Optical matrix imaging applied to embryology

High-resolution label-free imaging of oocytes and embryos is essential for in vitro fertilization procedures. Yet conventional microscopy fails in this task because of aberrations and multiple scattering induced by refractive index heterogeneities inside the sample. These detrimental phenomena drastically degrade the images of early embryos particularly in depth. To overcome these fundamental problems without sacrificing the frame rate, optical matrix imaging (OMI) is a suitable tool. Relying on an ultra-fast measurement of the reflection matrix associated with the sample, it can compensate for aberration and forward multiple scattering in post-processing, thereby providing three-dimensional and highly contrasted images of embryos at a confocal resolution. As a first proof-of-concept, bovine oocytes and embryos are imaged at a 300 nm resolution almost in real time. Our system enables visualization of intracellular structures such as lipids and mitochondria in the cytoplasm or the zona pellucida surrounding it. Altogether, we demonstrate that OMI is a promising tool for research in developmental biology and for time-lapse monitoring of oocytes and embryos in assisted reproduction.

Multi-Spectral Reflection Matrix for Ultra-Fast 3D Label-Free Microscopy

Label-free microscopy exploits light scattering to obtain a three-dimensional image of biological tissues. However, light propagation is affected by aberrations and multiple scattering, which drastically degrade the image quality and limit the penetration depth. Multi-conjugate adaptive optics and time-gated matrix approaches have been developed to compensate for aberrations but the associated frame rate is extremely limited for 3D imaging. Here we develop a multi-spectral matrix approach to solve these fundamental problems. Based on an interferometric measurement of a polychromatic reflection matrix, the focusing process can be optimized in post-processing at any voxel by addressing independently each frequency component of the wave-field. A proof-of-concept experiment demonstrates the three-dimensional image of an opaque human cornea over a 0.1 mm^3-field-of-view at a 290 nm-resolution and a 1 Hz-frame rate. This work paves the way towards a fully-digital microscope allowing real-time, in-vivo, quantitative and deep inspection of tissues.