Scaling
Scaling is a measure of how many microns a pixel represents. Scaling may be different for each axis and is automatically defined for the X and Y axes of each lens when you calibrate the system (Calibrating your system). If you're working with an image stack, the Z scaling (also known as the Z step) is equal to the distance between images.
Scaling is embedded for:
- Images acquired on a properly calibrated MBF system and saved in an MBF format (e.g., MBF .TIF or MBF .JP2).
- Multiple third-party image formats (e.g., ICS/IDS, LIF, and LSM).
Scaling is generally NOT embedded for TIFF files. Make sure that the proper scaling is applied to obtain valid measurements.
How do I apply the proper scaling?
You need to know the scaling used at the time of acquisition.
- Load the image. The Image Scaling dialog box opens.
- Check the Override X and Y scaling checkbox.
- Under Source of Scaling, select User Specified.
- Enter the correct values for X and Y.
- Neurolucida prompts you to define a new lens. Click Yes.
- Neurolucida displays an alert. Click Yes.
- The Create Lens from Image File dialog box appears.
- Make adjustments as needed.
Tracing from images without the correct scaling may result in invalid data.
Creating a lens or Creating a new lens based on a calibration grid slide