Journal: Physiological Reports

Article Title: Autologous minced muscle grafts improve endogenous fracture healing and muscle strength after musculoskeletal trauma

doi: 10.14814/phy2.13362

Figure Lengend Snippet: Flow cyotometry antibodies

Article Snippet: CD68 , ED1 , Novus Biologicals , NB600‐985V , 0.001 mg/mL.

Techniques: Concentration Assay

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Targeted muscle reinnervation attenuates neuropathic pain and neuroma development in a rat model of tibial nerve transection

doi: 10.3389/fbioe.2026.1758496

Figure Lengend Snippet: Immunofluorescence evaluation of the distal segment of the tibial nerve. (A) Immunofluorescence expression of CGRP and TRPA1 in longitudinal sections of the distal tibial nerve from each experimental group (scale bar = 100 µm). (B,C) Quantitative analysis of the immunopositive areas for CGRP and TRPA1. (D) Immunofluorescence expression of CD3, and CD68 in longitudinal sections of the distal tibial nerve from each experimental group (scale bar = 100 µm). (E,F) Statistical analysis of T lymphocytes (CD3) and macrophages (CD68) in each experimental group. Data are expressed as means ± SD, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001, ns: no significance. n = 5 for all groups.

Article Snippet: Nuclei were counterstained with DAPI-containing mounting medium (Bioss, C02-04002). (2) CD3/CD68 (tibial nerve sections, on separate sections from CGRP/TRPA1): anti-CD3 antibody (Mouse, 1:200, Bioss, bsm-54036M) and anti-CD68 antibody (Rabbit, 1:200, Bioss, bs-1432R).

Techniques: Immunofluorescence, Expressing

Journal: Journal of vascular and interventional radiology : JVIR

Article Title: MR Imaging–Based In Vivo Macrophage Imaging to Monitor Immune Response after Radiofrequency Ablation of the Liver

doi: 10.1016/j.jvir.2022.11.013

Figure Lengend Snippet: An illustrative representation of the experimental design for magnetic resonance imaging of immune cells in vivo. (a) Radiofrequency ablation of mouse normal liver, which allowed the establishment of an inflamed periablational rim for 7 days after ablation. Two separate groups of 12 mice each received intravenous injection of either (b) gadolinium-160 (160Gd)-labeled anti-CD68 antibodies or (c) superparamagnetic iron oxide nanoparticles (SPIONs). (d) 160Gd-CD68 antibodies targeted macrophages in the periablational rim, whereas (e) SPIONs were phagocytosed by circulating macrophages moving to the site of inflammation. (f) Magnetic resonance imaging was performed 24 hours after administration of either 160Gd-CD68 or SPIONs, and imaging findings were correlated with histopathology.

Article Snippet: IHC staining for macrophages was performed with a purified rabbit monoclonal anti-CD68 antibody (2449D, MAB101141–100, 1:200; R&D Systems, Minneapolis, Minnesota).

Techniques: Magnetic Resonance Imaging, In Vivo, Injection, Labeling, Imaging, Histopathology

Journal: Journal of vascular and interventional radiology : JVIR

Article Title: MR Imaging–Based In Vivo Macrophage Imaging to Monitor Immune Response after Radiofrequency Ablation of the Liver

doi: 10.1016/j.jvir.2022.11.013

Figure Lengend Snippet: Effects of radiofrequency (RF) ablation–induced immune response at the necrotic margins after hepatic RF ablation in wild-type mice. (a) Photomicrographs of hematoxylin and eosin (H&E)-stained tissue slices (top) showed formation of a time-dependent transitional zone of immune cell infiltration (mean thickness of the rim, 206 μm ± 12). Immunohistochemistry staining with anti-CD68 (bottom) revealed accumulation of macrophages peaking at Day 7 after RF ablation. (b) Representation of percentage cell positivity in a selected 2 × 2-mm2 hotspot area using a positive pixel count algorithm (**P < .01). AB = ablated zone; NL = normal liver; RFA = radiofrequency ablation.

Article Snippet: IHC staining for macrophages was performed with a purified rabbit monoclonal anti-CD68 antibody (2449D, MAB101141–100, 1:200; R&D Systems, Minneapolis, Minnesota).

