vivo imaging system  (Revvity)

Journal: Bioactive Materials

Article Title: Sustained release PLGA microspheres loaded with a bone-affinity Bmp2 enhance fracture healing and mitigate heterotopic ossification

doi: 10.1016/j.bioactmat.2026.02.050

Figure Lengend Snippet: Design and validation of bone-targeted Bmp2. a. Schematic representation of the Bmp2/D-Bmp2 plasmid constructs and the structure predicted by AlphaFold3. b. Immunofluorescence staining of HEK-293T cells transfected with the plasmids: phalloidin (green), DAPI (blue), and Alexa Fluor 647-conjugated anti-Flag antibodies (red). Scale bar: 10 μm. c. SDS-PAGE of purified proteins. d. Western blot validation of protein expression. e. Bmp2 activity reporter assay: schematic of the luciferase reporter system (left), representative fluorescence images, and statistical analysis of firefly luciferase activity by in vivo imaging system (IVIS) (a.u.: arbitrary units) (right, n = 3 per group). f. qPCR analysis of Bmp2 signaling pathway-related mRNA levels in MC3T3-E1 cells treated with the Bmp2 or D-Bmp2 protein (n = 3 per group). g, h. Western blot (g) and quantification of Bmp2 pathway-related protein expression in treated MC3T3-E1 cells (h) (n = 3 per group). i. Schematic of the HA-coated ELISA plate and HA-binding affinity assay (n = 3 per group). j. Tissue-specific binding assay of Bmp2 or D-Bmp2: Mouse muscle slices (left, scale bar: 100 μm) and undecalcified femur slices (right, scale bar: 200 μm) stained with AF647-conjugated anti-Flag antibodies. The data are presented as the means ± standard deviations (SDs). One-way ANOVA was used for multiple comparisons. Significance levels: ns (not significant), ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001.

Article Snippet: After 24 h, the medium was changed to contain either Bmp2 or D-Bmp2 in DMEM, and the cells were further cultured for another 24 h. After the supernatant was removed, D-luciferin potassium salt (Beyotime, China) was added, and the firefly fluorescence signal was recorded using an in vivo imaging system (IVIS) (PerkinElmer, USA).

Techniques: Biomarker Discovery, Plasmid Preparation, Construct, Immunofluorescence, Staining, Transfection, SDS Page, Purification, Western Blot, Expressing, Activity Assay, Reporter Assay, Luciferase, Fluorescence, In Vivo Imaging, Enzyme-linked Immunosorbent Assay, Binding Assay

Journal: International Journal of Nanomedicine

Article Title: Lactobacillus johnsonii -Derived Extracellular Vesicles Ameliorate Alcohol-Exacerbated Experimental Autoimmune Prostatitis by Inhibiting M1 Macrophage Polarization

doi: 10.2147/IJN.S596237

Figure Lengend Snippet: Alcohol consumption aggravates prostate inflammation and promotes M1 macrophage polarization in EAP mice. ( A ) Schematic of the experimental timeline for EAP induction and alcohol intervention. ( B ) The body weights of mice in the Control, Control+Alcohol, EAP, and EAP+Alcohol groups. ( C and D ) Response frequency to mechanical stimuli and the 50% mechanical withdrawal threshold. ( E ) Prostate inflammation score. ( F ) H&E staining of the anterior and dorsolateral prostate. Scale bars, 100 μm. ( G ) Percentage of CD45-positive staining in prostate immunohistochemistry. ( H ) Immunohistochemical staining of CD45 in prostate tissues. Scale bars, 100 μm. ( I-J ) Relative fluorescence intensity of CD86 and CD206 in prostate tissue. ( K ) Immunofluorescence staining of CD86 and CD206 in prostate tissue. Scale bars, 50 μm. Data are presented as mean ± SD (n = 6 for B-K). *, p < 0.05; ***, p < 0.001.

Article Snippet: The in vivo fluorescence distribution was evaluated 12 hours post-injection using an imaging system from MILabs (Netherlands).

Techniques: Control, Staining, Immunohistochemistry, Immunohistochemical staining, Fluorescence, Immunofluorescence

Journal: International Journal of Nanomedicine

Article Title: Lactobacillus johnsonii -Derived Extracellular Vesicles Ameliorate Alcohol-Exacerbated Experimental Autoimmune Prostatitis by Inhibiting M1 Macrophage Polarization

doi: 10.2147/IJN.S596237

Figure Lengend Snippet: The gut microbiota mediates the effect of alcohol on EAP mice. ( A and B ) Response frequency to mechanical stimuli and the 50% mechanical withdrawal threshold. ( C ) Prostate inflammation score. ( D ) H&E staining of the anterior and dorsolateral prostate. Scale bars, 100 μm. ( E ) Immunohistochemical staining of CD45 in prostate tissues. Scale bars, 100 μm. ( F ) Percentage of CD45-positive staining in prostate immunohistochemistry. ( G ) Immunofluorescence staining of CD86 and CD206 in prostate tissue. Scale bars, 50 μm. ( H ) Relative fluorescence intensity of CD86 and CD206 in prostate tissue. ( I ) Pro-inflammatory cytokine levels in serum. ( J ) Pro-inflammatory cytokine mRNA expression in prostate tissues. Data are presented as mean ± SD (n = 6 for A-I, n = 4 for J). *, p < 0.05; **, p < 0.01; ***, p < 0.001.

