Effect of varying equilibration time in a two-step vitrification method on the post-warming DNA integrity of mouse blastocysts Amr Kader, M.D., Audrey.

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Presentation transcript:

Effect of varying equilibration time in a two-step vitrification method on the post-warming DNA integrity of mouse blastocysts Amr Kader, M.D., Audrey Choi, B.A., Rakesh K. Sharma, Ph.D., Tommaso Falcone, M.D., Ashok Agarwal, Ph.D., H.C.L.D. Fertility and Sterility Volume 93, Issue 8, Pages 2640-2645 (May 2010) DOI: 10.1016/j.fertnstert.2009.07.991 Copyright © 2010 American Society for Reproductive Medicine Terms and Conditions

Figure 1 Imaging with confocal Leica software. (A) Different sections of the blastocyst in the blue fluorescence channel. (B) Different sections of the blastocyst in the green fluorescence channel. (C) Different sections of the blastocyst in both the blue and the green fluorescence channels merged. (D) Overlay image of the blastocysts in the blue fluorescence channel. (E) Overlay image of the blastocysts in the green fluorescence channel. (F) Overlay image of the blastocysts in both the blue and the green fluorescence channels merged. Scale bar = 10 μm. Fertility and Sterility 2010 93, 2640-2645DOI: (10.1016/j.fertnstert.2009.07.991) Copyright © 2010 American Society for Reproductive Medicine Terms and Conditions

Figure 2 Calculating the DNA integrity index by counting the blastomeres using “Volocity 64” software. TUNEL− blastomeres are blue; TUNEL+ blastomeres are green. (A) Start of counting at the superficial layer of the blastocyst. (B) Continuous counting at a deeper cross-section. (C) Continuous counting at an intermediate cross-section. (D) Counting at a bottom layer of the blastocyst. (E) Counting of the TUNEL+ cells, which are marked with a red arrow for better visualization. (F) Counting of TUNEL+ TUNEL− blastomeres all gathered in the “Volocity 64” software with information on each blastomere location. Scale bar = 10 μm. Fertility and Sterility 2010 93, 2640-2645DOI: (10.1016/j.fertnstert.2009.07.991) Copyright © 2010 American Society for Reproductive Medicine Terms and Conditions

Figure 3 Median DNA integrity indices in the fresh and vitrified nonexpanded and expanded blastocysts after 4, 8, or 15 minutes' equilibration time. The red line connects the median DNA integrity indices of the nonexpanded blastocysts. The blue line connects the median DNA integrity indices of the expanded blastocysts. Fertility and Sterility 2010 93, 2640-2645DOI: (10.1016/j.fertnstert.2009.07.991) Copyright © 2010 American Society for Reproductive Medicine Terms and Conditions