3D time-lapse imaging of a mouse embryo using intensity diffraction tomography embedded inside a deep learning...

Area of Science:

• Biophotonics
• Developmental Biology
• Microscopy

Background:

• Long-term live imaging of developing organisms is crucial for understanding biological processes.
• Existing microscopy techniques often require complex setups or labels, limiting their applicability for extended observations.

Purpose of the Study:

• To present a compact, robust, and label-free 3D diffractive microscope for long-term imaging of developing organisms within cell incubators.
• To reconstruct the 3D optical properties (refractive index and absorption) of large specimens with subcellular resolution.

Main Methods:

• Utilized a compact 3D diffractive microscope with no moving parts, compatible with standard cell culture containers.
• Employed an LED array for multi-angle illumination and reconstructed 3D optical properties using a deep neural network trained with the beam propagation method.
• Accounted for spherical aberrations from interfaces in the forward model.

Main Results:

• Successfully performed time-lapse 3D imaging of preimplantation mouse embryos over six days, from single-cell to blastocyst stages.
• Achieved subcellular resolution, enabling quantitative assessment of embryo development and viability.
• Demonstrated the system's capability in both low-scattering and highly scattering regimes.

Conclusions:

• The developed 3D diffractive microscope offers a novel, label-free approach for long-term live imaging of whole embryos and organoids.
• This technology provides new opportunities for quantitative analysis of biological development and viability.
• The system's simplicity, robustness, and compatibility with incubators make it suitable for extended developmental studies.