**5.2 Microscopy—clinical samples**

Typical skin smear samples were collected before the supplementation and 4 weeks after [5]. The duplicates of the specimens were fixed, stained and analysed: the first one for lycopene with fluorescein isothiocyanate conjugated monoclonal mouse antibodies, at 200× magnification, and the second for lipid droplets of the sebum with Oil Red O, at 1000× magnification, as described previously [6].

### **5.3 Melting and defrosting time**

Approximately 20 mg of butter, or pork fat, or beef fat, or cocoa butter with or without certain concentrations of carotenoids, were placed on the surface of the laboratory slides and incubated at 37°C in a laboratory incubator (TLK39) until they melted.

To determine defrosting time, frozen oil aliquots of 200 mL were incubated at an ambient temperature of 20±2°C until they were completely melted. The period of time required for the tested sample to be melted or defrosted was measured with a laboratory timer (QUANTUM).
