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First ever “5k” ultra-HD highest resolution video imaging of gametes and embryos: pilot study for morphological assessment

2016; Elsevier BV; Volume: 106; Issue: 3 Linguagem: Inglês

10.1016/j.fertnstert.2016.07.646

1556-5653

S.F. Palter, Marcos Meseguer, Joana Santos, J.L. Romero, António Pellicer,

Sperm and Testicular Function

Morphology is the gold-standard method for assessment of gametes and embryos. Despite this, imaging resolution has not changed significantly for many years. Visualization requires either direct visualization through microscope objectives (limited magnification) or image capture via CCD/CMOS chip and inspection on a monitor (limited resolution). Advances in digital imaging technology have recently created ultra-high definition (“4k” and “5k”) systems which allow 4-10X greater resolution of images versus standard definition imaging. Until now, it has not been possible to use these systems for microscopy. We report the development of the first 5k microscopy imaging system and the first pilot study of its utility for assessment of gamete and embryo morphology. Prospective observational study. A special imaging system was designed for digital microscopy imaging using the highest resolution next generation digital video (“5K” video at 5120 × 2700 resolution; Scarlet Dragon, Red Digital Cinema Camera Company). A custom designed coupling system allowed imaging through an inverted microscope. 860 recordings of consecutive cases were made during IVF of sperm, oocytes, ICSI procedures, zygotes, cleavage stage embryos, and blastocysts at 400x-960x magnification. Images were viewed and scored in on a 55 inch 4k ultraHD monitor providing additional magnification factors. Specialized hardware/software allowed real-time full motion full resolution viewing of videos. High frame rate capture (96 fps) was used to digitally slow the movement of individual sperm. All videos were reviewed by two embryologists. The system was successfully able to image gametes and embryos at 5k ultra HD. Images viewed at full resolution/high magnification provided enhanced structural clarity and detail. See Table for results by cell type.Tabled 1ResultsStructureImage QualitySpermMotile sperm morphology is comparable to IMSI. High frame rate recording digitally slowed motion increasing morphologic detail and simplifying selectionOocyteUltrastructural analysis superior to standard microscopy including differentiation of zona pellucida, cytoplasm and polar body. No new structures were identified.ZygoteVery detailed PN with increased definition of symmetry and nucleoli. Increased definition of polar bodies and small fragments. Increased definition of morphological alterations including cytoplasm intrusions, periviteline space, zona pellucida, cytoplasmic halo, and vacuoles , and halo.Clevage EmbryoImage resolution superior to standard microscopy with increased visualization of nuclei, multinucleation, the zona as a sphere, vacuoles and granularity of cytoplasm. On compacted morula cell limits are well-defined.BlastocystSuperior visualization and all individual cells of trophectoderm and ICM are easily delineated. On expanded blastocysts, a novel observation noted was nuclei from all blastomeres were visible. Open table in a new tab It is possible to video image living human gametes and embryos at extreme magnification in ultra high definition “5K” resolution. Cellular structural detail and image quality surpassed that of standard methods and could replace traditional microscopy. Images can be digitally slowed to arrest the motion of living sperm, potentially improving sperm selection at ICSI. For eggs and embryos, resolution and magnification did not identify new morphological features. However, the ability to visualize and quantify individual cells, intracellular structures, and nucleation in blastocysts could be a novel mechanism for embryo assessment. Further validation studies are underway.