Imaging lipids in living mammalian oocytes and early embryos by coherent Raman scattering microscopy

Paola Borri*, Josephine Bradley, Iestyn Pope, Wolfgang Langbein, Karl Swann

*Corresponding author for this work

Research output: Chapter in Book/Report/Conference proceedingConference contributionpeer-review

Abstract

Many promising techniques proposed to monitor gamete developmental potential and quality are invasive and not realistically useful in clinical practise. Hence, there is increasing interest in the development of non-invasive imaging methods that can be applied to mammalian eggs and early embryos. Recent studies have shown that mammalian oocyte and embryo viability are closely associated with their metabolic profile, relying entirely on mitochondria as a source of ATP. Fatty acids, stored in intracellular lipid droplets, are an important source of ATP. We have recently demonstrated the use of Coherent Anti-stokes Raman Scattering (CARS) microscopy to allow chemically-specific, label-free imaging of lipid droplets, with high three-dimensional spatial resolution. Here, we summarize our main findings when using CARS to examine the number, size, and 3D spatial distribution of lipid droplets in mouse eggs and early embryos. Quantitative analysis showed statistically significant differences during oocyte maturation and early embryo development. Notably, CARS imaging did not compromise maturation or development. In mouse oocytes that had been subjected to alterations in mitochondrial metabolism we found that the spatial distribution pattern of lipid droplets was also altered. In addition, differences in the chemical composition of lipid droplets in living oocytes matured in media supplemented with different saturated and unsaturated fatty acids were detected using CARS hyperspectral imaging. We also imaged bovine oocytes, and found that lipid droplets appear to be larger and with less spatial aggregation than in mouse oocytes, possibly reflecting the fact that different species metabolise lipids differently. These data suggest that CARS microscopy is a promising non-invasive technique for assessing specific aspects of the metabolic profile of living mammalian eggs and early embryos, which could be potentially linked to their quality and viability.

Original languageEnglish
Title of host publicationLabel-Free Biomedical Imaging and Sensing, LBIS 2019
PublisherSPIE
Volume10890
ISBN (Electronic)978-1510624221
DOIs
Publication statusPublished - 4 Mar 2019
Externally publishedYes
EventLabel-Free Biomedical Imaging and Sensing 2019, LBIS 2019 - San Francisco, United States
Duration: 2 Feb 20195 Feb 2019

Conference

ConferenceLabel-Free Biomedical Imaging and Sensing 2019, LBIS 2019
Country/TerritoryUnited States
CitySan Francisco
Period2/02/195/02/19

Keywords

  • Coherent raman scattering
  • Lipid metabolism
  • Mammalian eggs and embryos
  • Nonlinear optical microscopy

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