The script has been preconfigured for specific dye combinations. For the un-mixing to work, the correct combination must be selected. Immediately below the Input Channel settings, you'll find the STAINING settings.

# -------------------------------------------------------------------------

# 0 == HEMA_DAB

# 1 == HEMA_EOSIN

# 2 == HEMA_EOSIN2

# 3 == FEULGEN_LGRENN

# 4 == HEMA_EOSIN_DAB

# 5 == GIEMSA

# 6 == Fast Red, Fast Blue and DAB

# 7 == Methyl green and DAB

# 8 == Haematoxylin, Eosin and DAB (H&E DAB)

# 9 == Haematoxylin and AEC (H AEC)

# 10 == Azan-Mallory

# 11 == Alcian blue & Haematoxylin

# 12 == MASSON THRICROME

# 13 == Haematoxylin and Periodic Acid of Schiff (PAS)

# -------------------------------------------------------------------------

STAINING = 0

Simply replace the number after the "STAINING = " to match your image. For example, if using Hematoxylin and Eosin (H&E), the line should then read:

STAINING = 1