Raman spectroscopy for live early mammalian embryos: optimization of experimental conditions and selection of the estimation criteria
E. Perevedentseva 1,2, A. Karmenyan 1, A. Krivokharchenko 3, M. Sarmiento 1, E. Baru 1, S. Manuel 1, V. Nadtochenko 3,4, C.-L. Cheng 1
1 National Dong Hwa University, Hualien, Taiwan
2 P.N. Lebedev Physical Institute RAS, Moscow, Russia
3 N. Semenov Federal Research Center for Chemical PhysicsRAS, Moscow, Russia
4 Department of Chemistry, Lomonosov Moscow State University, Moscow, Russia
Abstract
Laser spectroscopy is a promising tool in investigating and evaluating early mammalian embryos with direct measurements from the analyzed embryo but low invasiveness. Laser excitation wavelength is a key parameter in terms of safety and is important criteria for embryo estimation. In this work we present our search of ways to estimate the state of early developing mammalian embryo using Raman spectra obtained from different morphological areas of the embryo at different laser excitations.
Previously we have shown that using 488 nm wavelength laser excitation stimulates significant disturbance on the molecular level in the just extracted 2-cell embryo and completely interrupts the embryo development. Using 532 nm wavelength laser excitation allows selecting conditions in a narrow range of laser power and exposure time for safe measurements; following the normal development embryo in-vitro up to blastocyst forming. The advantage of using 532 nm excitation is the resonance Raman scattering of Cytochrome C. We have found the Cytochrome C Raman signature is strongly dependent to the embryo state; thus, it can be used for the embryo state analysis. Excitation using 785 nm wavelength laser falls into the tissue transparency window and expectedly to be the safest for the embryo. We compare the facilities of the embryo analysis using 532 and 785 nm excitations, and the optimal conditions to obtain appropriate information, perspectives of statistical methods of analysis. The possibility of using Raman spectroscopy for studying and evaluation of early live mammalian embryo are demonstrated.
Speaker
Elena Perevedentseva
Physics Department, National Dong Hwa University
Taiwan
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