RESUMEN
Assisted reproduction is one of the significant tools to treat human infertility. Morphological assessment is the primary method to determine sperm and embryo viability during in vitro fertilization cycles. It has the advantage of being a quick, convenient, and inexpensive means of assessment. However, visual observation is of limited predictive value for early embryo morphology. It has led many to search for other imaging tools to assess the reproductive potential of a given embryo. The limitations of visual assessment apply to both humans and animals. One recent innovation in assisted reproduction technology imaging is interferometric phase microscopy, also known as holographic microscopy. Interferometric phase microscopy/quantitative phase imaging is the next likely progression of analytical microscopes for the assisted reproduction laboratory. The interferometric phase microscopy system analyzes waves produced by the light as it passes through the specimen observed. The microscope collects the light waves produced and uses the algorithm to create a hologram of the specimen. Recently, interferometric phase microscopy has been combined with quantitative phase imaging, which joins phase contrast microscopy with holographic microscopy. These microscopes collect light waves produced and use the algorithm to create a hologram of the specimen. Unlike other systems, interferometric phase microscopy can provide a quantitative digital image, and it can make 2D and 3D images of the samples. This review summarizes some newer and more promising quantitative phase imaging microscopy systems for evaluating gametes and embryos. Studies clearly show that quantitative phase imaging is superior to bright field microscopy-based evaluation methods when evaluating sperm and oocytes prior to IVF and embryos prior to transfer. However, further assessment of these systems for efficacy, reproducibility, cost-effectiveness, and embryo/gamete safety must take place before they are widely adopted.
Asunto(s)
Embrión de Mamíferos , Holografía , Holografía/métodos , Animales , Humanos , Embrión de Mamíferos/diagnóstico por imagen , Embrión de Mamíferos/fisiología , Masculino , Femenino , Células Germinativas/fisiología , Espermatozoides/fisiología , Técnicas Reproductivas Asistidas , Fertilización In Vitro/métodos , Microscopía/métodos , Microscopía/instrumentaciónRESUMEN
The surgical pathology workflow currently adopted by clinics uses staining to reveal tissue architecture within thin sections. A trained pathologist then conducts a visual examination of these slices and, since the investigation is based on an empirical assessment, a certain amount of subjectivity is unavoidable. Furthermore, the reliance on external contrast agents such as hematoxylin and eosin (H&E), albeit being well-established methods, makes it difficult to standardize color balance, staining strength, and imaging conditions, hindering automated computational analysis. In response to these challenges, we applied spatial light interference microscopy (SLIM), a label-free method that generates contrast based on intrinsic tissue refractive index signatures. Thus, we reduce human bias and make imaging data comparable across instruments and clinics. We applied a mask R-CNN deep learning algorithm to the SLIM data to achieve an automated colorectal cancer screening procedure, i.e., classifying normal vs. cancerous specimens. Our results, obtained on a tissue microarray consisting of specimens from 132 patients, resulted in 91% accuracy for gland detection, 99.71% accuracy in gland-level classification, and 97% accuracy in core-level classification. A SLIM tissue scanner accompanied by an application-specific deep learning algorithm may become a valuable clinical tool, enabling faster and more accurate assessments by pathologists.
Asunto(s)
Neoplasias Colorrectales , Aprendizaje Profundo , Neoplasias Colorrectales/diagnóstico , Detección Precoz del Cáncer , Humanos , Microscopía , Microscopía de Interferencia/métodosRESUMEN
Characterization of spermatozoon viability is a common test in treating infertility. Recently, it has been shown that label-free, phase-sensitive imaging can provide a valuable alternative for this type of assay. We employ spatial light interference microscopy (SLIM) to perform high-accuracy single-cell phase imaging and decouple the average thickness and refractive index information for the population. This procedure was enabled by quantitative-phase imaging cells on media of two different refractive indices and using a numerical tool to remove the curvature from the cell tails. This way, we achieved ensemble averaging of topography and refractometry of 100 cells in each of the two groups. The results show that the thickness profile of the cell tail goes down to 150 nm and the refractive index can reach values of 1.6 close to the head.