Lensfree diffractive tomography for the imaging of 3D cell cultures

International audienceNew microscopes are needed to help realize the full potential of 3D organoid culture studies. In order to image large volume of 3D organoid cultures while preserving the ability to catch every single cell, we propose a new imaging platform based on lensfree microscopy. We have built a lensfree diffractive tomography setup performing multi-angle acquisitions of 3D organoid culture embedded in Matrigel ® and developed a dedicated 3D holographic reconstruction algorithm based on the Fourier diffraction theorem. With this new imaging platform, we have been able to reconstruct a 3D volume as large as 21.5 mm 3 of a 3D organoid culture of prostatic RWPE1 cells showing the ability of these cells to assemble in 3D intricate cellular network at the mesoscopic scale. Importantly, comparisons with 2D images show that it is possible to resolve single cells isolated from the main cellular structure with our lensfree diffractive tomography setup

A dynamical measure of the black hole mass in a quasar 11 billion years ago

Tight relationships exist in the local universe between the central stellar properties of galaxies and the mass of their supermassive black hole. These suggest galaxies and black holes co-evolve, with the main regulation mechanism being energetic feedback from accretion onto the black hole during its quasar phase. A crucial question is how the relationship between black holes and galaxies evolves with time; a key epoch to probe this relationship is at the peaks of star formation and black hole growth 8-12 billion years ago (redshifts 1-3). Here we report a dynamical measurement of the mass of the black hole in a luminous quasar at a redshift of 2, with a look back time of 11 billion years, by spatially resolving the broad line region. We detect a 40 micro-arcsecond (0.31 pc) spatial offset between the red and blue photocenters of the H$\alpha$ line that traces the velocity gradient of a rotating broad line region. The flux and differential phase spectra are well reproduced by a thick, moderately inclined disk of gas clouds within the sphere of influence of a central black hole with a mass of 3.2x10$^{8}$ solar masses. Molecular gas data reveal a dynamical mass for the host galaxy of 6x10$^{11}$ solar masses, which indicates an under-massive black hole accreting at a super-Eddington rate. This suggests a host galaxy that grew faster than the supermassive black hole, indicating a delay between galaxy and black hole formation for some systems.Comment: 5 pages Main text, 8 figures, 2 tables, to be published in Nature, under embargo until 29 January 2024 16:00 (London

3D LENSFREE MICROSCOPY FOR 3D CELL CULTURE

International audienceNew microscopes are needed to help realize the full potential of 3D organoid culture studies by gathering large quantitative and systematic data over extended period of time while preserving the integrity of the living sample. In order to reconstruct large volume while keeping the ability to catch every single cell, we propose new imaging platforms based on lensfree microscopy, a technic which is addressing these needs in the context of 2D cell culture, providing label-free and non-phototoxic acquisition of large datasets. We have built lensfree diffractive tomography setups performing multi-angle acquisitions of 3D organoid culture embedded in Matrigel ® and developed dedicated 3D holographic reconstruction algorithms based on the Fourier diffraction theorem. Nonetheless, holographic setups do not record the phase of the incident wavefront and the biological samples in Petri dish strongly limit the angular coverage. These limitations introduces numerous artefacts in the sample reconstruction. We developed several methods to overcome them, such as multi wavelength imaging or iterative phase retrieval. The most promising technic currently developed is based on a regularized inverse problem approach directly performed on the 3D volume to reconstruct. 3D reconstructions were realized on several complex samples such as 3D networks or spheroids embedded in capsules with large reconstructed volumes up to ~25 mm^3 while still being able to identify single cells. To our knowledge, this is the first time that such an inverse problem approach is implemented in the context of lensfree diffractive tomography enabling to reconstruct large volume of unstained biological samples

Comparative study of fully three-dimensional reconstruction algorithms for lens-free microscopy

International audienceWe propose a three-dimensional (3D) imaging platform based on lens-free microscopy to perform multiangle acquisitions on 3D cell cultures embedded in extracellular matrices. Lens-free microscopy acquisitions present some inherent issues such as the lack of phase information on the sensor plane and a limited angular coverage. We developed and compared three different algorithms based on the Fourier diffraction theorem to obtain fully 3D reconstructions. These algorithms present an increasing complexity associated with a better reconstruction quality. Two of them are based on a regularized inverse problem approach. To compare the reconstruction methods in terms of artefact reduction, signal-to-noise ratio, and computation time, we tested them on two experimental datasets: an endothelial cell culture and a prostate cell culture grown in a 3D extracellular matrix with large reconstructed volumes up to ∼5  mm3∼5  mm3 with a resolution sufficient to resolve isolated single cells. The lens-free reconstructions compare well with standard microscopy

Estimation of the lateral mis-registrations of the GRAVITY + adaptive optics system: Perturbative method with open-loop modal correlation and non-perturbative method with temporal correlation of closed-loop telemetry

International audienceContext. The GRAVITY+ upgrade implies a complete renewal of its adaptive optics (AO) systems. Its complex design, featuring moving components between the deformable mirrors and the wavefront sensors, requires the monitoring and auto-calibrating of the lateral mis-registrations of the system while in operation. Aims. For preset and target acquisition, large lateral registration errors must be assessed in open loop to bring the system to a state where the AO loop closes. In closed loop, these errors must be monitored and corrected, without impacting the science. Methods. With respect to the first requirement, our method is perturbative, with two-dimensional modes intentionally applied to the system and correlated to a reference interaction matrix. For the second requirement, we applied a non-perturbative approach that searches for specific patterns in temporal correlations in the closed loop telemetry. This signal is produced by the noise propagation through the AO loop. Results. Our methods were validated through simulations and on the GRAVITY+ development bench. The first method robustly estimates the lateral mis-registrations, in a single fit and with a sub-subaperture resolution while in an open loop. The second method is not absolute, but it does successfully bring the system towards a negligible mis-registration error, with a limited turbulence bias. Both methods proved to robustly work on a system still under development and not fully characterised. Conclusions. Tested with Shack-Hartmann wavefront sensors, the proposed methods are versatile and easily adaptable to other AO instruments, such as the pyramid, which stands as a baseline for all future AO systems. The non-perturbative method, not relying on an interaction matrix model and being sparse in the Fourier domain, is particularly suitable to the next generation of AO systems for extremely large telescopes that will present an unprecedented level of complexity and numbers of actuators