Quantitative imaging of lipids in live mouse oocytes and early embryos using CARS microscopy

Josephine Bradley, Iestyn Pope, Francesco Masia, Randa Sanusi, Wolfgang Langbein, Karl Swann*, Paola Borri

*Awdur cyfatebol y gwaith hwn

Allbwn ymchwil: Cyfraniad at gyfnodolynErthygladolygiad gan gymheiriaid

14 Dyfyniadau (Scopus)
54 Wedi eu Llwytho i Lawr (Pure)

Crynodeb

Mammalian oocytes contain lipid droplets that are a store of fatty acids, whose metabolism plays a substantial role in pre-implantation development. Fluorescent staining has previously been used to image lipid droplets in mammalian oocytes and embryos, but this method is not quantitative and often incompatible with live cell imaging and subsequent development. Here we have applied chemically specific, label-free coherent anti-Stokes Raman scattering (CARS) microscopy to mouse oocytes and preimplantation embryos. We show that CARS imaging can quantify the size, number and spatial distribution of lipid droplets in living mouse oocytes and embryos up to the blastocyst stage. Notably, it can be used in a way that does not compromise oocyte maturation or embryo development. We have also correlated CARS with twophoton fluorescence microscopy simultaneously acquired using fluorescent lipid probes on fixed samples, and found only a partial degree of correlation, depending on the lipid probe, clearly exemplifying the limitation of lipid labelling. In addition, we show that differences in the chemical composition of lipid droplets in living oocytes matured in media supplemented with different saturated and unsaturated fatty acids can be detected using CARS hyperspectral imaging. These results demonstrate that CARS microscopy provides a novel non-invasive method of quantifying lipid content, type and spatial distribution with sub-micron resolution in living mammalian oocytes and embryos.

Iaith wreiddiolSaesneg
Tudalennau (o-i)2238-2247
Nifer y tudalennau10
CyfnodolynDevelopment (Cambridge)
Cyfrol143
Rhif cyhoeddi12
Dynodwyr Gwrthrych Digidol (DOIs)
StatwsCyhoeddwyd - 15 Meh 2016
Cyhoeddwyd yn allanolIe

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