Aberrant behavior of mouse embryo development after blastomere biopsy as observed through time-lapse cinematography Tomohisa Ugajin, M.D., Yukihiro Terada,

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Aberrant behavior of mouse embryo development after blastomere biopsy as observed through time-lapse cinematography Tomohisa Ugajin, M.D., Yukihiro Terada, M.D., Hisataka Hasegawa, M.S., Clarissa L. Velayo, M.D., Hiroshi Nabeshima, M.D., Nobuo Yaegashi, M.D. Fertility and Sterility Volume 93, Issue 8, Pages 2723-2728 (May 2010) DOI: 10.1016/j.fertnstert.2009.06.056 Copyright © 2010 American Society for Reproductive Medicine Terms and Conditions

Figure 1 Procedure of blastomere removal. We used a piezo-driven pipette for blastomere removal. (A) The zona pellucida was penetrated with a piezo pulse. (B) After expulsion of the segment of zona pellucida within the pipette, the pipette was reinserted, and one blastomere was aspirated. (C) A three-cell embryo after blastomere removal. Bar = 40 μm. Fertility and Sterility 2010 93, 2723-2728DOI: (10.1016/j.fertnstert.2009.06.056) Copyright © 2010 American Society for Reproductive Medicine Terms and Conditions

Figure 2 The timing of compaction after blastomere removal. The embryo with one blastomere removed initiated compaction at the six-cell stage, after three rounds of cell division. Bar = 20 μm. Fertility and Sterility 2010 93, 2723-2728DOI: (10.1016/j.fertnstert.2009.06.056) Copyright © 2010 American Society for Reproductive Medicine Terms and Conditions

Figure 3 Stages of embryonal development after blastomere removal. We calculated the time between blastocele formation and the end of hatching and the time between the start and end of hatching. (A) An embryo after blastocele (circle) formation. Bar = 20 μm. (B) An embryo in the process of hatching. Bar = 20 μm. (C) An embryo after hatching. Bar = 20 μm. Fertility and Sterility 2010 93, 2723-2728DOI: (10.1016/j.fertnstert.2009.06.056) Copyright © 2010 American Society for Reproductive Medicine Terms and Conditions

Figure 4 Contraction and expansion of a mouse blastocyst. Mouse embryos underwent contraction and expansion after blastocyst formation and before hatching. (A) An embryo in the process of expansion. (B) An embryo in the process of contraction. Bar = 20 μm. Fertility and Sterility 2010 93, 2723-2728DOI: (10.1016/j.fertnstert.2009.06.056) Copyright © 2010 American Society for Reproductive Medicine Terms and Conditions

Figure 5 Measurement of maximum diameter of a mouse embryo. The maximum diameter of expanded blastocysts just before hatching was calculated as the average of the major and minor axis lengths. Bar = 30 μm. Fertility and Sterility 2010 93, 2723-2728DOI: (10.1016/j.fertnstert.2009.06.056) Copyright © 2010 American Society for Reproductive Medicine Terms and Conditions