![]() ![]() ![]() They are particularly suitable for phenotypic screening studies at the tissue- and organ specific level. ![]() Importantly, the tools are generic by design and were benchmarked on several microscopy datasets of zebrafish larvae and medaka embryos. The resulting software tools address various steps of the screening workflow, from manual ground-truth annotations, automated detection of regions of interest, targeted imaging of specific tissues or organs using feedback microscopy, image-classification, and interactive data exploration. The aim was to create versatile and robust solutions tailored to the requirements of microscopy-based phenotypic screening studies in small model organisms. This dissertation reports the design, development and benchmarking of novel research software inspired by the field of computer vision and data-science. The present findings contribute to better understanding of the factors affecting the topography of retinal ganglion cells and mechanisms of visual adaptation in fish. japonicus are consistent with its highly visual behaviour. The relatively high density of GCs and presence of the horizontal streak and area retinae temporalis in the H. The theoretical anatomical spatial resolution (the anatomical estimate of the upper limit of visual acuity) was minimum in the ventral periphery (smaller fish, 1.43 cpd larger fish, 1.37 cpd) and maximum in area retinae temporalis (smaller fish, 2.83 cpd larger fish, 2.41 cpd). The maximum cell density (area retinae temporalis) ranged from 9492 to 14,112 cells/mm2, and the total number of GCs varied from 286 x 103 to 326 x 103 cells in three individuals. Two zones of increased GC density in the nasal and temporal retina were bridged by a horizontal streak with the GC density ranging from 5600 to 8000 cells/mm2. DAPI labelling was used to visualize cell nuclei in the ganglion cell and inner plexiform layers. The eyes and retinae were examined by light microscopy and computerized tomography. The present study deals with the topography of retinal ganglion cells (GCs) and spatial resolution in the smelt Hypomesus japonicus. This enhanced and novel method is clinically significant as it could be utilized in the future to more efficiently produce mature, secondary oocytes, for use with IVF/ICSI to treat POF patients. During in vitro culture, isolated cells retained the germline phenotype expression of DDX4 and IFITM3 as confirmed by gene expression analysis, demonstrated characteristic germline stem cell self-assembly into embryoid bodies, and formed > 40 µm “oocyte-like” structures that expressed the early oocyte markers GDF9, DAZL, and ZP1. MACS sorting ovarian cells using a the two-marker combination yielded a ~ twofold higher percentage of OSCs from a given mass of ovarian tissue compared to existing single marker methods while minimizing the debate surrounding germline marker selection. Our study supports earlier findings of OSCs in ovaries during the entire female lifespan: from reproductive age through post-menopausal age. ![]() We hypothesized that by using anti-DDX4 and anti-IFITM3 antibodies, in combination, with MACS, we would improve the yield of isolated OSCs versus using anti-DDX4 antibodies alone. Separate groups, however, have efficiently isolated OSCs using another antibody marker Ifitm3 which is consistently recognized to be transmembrane in location. Previously, the DDX4 antibody marker alone was utilized to isolate OSCs however, extensive debate over its location in OSCs versus resulting oocytes (transmembrane or intracytoplasmic) has raised doubt about the identity of these cells. Oogonial stem cells (OSCs), found in adult ovaries, have provided insight into potential paths to treating infertility. Alternative methods to obtain mature oocytes are still needed for women with premature ovarian failure (POF). ![]()
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