Techniques: Staining, Immunohistochemistry

Journal: Journal of vascular and interventional radiology : JVIR

Article Title: MR Imaging–Based In Vivo Macrophage Imaging to Monitor Immune Response after Radiofrequency Ablation of the Liver

doi: 10.1016/j.jvir.2022.11.013

Figure Lengend Snippet: In vivo magnetic resonance (MR) imaging–based molecular imaging of immune cell infiltration of macrophages with gadolinium-160 (160Gd)-labeled anti-CD68 antibody. (a) A precontrast T1-weighted MR imaging scan of ablated mouse liver (asterisk) 7 days after radiofrequency ablation. (b) Contrast-enhanced T1-weighted MR imaging of mouse liver 7 days after ablation before systemic delivery of 160Gd-CD68 (baseline). (c) A T1-weighted MR imaging scan (repetition time/echo time, 11,500 ms/5.46 ms) of mouse liver 7 days after ablation and 24 hours after systemic delivery of 160Gd-CD68 showed hypointense periablational rim enhancement (arrows) (d) at the margins of the ablated zone (asterisk). (e, f) Bright-field images of ex vivo imaging mass cytometry of harvested tissue confirmed local deposition of macrophages at the transitional zone (TZ). *ablated liver. The box in Figure 5e indicated the area of magnification for Figure 5f. AB = ablated zone; L = liver; MRI = magnetic resonance imaging.

Article Snippet: IHC staining for macrophages was performed with a purified rabbit monoclonal anti-CD68 antibody (2449D, MAB101141–100, 1:200; R&D Systems, Minneapolis, Minnesota).

Techniques: In Vivo, Imaging, Labeling, Ex Vivo, Mass Cytometry, Magnetic Resonance Imaging

Journal: Cell Death & Disease

Article Title: The ASIC3-M-CSF-M2 macrophage-positive feedback loop modulates fibroblast-to-myofibroblast differentiation in skin fibrosis pathogenesis

doi: 10.1038/s41419-022-04981-9

Figure Lengend Snippet: A Experimental design of fibroblast and macrophage indirect co-culture assay. B Representative images of THP-1 cells and 100 ng/ml PMA-derived M0 macrophages. C Flow cytometric analysis of CD68 in THP-1 cells and 100 ng/ml PMA-derived M0 macrophages. D Immunofluorescence staining of CD206 (green) in experimental and control macrophages after 48 h of co-cultivation. Scale bars, 100 μm. E , F Western blot and RT-qPCR analysis of CD206 protein levels and relative mRNA levels expression in the indicated groups ( n = 3). GAPDH served as a loading control. G Expressions of CD68 and CD206 were determined by flow cytometry. H Quantification of the flow cytometry assay results (n = 3). Data are expressed as the means ± SD. * P < 0.05, ** P < 0.01, and *** P ≤ 0.001. THP-1, human acute monocytic leukemia cell line; PMA, phorbol 12-myrisate 13-acetate; APC, allophycocyanin; FITC, fluorescein-isothiocyanate; GMQ, 2-guanidine-4-methylquinazoline.

Article Snippet: After blocking in goat serum at 37 °C for 30 min, the samples were incubated with antibodies against CD68 (Novus Biologicals, USA, NBP2-32831), CD206 (NovusBiologicals, USA, MAB2534), α-SMA (Novus Biologicals, USA, NBP2-33006), or Collagen I (Novus Biologicals, USA, NB600-408) in Diluent at 4 °C overnight.

Techniques: Co-culture Assay, Derivative Assay, Immunofluorescence, Staining, Control, Western Blot, Quantitative RT-PCR, Expressing, Flow Cytometry

Journal: Cell Death & Disease

Article Title: The ASIC3-M-CSF-M2 macrophage-positive feedback loop modulates fibroblast-to-myofibroblast differentiation in skin fibrosis pathogenesis

doi: 10.1038/s41419-022-04981-9

Figure Lengend Snippet: A Schematic diagram of establishing rabbit ear hypertrophic scar model and experimental plan. B Immunohistochemistry analysis of ASIC3 expression in rabbit ear tissue 21 days after wounding. Scale bars, 100 μm. C Immunofluorescence staining of rabbit ear tissue 21 days after wounding showing the total (CD68+, green) and M2 subtype (CD206+, red) macrophages after treatment by PBS, DMSO, or GMQ. Scale bars, 100 μm. D , E MFI quantification of CD68 and CD206 in (C) ( n = 3). F , G Western blot and RT-qPCR analysis of rabbit ear tissue 21 days after wounding. Total CD206 protein levels and relative mRNA levels expression in control, vehicle, and GMQ groups ( n = 3). H , I Relative mRNA expressions of M1 macrophage markers (iNOS, TNF-α) as evaluated by RT-qPCR ( n = 3). Data are expressed as the means ± SD. * P < 0.05, *** P ≤ 0.001. NS, not significant. iNOS, inducible nitric oxide synthase; TNF-α, tumor necrosis factor alpha.

Article Snippet: After blocking in goat serum at 37 °C for 30 min, the samples were incubated with antibodies against CD68 (Novus Biologicals, USA, NBP2-32831), CD206 (NovusBiologicals, USA, MAB2534), α-SMA (Novus Biologicals, USA, NBP2-33006), or Collagen I (Novus Biologicals, USA, NB600-408) in Diluent at 4 °C overnight.

Techniques: Immunohistochemistry, Expressing, Immunofluorescence, Staining, Western Blot, Quantitative RT-PCR, Control