FIGURE4| Celldamageandcryosurvivalofblastocysts.BovineembryoswereexposedtostandardSOF,controlFAFSOF,25 μ MC 18:0 or25 μ MC 18:1 duringday 1 until 5 of in vitro culture. The percentage of damaged cells re fl ects the total sum of apoptotic (TUNEL) and necrotic (EthD-1) cells, from fresh blastocysts (A) and those that survived the freezing and thawing procedure (B) as a percentage of the total cell number (Hoechst 33342), analyzed via confocal microscopy. A representative confocalimageofafrozenthawedblastocyst pergroupisshownforStandardSOF (C) ,FAF SOF (D) ,C 18:0 (D) and C 18:1 (E) ,withinblueDNA(Hoechst33342),in green apoptosis (TUNEL) and necrosis in red (EthD-1). Panel G shows the cryosurvival rates per group. Panel H shows the cryosurvival rates per for respectively non- expanded and expanded blastocysts per group. Bar =50 µm.Differentsuperscripts differsigni fi cantly.Asterix indicates asigni fi cantdifference betweenthe blastocysts classes within a group. For runs and total number see Scheme 1 (in the Materials and Methods