column. The list of defined points can be saved (and loaded later) to an XML file by the Save (Load) button. Run the Acquire > Capture Multipoint > Capture Automatically command.

Point by Point (Manual) Multi-Point

Move the stage to the first position.
Press the Add New button. A new line containing current coordinates of the stage appears in the list.
Move to the next position and repeat the steps until you have all the intended points defined.

Well Plate (Rectangular) Multi-Point

You can insert a pattern designed to cover a wellplate. Of course it can be used to create any rectangular pattern.

Click the Custom button. The following window appears
Figure 279.
Select Manual if you know the distances between wells. Otherwise select the Interactive and continue.
Manual
This method will create a pattern which will scan Rows x Columns fields. The Distance X and Y parameters specify distances between two fields. The scanning will begin at the current position.
Interactive
The interactive method lets you specify the number of fields to be scanned and starts a wizard where you will specify the top-left and the bottom-right corner of the well plate. The distances between wells are calculated automatically.
Click Finish.

Large Image (Covering) Multi-Point

A Multi-point which will cover certain area can be created:

Click the Custom button and select the Large Image tab
Figure 280.
Set the large image settings:
Scan Area
Select whether to define the Scan Area by number of fields or the actual field size.
Camera
Select the camera to be used. This field is enabled only if you have a dual-camera setup.
Objective
Select the objective for which the multi-point will be calculated. The resulting number of fields / area size changes depending on the selected objective magnification.
Overlap
Overlap of neighboring images in %.
Click Finish.

Random Multi-Point

Specify the area where a random set of points will be generated.

Click the Custom button and select the Random tab.
Figure 281.
Select shape of the area to be filled with random points
Rectangle
The area will be defined by a rectangle. Define X and Y coordinates of left top corner of the area, width and height of the area.
Radius
The area will be defined by a circle. Define X and Y coordinates of center of the circle.
Specify Number of points in the multi-point set.
Click Finish.

To Change a Single Z Coordinate

Make sure the Move stage to selected point is pressed. This button ensures that the motorized stage moves to the coordinates of the currently selected point.
Click inside the line you would like to change.
Move the Z drive to the new position.
Click the <- button.

Note

The XY coordinates of one point cannot be adjusted (unless you delete it and add a new point).

Special Options

Offset All

The offset all button can shift the XY coordinates of all points in the same way:

Make sure the Move stage to selected point is pressed. This button ensures that the motorized stage moves to the coordinates of the currently selected point.
Select one point of the list.
Move the XY(Z) stage to a new position (define the offset).
Press the Offset All button.
The coordinates of all points change - the same shift which you made with the stage is applied to them.

Move Stage to Selected Point

This button ensures that the motorized stage moves to the coordinates of the currently selected point.

Point name

Displays the name of the point. You can change the name (default names are #1, #2 #3, etc.).

X, Y, Z

Displays X, Y and Z coordinates of the point. The arrow buttons -> <- assign the current XYZ position to the point. The Offset All X,Y,Z button shifts the XYZ coordinates of all points by the same offset which is defined as a difference between the current stage coordinates and coordinates of the selected point.

Include Z

If selected, also the Z coordinate will be taken into account during XY(Z) acquisition.

Note

This option is disabled if the Relative XY option is used.

Define Stimulation Laser Powers

If checked, Laser Power Settings dialog window appears when defining stimulation ROIs. This window defines the power of each laser line on each stimulation group.

Relative XY

Check the Relative XY item to consider all coordinates as relative with respect to the current stage position. Any of the points may be used as the reference point, just right-click it and select Set this point as a reference position.

Optimize

If the Optimize button is pressed, the system will re-order the defined points in order to minimize the XY stage trajectory.

Custom

Displays a tool for creating predefined multi-point patterns.

See Well Plate (Rectangular) Multi-Point, Large Image (Covering) Multi-Point, Random Multi-Point.

Advanced Settings

Redefine reference Z after Auto Focus

Check this option to redefine reference Z position after performing autofocus or PFS (if AF or PFS is turned on).

Leave PFS offset ON between points

Check this item to keep PFS on while moving between the points.

Autofocus

See Common ND Experiment Options.

Execute Command before Capture, Execute Command after Capture

See Executing macro commands during ND acquisition.

Use Focus Surface

Uses Focus Surface for capturing.

Split Multipoints

Select this box to perform the File > Import/Export > Split Multipoints command directly after the ND2 experiment.

Note

When you apply the Split Multipoints command on ND acquisition that contains only the XY dimension, the information about the ND dimensions in the File > Open dialog becomes unavailable.