surface surface coloc xtension software  (Oxford Instruments)

Journal: Advanced Materials (Deerfield Beach, Fla.)

Article Title: Microgels With Electrostatically Controlled Molecular Affinity to Direct Morphogenesis

doi: 10.1002/adma.202409731

Figure Lengend Snippet: µGUIDEs as engineered signaling centers to locally stimulate kidney organoid vascularization. a,b), Colocalization of EC networks and emerging epithelial structures in human iPSC‐based kidney organoid cultures. a), Day 7 immunofluorescence staining showing PAX2+ RVs and CD31+ ECs. Scale bars: 1 mm (left) and 500 µm (right). b), Day 17 Immunofluorescence staining highlighting WT1+ glomeruli, LTL+ proximal tubule cells, and CD31+ ECs. Scale bars: 1 mm (left), 500 µm (right). c), Schematic representation of the µGUIDE deposition within kidney organoids. d), 3D simulation of the kidney organoid transwell culture depicting the VEGF gradient formation from starPEG‐sGAG µGUIDEs using a reaction‐diffusion model. The simulations show the concentration profiles of freely diffusing VEGF (normalized by initial µGUIDE loading) between two adjacent µGUIDEs, 100 µm above the organoid base (dashed line in the schematic on the top left). The µGUIDE integral charge density (P1) is concluded to determine the duration of the VEGF gradient, from short‐term (hours) to long‐term (days). e), Phase contrast images of kidney organoids with P1 5 P2 3.6 µGUIDEs at day 0 (left; scale bar: 1 mm) and day 7 (right; scale bar: 200 µm). The arrowhead in the right image indicates a µGUIDE. f), Day 7 confocal images of kidney organoids cultured with VEGF‐loaded µGUIDEs (w/ VEGF), unloaded µGUIDEs (w/o VEGF) and no µGUIDEs (w/o µGUIDE). Scale bars, 1 mm. g), Quantitative analysis of the EC response (CD31 coverage area) to VEGF gradients around and between µGUIDEs at day 7. Control regions of interest (ROIs) of identical size were positioned in organoids without µGUIDEs. h,i), Analysis of the interactions between RVs and ECs. h), Confocal images of day 7 organoids stained for PAX2+ RVs and CD31+ ECs, with magenta indicating the contact area. Scale bar, 500 µm. i), Quantitative analysis of the RV‐EC contact. For all experiments, µGUIDEs with a diameter of ≈ 182 µm were used. Data are shown as mean ± s.d. ( n = 4).

Article Snippet: Here, two surfaces were created for the respective source channels, and the surface‐surface colocalization Imaris XTension was applied.

Techniques: Immunofluorescence, Staining, Diffusion-based Assay, Concentration Assay, Cell Culture, Control