anti nephrin antibody  (R&D Systems)

Journal: medRxiv

Article Title: Nanoscale Podocyte Morphometrics Predict Disease Progression in IgA Nephropathy

doi: 10.64898/2026.03.30.26349728

Figure Lengend Snippet: (a) OCT frozen kidney biopsies were sectioned, cleared and immunolabelled followed by confocal imaging and deep-learning segmentation. (b) Maximum intensity projection confocal image of nephrin visualizing the slit diaphragm network and foot processes. (c) Output of the semantic segmentation of both the slit diaphragm and foot processes. Foot processes are marked in red and the slit diaphragm in green. (d) Output of the instance segmentation resulting in segmentation of each individual foot process in the image. Each foot process is presented in an individual color. (e-g) From the 2 segmentation outputs, the morphometric parameters SDL (e), FP circularity (f), FP area and FP perimeter (g) are extracted. (e) Schematic image of how SDL is measured. The length of the slit (in blue) is divided by the surface area (outlined in white) resulting in a length-density measurement of slit coverage. (f) Schematic image of how FP circularity is measured. From the segmentation of the individual foot processes, the FP circularity is calculated using the formula, circularity = 4 π (area/perimeter−2). The result is a value between 0 and 1 where a perfect circle has a circularity of 1 and a very elongated rectangle approaches 0. (g) Foot process size is calculated both by FP area and by FP perimeter. The FP area is the pixel area of each segmented foot process converted to µm 2 and the FP perimeter is the outline of the foot process with it closed at the base.

Article Snippet: OCT-frozen biopsy specimens were fixed in PFA, cleared in 4% SDS w/ boric acid and fluorescently immunolabelled for nephrin (R&D Systems, cat. no. AF4269) to visualize the slit diaphragm and podocyte foot processes (FPs) based on the protocol described by Unnersjö-Jess et al (details available in Supplement Methods).

Techniques: Imaging