How to set up a simple acquisition with an XY stage in Volocity

XY stage support is now available in Volocity, which means that you can use Volocity for multi-point acquisition experiments on slides or multi-well plates. Here I will cover how you set up the stage add points and perform a simple acquisition.

Start Volocity, go to the "Stage" menu and select "XY Stage Setup"

Select the XY Stage Device. If you have a focus device remember to select this as well. If you fail to do this the focus position will not be saved when you add points.

Click "Save" when you are happy with the setup.

The next step is to calibrate your stage, you should do this each time you start Volocity or turn on the controller.

Go to the "Stage" menu and select "Calibrate Stage". When you do this you will see the warning prompt below.

Make sure that the stage area is clear before you proceed, remove any samples and ensure that the focus drive is moved to a safe position to avoid damage to the objective lenses.

Once you have calibrated the stage you can use the XY stage view.

Go to the "Video" menu and select "XY Stage".

The crosshairs represent the current stage position. If you move the stage the crosshairs will move to reflect the change in position. To quickly move the stage to a new position select the rectangular ROI tool and double click on the point in the XY Stage work area.

A small video preview is shown in the bottom left hand corner of the view. To move it to a different corner simply click and drag it to the new position.

Before adding points I recommend that you focus on you sample and select "Set Zero" from the focus drive drop down menu.

Now you are ready to add points. Move the stage to the point of interest - you can use either the video preview or watch your sample position using the microscope eyepieces.

Once you are happy with the position go to the "Stage" menu and select "Add point".

Alternatively you can add a point using the shortcut key:

On a PC:  Ctrl+Shift+A

On a Mac: Shift+command+A

Move the stage and continue to add points as required. Each point is shown as a red cross in the XY Stage view. The X,Y and Z co-ordinates of each point are displayed next to the point, you will probably have to zoom in to see them.

If you would like to inspect any of the points that you have set double click on them in the XY stage view. This will move the stage and the focus drive to the X, Y and Z position of the point.

To change the focal position of a point move to the new focus position and select "Update Point" from the "Stage" menu.

If you have a lot of points that you would like to review select "Review Points" from the "Stage" menu.  The stage and focus drive will move to each reviewed point, at each point change the focus position if required then click "Next", the current focus position will be stored.

Finally we need to setup the acquisition protocol. Go to the "Video" menu and select "Acquisition Setup".

Set up the acquisition protocol as required, as you can see I have decided to use two light paths.

Make sure that you select the XY stage from the popup menu. As I am not acquiring a Z series I have not selected a focus drive from the drop down menu. If you wish to acquire a z series please ensure that you select the required focus drive.

You will notice that you can use auto-focus for your points during acquisition. If you require this option select the light path that you would like to use for auto-focusing.

If you select this option you can also set timing options by clicking on the "timing" button.

You can choose to focus at each point and set the frequency of the auto-focus as shown in the dialog below.

Once you are happy with the Acquisition setup click the "OK" button. In the acquisition controls set up the time lapse options.

I have chosen to acquire a time point every five minutes.

Once you are ready click on the "Start" button to commence the acquisition.

A separate image sequence will be acquired for each point.

I hope this provides a taster into using the XY stage with Volocity. I have only scratched the surface here and there are many more options.  Please consult the Volocity User Guide for further reading.