Journal: Investigative Ophthalmology & Visual Science

Article Title: An APE1 Redox Inhibitor Attenuates Pathological Retinal Vascularization by Suppressing FGF2 Angiogenic Signaling

doi: 10.1167/iovs.67.3.47

Figure Lengend Snippet: Refi - 10 reduces pathological neovascularization in oxygen-induced retinopathy (OIR) mice. ( A ) Refi-10 was administered intraperitoneally at P13, and retinas were analyzed at P17. ( B ) Analysis of retinal vasculature. ( a ) Representative IB4-stained whole-mount retinas from normal, OIR, OIR + Refi-10 (25 mg/kg), and OIR + Refi-10 (50 mg/kg) mice at P17. Avascular areas were circled in white . ( b ) Enlarged views of the boxed regions in a . Yellow arrows indicate vascular tufts. ( c ) Neovascularization regions were highlighted in white by ImageJ software. ( d , e ) The quantification of neovascular area and avascular area for each group. Scale bars = 1000 µm. ( C ) Representative H&E staining of retinal sections and quantification of endothelial cell nuclei anterior to the inner limiting membrane. Scale bars = 500 µm. Data are presented as mean ± SD ( n = 6 mice per group; * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001).

Article Snippet: Then, they were incubated with IB4 (1:100, DL-1208-.5; Vector Laboratories, USA) overnight at 4°C.

Techniques: Staining, Software, Membrane

Journal: bioRxiv

Article Title: A cell membrane-hybridized nanocarrier-based RNA delivery system for ocular neovascular diseases

doi: 10.64898/2025.12.23.696136

Figure Lengend Snippet: (a) Fluorescence staining images showing the distribution of liposomes in the choroidal flat-mounts 7 days after intravitreal injection, with neovascular lesions (IB4, green) and liposomes (Cy3, red). (b) Statistical results of fluorescence intensity for each group.

Article Snippet: Griffonia Simplicifolia Lectin I isolectin B4 (IB4) (FL-1201, Fluorescein) was purchased from vector laboratories (USA).

Techniques: Fluorescence, Staining, Liposomes, Injection

Journal: Regenerative Therapy

Article Title: Assessment of permeability in deep tissue capillaries using a new method reflects the nutrient supply status in a healthy heart

doi: 10.1016/j.reth.2025.10.005

Figure Lengend Snippet: Detection of fluorescently labeled dextran leaking from cardiac capillaries A. Three fractionations by fluorescence intensity: background (BG), extra-capillary (BG-BG x a), intra-capillary (>BG x a). B. Methods for calculating background. FITC-conjugated GSL-IB4 was injected into the tail vein 5 min after PBS injection, and another 5 min later, mice were sacrificed, the cardiac tissue was removed, and promptly frozen with isopentane onto dry ice. Frozen tissue sections were cut transversely at 10 μm using the cryostat, and the sections were mounted onto slides. The slides were dried thoroughly and observed under a fluorescence microscope. Twenty 150 μm × 150 μm per 1 section were cut out from the left ventricular wall, where capillaries were observed parallel to the myocardial layer, using LAS X software (Leica), and saved as TIFF (922 x 922 pixels; 22461.544 μm 2 /1 area). Twenty areas per section, three sections per mouse, and three (young) or two (aged) female mice were analyzed, resulting in a total of 180 or 120 images ( see Materials and Methods section for details ). Scale bar: 1 mm. C. Methods for calculating the amount of fluorescently labeled dextran leaked from capillaries. FITC-conjugated GSL-IB4 was injected into the tail vein 5 min after TD 40 injection, and another 5 min later, mice were sacrificed, the cardiac tissue was removed, and promptly frozen with isopentane onto dry ice. Frozen tissue cut sections were prepared in the same manner as above. Fluorescence images of dextran and GSL-IB4 are imported into ImageJ separately, converted to grayscale, and binarized by setting the threshold value for each. The ratio of fluorescence-detected areas to total area was calculated from the acquired fluorescent images. Three sections per mouse, with three mice per month, were analyzed for a total of 180 images. Scale bar: 50 μm.

Article Snippet: To visualize cardiac endothelial cells, fluorescein isothiocyanate (FITC) -conjugated GSL-IB4 (#FL-1201; Vector Laboratories Inc., Burlingame, CA, USA) was injected into the tail vein 5 min after TD 40 injection under anesthesia (1 mg/ml, 50 μl/10 g).

Techniques: Labeling, Fluorescence, Injection, Microscopy, Software