Confocal laser scanning microscopy was conducted using an
inverted Leica SP5 microscope (Leica Microsystems, Wetzlar,
GmbH, Germany) equipped with krypton/argon and helium/
neon lasers. A water immersion objective was used for 63
magnification. The dyes were added on a pre-cooled object glass
one at a time when the former dye solvent had evaporated.
Next, a chunk of the butter was placed on the objective glass and
left to equilibrate for 30 min at 5 8C prior to analysis (Rønholt
et al., 2013c). Sequential scanning was used for all samples