Abstract:Objective To develop a method with Raman spectroscopy for selecting embryos with high developing potential. Methods A total of 100 samples of D3 spent cultural medium for embryos assigned to blastocyst culture were collected,and the Raman spectroscopy produced with optimal integration time. The original spectrums retrieved were processed by filtration,baseline correction and normalization methods to form characteristic spectral for the spent medium of cleavage embryo,suitable for analysis. A further 173 samples were collected to analyze the clinical relevance of Raman spectroscopy in selecting the potential embryos. Results Original Raman spectra with good resolution can be obtained with an integral time over 240 seconds. The characteristic spectral shift of embryo culture medium was 942,1 346,1 456 and 1 656 cm-1 after processing. The spectral shift between the positive curve and the negative model was 1 100-1 800 cm-1. There was no significant difference in the intensity of real Raman spectral with different shifts(P>0.05). A total of 80 positive curves,70 negative curves,23 neutral curves were obtained from the 173 D3 samples,and the blastocyst formation rates were 85.00%,22.86% and 65.22%,respectively,with statistically significant difference(P<0.001). The sensitivity,specificity and Kappa value for the Raman spectrum curve predicting the blastocyst formation were 83.8%,73.0% and 0.572,respectively. Conclusion A clear,real and comparable Raman spectrum model could be obtained by improving the spectral collection conditions and preprocessing the original Raman spectrum. The potential embryo screened by this method is moderately consistent with that of blastocyst.
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