The Axioscan Advanced Spectral Unmixing module enables you to create new reference spectra. Here is an overview of steps for the creation of reference spectra:

1. Prepare your reference samples for scanning. For information on how to best prepare the samples, see Sample Preparation for Creating New Reference Spectra.
2. Acquire reference scans and prepare the images with the usual functionalities of ZEN slidescan. For specific remarks and notes, see Information for Acquisition and Preparation of Reference Scans.
3. To create your reference spectra, start the Calibrated Unmixing wizard, see Starting the Calibrated Unmixing Wizard.
4. In the first step, select the autofluorescence spectra, see Selecting Autofluorescence Spectra.
5. In the second step, select the dye spectra, see Selecting Dye Spectra.
6. In the third step, check the calculated dye spectra, see Checking the Calculated Dye Spectra.
7. Once you have created your reference spectra, you can use them for unmixing in your scan profile, just as other reference spectra, see Applying Unmixing in the Scan Profile.