Light intensity and wavelength during embryo manipulation are important factors for maintaining viability of preimplantation embryos in vitro  Seo Jin Oh, B.Sc., Seung Pyo Gong, B.Sc., Seung Tae Lee, Ph.D., Eun Ju Lee, Ph.D., Jeong Mook Lim, D.V.M., Ph.D.  Fertility and Sterility  Volume 88, Issue 4, Pages 1150-1157 (October 2007) DOI: 10.1016/j.fertnstert.2007.01.036 Copyright © 2007 American Society for Reproductive Medicine Terms and Conditions

Figure 1 Alignment of partial DNA sequence of Hsp70 of mouse (Genebank accession no. NM_008300) and hamster (Mesocricetus auratus, Genebank accession no. EF205024). Multiple sequence alignments were performed using CLUSTAL W (http://www.ebi.ac.uk/clustalw). Oh. Optimization of embryo culture system. Fertil Steril 2007. Fertility and Sterility 2007 88, 1150-1157DOI: (10.1016/j.fertnstert.2007.01.036) Copyright © 2007 American Society for Reproductive Medicine Terms and Conditions

Figure 2 Comparison of Hsp70 expression in hamster embryos manipulated under the light of different wavelengths. The Hsp70 mRNA levels were quantified by real-time PCR using an Applied Biosystems 7300 real-time PCR system and morulae retrieved 90 hours after mating. In total, five replicates were conducted, and the Hsp70 mRNA level in embryos cultured under visible (control, no filtering), blue, or red ray were provided. The mRNA level was normalized to that of β-actin mRNA and relative mRNA level was presented as 2−ΔΔCt, which enabled comparison with the level from the control group. Compared with no filtering, increased and decreased expression of Hsp70 was detected in embryos cultured under blue and red ray, respectively. Oh. Optimization of embryo culture system. Fertil Steril 2007. Fertility and Sterility 2007 88, 1150-1157DOI: (10.1016/j.fertnstert.2007.01.036) Copyright © 2007 American Society for Reproductive Medicine Terms and Conditions

Figure 3 Intracytoplasmic concentration of reactive oxygen species (ROS) in hamster embryos manipulated under the light of different wavelengths. ROS concentration was measured by dichlorohydrofluorescein diacetate (DCHFDA) fluorescent assay in morulae retrieved 90 hours after mating. Morulae were derived from culture under the lighting of visible (light from electric bulb of stereo microscope in dark room), blue, or red rays. A significant (P<.0001) increase in ROS concentration was detected with the blue ray compared with the visible control. a-cSignificant difference (P<.0279). Oh. Optimization of embryo culture system. Fertil Steril 2007. Fertility and Sterility 2007 88, 1150-1157DOI: (10.1016/j.fertnstert.2007.01.036) Copyright © 2007 American Society for Reproductive Medicine Terms and Conditions