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Journal: iScience
Article Title: Serial intravital microscopy reveals temporal dynamics of autoreactive germinal centers in the spleen
doi: 10.1016/j.isci.2026.115340
Figure Lengend Snippet: An AIW to track FDC network dynamics in the spleen over 2 weeks using serial intravital microscopy (A) Graphical overview of surgery steps of AIW (abdominal imaging window) implantation: 1. surgical landmarks, 2. purse string suture, 3. spleen mobilization, 4. placement of AIW, and 5. recovered mouse after surgery. (B) Recovered mouse 8 days after AIW implantation. (C) Global treatment timeline: Foxp3 DTR−GFP mice were treated with R848 three times per week for four weeks to induce an autoimmune response. An AIW was implanted either shortly after R848 treatment initiation or after 4 weeks of treatment. GCs were then followed on six days over 15 days total during or after treatment, respectively. To visualize FDCs in the light zone, mice received αCD35-iFluor647 i.v. 24 h before every imaging session. (D) Maximum intensity projection of a 3D-stitched overview z-stack (100–250 μm below capsule, 150 μm thick) of the entire AIW FOV (8×10 tiles). Asterisks indicate the most superficial FDC networks. White arrowheads point to a large blood vessel, used as a landmark for reorientation over time. Green arrows indicate the borders of AIW FOV. Scale bar, 500 μm. Image was processed with background subtraction, median filtering (Despeckle), linear brightness, and contrast adjustment. See also and .
Article Snippet:
Techniques: Intravital Microscopy, Imaging
Journal: iScience
Article Title: Serial intravital microscopy reveals temporal dynamics of autoreactive germinal centers in the spleen
doi: 10.1016/j.isci.2026.115340
Figure Lengend Snippet: Processing of deep 3D data to enhance single-cell resolution, and spatiotemporal tracking using PA-GFP (A–C) Imaging frames of the same FOV 75, 145, and 185 μm below the capsule (dual track overlay from λ Ex 840 and λ Ex 940). FoxP3-GFP + cells are mostly visible, but with a low signal-to-noise ratio (SNR). Scale bars, 100 μm. (D–F) Same images as A-C, after background subtraction and median filtering (Despeckle). (G–I) Same images as D-F, after 3D Maximum filter. White arrow identifies a FoxP3 + cell. Scale bars, 100 μm, calculated by pixels as precise scaling is not possible after 3D maxima filtering due to the spatial enlargement of single spots. (J) Longitudinal tracking of PA-GFP-stimulated area (collagen trabecula) for 2 weeks. Scale bars, 100 μm. Experimental n = 3. All micrographs were adjusted in brightness and contrast (linear); any further processing is indicated in panel legends above.
Article Snippet:
Techniques: Single Cell, Imaging