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Clicking on the little arrow on the right of a layer in the Layers palette opens a drop-down list where you can pick a colour from red, green and blue to apply to the layer. Once colours have been applied a new merge layer can be made by selecting the layers you want to merge and dragging them to the New button at the top of the palette or selecting Merge to RGB from the Layers menu.

A bright field image can be merged with fluorescence by placing it above the fluorescence layer in the palette, changing its opacity and then selecting Merge to Composite from the Layers menu. Colours other than RGB can be applied using the colour swatch in the document window tool bar.

Click on Image in the main menu and select 'Calibration...' to open the Spatial Calibration dialogue box. Click the unobtrusive little arrow icon on the right of the box (ringed in red in the adjacent image) to expand a list of saved calibrations. These have been measured by Light Microscopy Facility staff members so they may only include the most popular lens and camera binning combinations. If you want to use a different magnification factor (e.g. 4x binning, 2.5x optovar lens, etc.) then please email a request to the light microscopy facility. Select a saved calibration and click Calibrate. All images in the document will now be calibrated.

IMPORTANT: All the images in the document will have the same calibration so if you have changed magnification or binning while capturing to the same document at least some of your calibrations will be wrong. The best thing to do is to make sure all images in a document are captured at the same magnification and to open a new document if you want to change magnification.