Journal: bioRxiv
Article Title: Combined TIRF and 3D Super-Resolution Microscopy for Nanoscopic Spatiotemporal Characterization of Adhesion Molecules on Microvilli
doi: 10.1101/2025.08.26.672391
Figure Lengend Snippet: Surface topography of KG1a cells reconstructed by TIRF microscopy. (a, b) Fluorescence images of KG1a cells stained by (a) MemBrite FX640 and (b) DiD-Vybrant membrane stains, captured using wide-field epi-illumination microscopy. (c) Fluorescence image of the bottom surface of the MemBrite FX640-stained KG1a cell, captured by TIRFM. (d) Filtered image of (c) using Laplacian of Gaussian (LoG) filter; Bottom view (top panel) and side view (bottom panel). The color bar indicates fluorescence intensity values. (e) 3D topographic map reconstructed from the fluorescence image shown in (d) using ; Bottom view (top panel) and side view (bottom panel). The color bar represents the distance from the glass surface. (f) Contours of individual microvilli in the topographic map shown in (e), obtained using an edge detection algorithm applied to the binary mask generated by thresholding the image. (g) Enlarged views of the regions highlighted by the rectangles in (f). δz values are red color-coded, with a step size of 10 nm. (h) SEM image of a KG1a cell, highlighting microvilli structures. (i) Comparative analysis of the average number of microvilli per µm² detected using SEM and TIRFM.
Article Snippet: KG1a cells, a human acute myelogenous leukemia cell line (ATCC), were maintained in RPMI (Gibco) supplemented with 10% fetal bovine serum, penicillin (100 U ml -1 ), and streptomycin (100 μg ml -1 ).
Techniques: Microscopy, Fluorescence, Staining, Membrane, Generated