Automated sectioning, imaging, and extraction of renal glomeruli for single-cell analysis Understanding heterogeneous cell states is essential for studying and treating diverse diseases of the kidney. Renal glomeruli, 100-200 ?m across, are abundant but sparse, and implicated in many causes of chronic kidney disease. The proposed method will automate the sectioning, imaging, and extraction of renal glomeruli with single-cell resolution. This approach will sample large populations of glomeruli from human tissue and register serial sections into digital 3d reconstructions. Using this technique, this project aims to ?quantify single cells in whole glomeruli?, ?identify a panel of compatible immunostaining markers, and assess the feasibility of capturing serial frozen tissue sections. ?Together, these will validate new methods to study cell distribution and protein expression in glomerular disease. More broadly, this will facilitate additional automation of sectioning, staining, imaging, and analysis for single-cell experiments. This will enable quantitative, single-cell research in renal pathology, benefiting scientific discovery as well as the development of new treatments.
Project Terms: Algorithms; American; Antigens; Array tomography; Automation; Biopsy Specimen; Cause of Death; Cell Count; cell type; Cells; Chemicals; Chills; Chronic Kidney Failure; clinical application; Computer-Assisted Image Analysis; Development; digital; Dimensions; Disease; Early Diagnosis; Early Intervention; experimental study; Face; Formalin; Freezing; Glass; Goals; Hematoxylin and Eosin Staining Method; Histologic; Histology; Human; human tissue; Image; Image Analysis; Immune; instrument; interest; Kidney; Kidney Diseases; Lung; Mechanics; Methods; Microscope; Microscopic; Morbidity - disease rate; novel strategies; Paraffin Embedding; Pathology; PECAM1 gene; Phase; Population; preservation; Process; protein expression; Proteomics; Protocols documentation; reconstruction; Renal glomerular disease; Renal Tissue; Research; Research Personnel; Resolution; Risk; RNA; sample fixation; Sampling; single cell analysis; Slide; Specimen; Stains; statistics; Structure; Techniques; Technology; Testing; Three-Dimensional Imaging; Tissues; tool; transcriptomics;
