RESUMEN
Embryo quality is an important determinant of successful implantation and a resultant live birth. Current clinical approaches for evaluating embryo quality rely on subjective morphology assessments or an invasive biopsy for genetic testing. However, both approaches can be inherently inaccurate and crucially, fail to improve the live birth rate following the transfer of in vitro produced embryos. Optical imaging offers a potential non-invasive and accurate avenue for assessing embryo viability. Recent advances in various label-free optical imaging approaches have garnered increased interest in the field of reproductive biology due to their ability to rapidly capture images at high resolution, delivering both morphological and molecular information. This burgeoning field holds immense potential for further development, with profound implications for clinical translation. Here, our review aims to: (1) describe the principles of various imaging systems, distinguishing between approaches that capture morphological and molecular information, (2) highlight the recent application of these technologies in the field of reproductive biology, and (3) assess their respective merits and limitations concerning the capacity to evaluate embryo quality. Additionally, the review summarizes challenges in the translation of optical imaging systems into routine clinical practice, providing recommendations for their future development. Finally, we identify suitable imaging approaches for interrogating the mechanisms underpinning successful embryo development.
Asunto(s)
Imagen Óptica , Humanos , Imagen Óptica/métodos , Animales , Desarrollo Embrionario/fisiología , Embrión de Mamíferos/diagnóstico por imagen , Embrión de Mamíferos/fisiología , Femenino , EmbarazoRESUMEN
Intensity shot noise in digital holograms distorts the quality of the phase images after phase retrieval, limiting the usefulness of quantitative phase microscopy (QPM) systems in long term live cell imaging. In this paper, we devise a hologram-to-hologram neural network, Holo-UNet, that restores high quality digital holograms under high shot noise conditions (sub-mW/cm2 intensities) at high acquisition rates (sub-milliseconds). In comparison to current phase recovery methods, Holo-UNet denoises the recorded hologram, and so prevents shot noise from propagating through the phase retrieval step that in turn adversely affects phase and intensity images. Holo-UNet was tested on 2 independent QPM systems without any adjustment to the hardware setting. In both cases, Holo-UNet outperformed existing phase recovery and block-matching techniques by â¼ 1.8 folds in phase fidelity as measured by SSIM. Holo-UNet is immediately applicable to a wide range of other high-speed interferometric phase imaging techniques. The network paves the way towards the expansion of high-speed low light QPM biological imaging with minimal dependence on hardware constraints.
RESUMEN
Back focal plane interferometry (BFPI) is one of the most straightforward and powerful methods for achieving sub-nanometer particle tracking precision at high speed (MHz). BFPI faces technical challenges that prohibit tunable expansion of linear detection range with minimal loss to sensitivity, while maintaining robustness against optical aberrations. In this paper, we devise a tunable BFPI combining a structured beam (conical wavefront) and structured detection (annular quadrant photodiode). This technique, which we termed Structured Back Focal Plane Interferometry (SBFPI), possesses three key novelties namely: extended tracking range, low loss in sensitivity, and resilience to spatial aberrations. Most importantly, the conical wavefront beam preserves the axial Gouy phase shift and lateral beam waist that can then be harnessed in a conventional BFPI system. Through a series of experimental results, we were able to tune detection sensitivity and detection range over the SBFPI parameter space. We also identified a figure of merit based on the experimental optimum that allows us to identify optimal SBPFI configurations that balance both range and sensitivity. In addition, we also studied the resilience of SBFPI against asymmetric spatial aberrations (astigmatism of up to 0.8 λ) along the lateral directions. The simplicity and elegance of SBFPI will accelerate its dissemination to many associated fields in optical detection, interferometry and force spectroscopy.
RESUMEN
The ability to label individual cells is useful for single-cell-level studies of complex cellular interactions and heterogeneity. Optically readable cell labeling is attractive as it can be investigated non-invasively and repeatedly at high speeds. Here, we demonstrate the feasibility of large-scale cell barcoding and identification using fluorescent polystyrene microbeads loaded into cells. Intracellular beads with different diameters in a range of 5 to 12 µm generate spectrally distinguished features or barcodes. A microfluidic chip was used to measure fluorescence resonance peaks emitted from individual cells. An algorithm comparing the peak wavelengths to a reference barcode library allowed barcode identification with high accuracy. This work provides a guideline to increase the number of unique identifiers and reduce various false-positive and false-negative errors.
Asunto(s)
Procesamiento Automatizado de Datos , Técnicas Analíticas Microfluídicas/instrumentación , Análisis de la Célula Individual/instrumentación , Análisis de la Célula Individual/métodos , Algoritmos , Colorantes Fluorescentes/química , Microscopía Fluorescente , Microesferas , PoliestirenosRESUMEN
Efficient delivery of viruses, proteins and biological macromelecules into a micrometer-sized focal spot of an XFEL beam for coherent diffraction imaging inspired new development in touch-free particle injection methods in gaseous and vacuum environments. This paper lays out our ongoing effort in constructing an all-optical particle delivery approach that uses piconewton photophoretic and femtonewton light-pressure forces to control particle delivery into the XFEL beam. We combine a spatial light modulator (SLM) and an electrically tunable lens (ETL) to construct a variable-divergence vortex beam providing dynamic and stable positioning of levitated micrometer-size particles, under normal atmospheric pressure. A sensorless wavefront correction approach is used to reduce optical aberrations to generate a high quality vortex beam for particle manipulation. As a proof of concept, stable manipulation of optically-controlled axial motion of trapped particles is demonstrated with a response time of 100ms. In addition, modulation of trapping intensity provides a measure of the mass of a single, isolated particle. The driving signal of this oscillatory motion can potentially be phase-locked to an external timing signal enabling synchronization of particle delivery into the x-ray focus with XFEL pulse train.