Abstract
In this study, a rapid and simple method which combines drop coating deposition and Raman spectroscopy (DCDR) was developed to characterize the dry embryo culture media (ECM) droplet. We demonstrated that Raman spectra obtained from the droplet edge presented useful and characteristic signatures for protein and amino acids assessment. Using a different analytical method, scanning electron microscopy coupled with energy dispersive X-ray analysis, we further confirmed that Na, K, and Cl were mainly detected in the central area of the dry ECM droplet while sulphur, an indicative of the presence of macromolecules such as proteins, was mainly found at the periphery of the droplet. In addition, to reduce sample preparation time, different temperatures for drying the droplets were tested. The results showed that drying temperature at 50°C can effectively reduce the sample preparation time to 6 min (as compared to 50 min for drying at room temperature, ∼25°C) without inducing thermal damage to the proteins. This work demonstrated that DCDR has potential for rapid and reliable metabolomic profiling of ECM in clinical applications.
Highlights
Assessment of oocyte embryo quality helps identify the embryo with highest likelihood of implantation and the greatest pregnancy potential
Recent studies have suggested that metabolomic profiling of embryo culture media (ECM) can identify human embryos
To the best of our knowledge, we demonstrate for the first time, the application of drop coating deposition and Raman spectroscopy (DCDR) for the rapid and sensitive detection of ECM
Summary
Assessment of oocyte embryo quality helps identify the embryo with highest likelihood of implantation and the greatest pregnancy potential. The embryos selected for transfer are chosen according to morphological criteria and rate of development in culture on microscopic assessment, which is considered to be simple and practical over other methods for scoring the oocytes and embryos. Morphological evaluation is noninvasive, the problem is that relying solely on morphology assessment may not accurately predict the ability of an embryo to implant[1]. Morphology assessment always requires skilled personnel and is difficult to standardize. Methods that can provide a reliable and objective assessment of a given embryo are highly desired. Recent studies have suggested that metabolomic profiling of embryo culture media (ECM) can identify human embryos
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