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. 2018 Feb 13;8:2902. doi: 10.1038/s41598-018-21335-8

Figure 4.

Figure 4

Quality of in vitro cultured mouse embryos produced by intracytoplasmic sperm injection (ICSI). For this experiment, swim-up separated vs thermotaxis selected spermatozoa were compared. (a) Morphological evaluation 24 h (% reaching 2-cell stage) and 5 days (% reaching morula + blastocyst stage) after ICSI (mean ± SEM, n = 8 experiments, 160 total ICSIs per sperm type). Different letters indicate significant differences (P < 0.01 two–way ANOVA). (b) Morphological evaluation 5 days after ICSI of morula and blastocyst embryos (mean ± SEM). Different letters indicate significant differences (P < 0.01 two–way ANOVA). (c) Representative image of embryos 5 days after ICSI cultured in vitro. Nf: not fertilized; F: fragmented/dead embryo; M: morula; E: early blastocyst; Ex: expanded blastocyst; H: hatching blastocyst. (d) Representative image of an in vitro cultured embryo processed for terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL). Red fluorescence marks apoptotic/necrotic nuclei. Hoechst staining was used to counterstain nuclei (blue). Bar = 50 µm. (e) Representative images of 2-cell embryos stained with TO-PRO–3 Iodide for abnormal chromosome segregation assessment. Arrow: extranuclear DNA fragment; EC: empty cell; Pb: polar body; N: nuclei. Bar = 25 µm.