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A decrease in neuronal complexity was seen in hippocampus of 10 month old Arctic mice at the time that correlates with the behavior deficit, both of which were rescued in the Arctic/C5aR1KO. In addition, phagosomal-lysosomal gene expression was increased in the Arctic mice relative to wild type but further increased in the Arctic/C5aR1KO mice.
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RNA-seq analysis identified inflammation related genes as differentially expressed, with increased expression in the Arctic mice relative to wild type and decreased expression in the Arctic/C5aR1KO relative to Arctic. Microglia were sorted from infiltrating monocytes (GFP and RFP-positive) for transcriptome analysis. Immunohistochemical analysis showed no CCR2 + monocytes/macrophages near the plaques in the Arctic brain with or without C5aR1. ResultsĪ lack of C5aR1 prevented behavior deficits at 10 months, although amyloid plaque load was not altered. CX3CR1 GFP and CCR2 RFP reporter mice were bred to C5aR1 sufficient and knockout wild type and Arctic mice to enable sorting of microglia (GFP-positive, RFP-negative) isolated from adult brain at 2, 5, 7 and 10 months of age followed by RNA-seq analysis. MethodsĬ5aR1 knock out mice were crossed to the Arctic AD mouse model, and characterized for pathology and for behavior performance in a hippocampal dependent memory task. In addition, since C5aR1 is primarily expressed on cells of the myeloid lineage, and only to a lesser extent on endothelial cells and neurons in brain, gene expression in microglia isolated from adult brain at multiple ages was compared across all genotypes. To validate that the effect of the antagonist was specifically via C5aR1 inhibition, mice lacking C5aR1 were generated and compared in behavior and pathology.
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Microtome (Microm HM 355S, Thermo Scientific).Aperio ScanScope FL (Fluorescent) for whole slide immunohistological tissue section scanning up to 40X magnification (Aperio Inc.Aperio ScanScope CS (for Bright Field) for whole histological tissue section scanning up to 40X magnification (Aperio Inc.Zeiss upright Microscope for both Brightfield and Fluorescent experiments (Carl Zeiss, Germany).
STEREOLOGY IHC AXIO SOFTWARE
Zeiss Imager Z1 with Microbrightfield hardware and software (MBF, USA) for morphometric and stereological analyses of immuno-histological samples (Carl Zeiss, Germany).Zeiss Imager Z2 with Axio Vision software (Carl Zeiss, Germany).Zeiss single photon Confocal Laser Microscope LSM710 with ZEN software (Carl Zeiss, Germany).Zeiss Multiphoton Confocal Laser Microscope LSM880 with Airyscan (Carl Zeiss, Germany).This list of equipment is used for qualitative and quantitative analysis of the distribution, co-localization and co-expression of various neurotransmitters/neuromodulators in different regions of the brain and spinal cord sections including stereological studies. Histology, Immunohistochemistry and Microscopy Show submenu for Neurophysiological Outcomes Histology, Immunohistochemistry, Microscopic Experiments Cellular and Molecular Outcomes Overview.current page Show submenu for Cellular and Molecular Outcomes Rodent Post-Surgery and Critical Care Monitoring Equipment