Article Snippet: The in vivo fluorescence distribution was evaluated 12 hours post-injection using an imaging system from MILabs (Netherlands).

Techniques: Staining, Immunohistochemical staining, Immunohistochemistry, Immunofluorescence, Fluorescence, Expressing

Journal: International Journal of Nanomedicine

Article Title: Lactobacillus johnsonii -Derived Extracellular Vesicles Ameliorate Alcohol-Exacerbated Experimental Autoimmune Prostatitis by Inhibiting M1 Macrophage Polarization

doi: 10.2147/IJN.S596237

Figure Lengend Snippet: Transplantation of L. john relieves prostate inflammation and pain in EAP mice. ( A and B ) Response frequency to mechanical stimuli and the 50% mechanical withdrawal threshold. ( C ) Prostate inflammation score. ( D ) H&E staining of the anterior and dorsolateral prostate. Scale bars, 100 μm. ( E ) Percentage of CD45-positive staining in prostate immunohistochemistry. ( F ) Immunohistochemical staining of CD45 in prostate tissues. Scale bars, 100 μm. ( G-H ) Relative fluorescence intensity of CD86 and CD206 in prostate tissue. ( I ) Immunofluorescence staining of CD86 and CD206 in prostate tissue. Scale bars, 50 μm. Data are presented as mean ± SD (n = 6 for A-I). *, p < 0.05; **, p < 0.01.

Article Snippet: The in vivo fluorescence distribution was evaluated 12 hours post-injection using an imaging system from MILabs (Netherlands).

Techniques: Transplantation Assay, Staining, Immunohistochemistry, Immunohistochemical staining, Fluorescence, Immunofluorescence

Journal: International Journal of Nanomedicine

Article Title: Lactobacillus johnsonii -Derived Extracellular Vesicles Ameliorate Alcohol-Exacerbated Experimental Autoimmune Prostatitis by Inhibiting M1 Macrophage Polarization

doi: 10.2147/IJN.S596237

Figure Lengend Snippet: LjEVs alleviate prostate inflammation and inhibit M1 macrophage polarization in EAP mice. ( A ) Transmission electron microscopy (TEM) image of LjEVs. Scale bars, 100 nm. ( B and C ) Size distribution and concentration of LjEVs analyzed by nano-flow cytometry (nFCM). ( D and E ) The biodistribution of EVs@ICG versus free ICG was assessed in vivo and ex vivo after administration via tail vein injection. ( F and G ) Response frequency to mechanical stimuli and the 50% mechanical withdrawal threshold. ( H ) Prostate inflammation score. ( I ) H&E staining of the anterior and dorsolateral prostate. Scale bars, 100 μm. ( J ) Immunohistochemical staining of CD45 in prostate tissues. Scale bars, 100 μm. ( K ) Percentage of CD45-positive staining in prostate immunohistochemistry. ( L ) Immunofluorescence staining of CD86 and CD206 in prostate tissue. Scale bars, 50 μm. ( M ) Relative fluorescence intensity of CD86 and CD206 in prostate tissue. ( N ) Pro-inflammatory cytokine levels in serum. ( O ) Pro-inflammatory cytokine mRNA expression in prostate tissues. Data are presented as mean ± SD (n = 6 for F-N, n = 4 for O). *, p < 0.05; **, p < 0.01; ***, p < 0.001.

Article Snippet: The in vivo fluorescence distribution was evaluated 12 hours post-injection using an imaging system from MILabs (Netherlands).

Techniques: Transmission Assay, Electron Microscopy, Concentration Assay, Flow Cytometry, In Vivo, Ex Vivo, Injection, Staining, Immunohistochemical staining, Immunohistochemistry, Immunofluorescence, Fluorescence, Expressing

Journal: Bioactive Materials

Article Title: M2 macrophage-derived exosomes delivering haptoglobin and interleukin-10 plasmids for synergistic therapy of intracerebral hemorrhage

doi: 10.1016/j.bioactmat.2026.01.047

Figure Lengend Snippet: Characterization of M2-exo@HI. (a) Western blot of Tsg101, CD9, and Calnexin expressions in RAW264.7 and M2-exo. (b) Protein bands of RAW264.7, M2-exo, M2-exo@HI, and HI by SDS-PAGE. (c) Fluorescence microscopy images showing Hp-EGFP and IL-10-mCherry expression in 293T cells infected with lipo2000@pBudCE4.1 , lipo2000@HI, and M2-exo@HI respectively, after 24 h. (d,e) Representative TEM images and size distribution profiles of M2-exo and M2-exo@HI. (f) Particle number of M2-exo and M2-exo@HI by NTA measurement. (g) Zeta potentials of M2-exo and M2-exo@HI (n = 3). (h) Drug release profiles of M2-exo@HI at pH 6.5 and pH 7.4, respectively (n = 3). (i) Stability evaluation of M2-exo@HI in PBS at 4 °C by monitoring particle size over time (n = 3). Data are presented as mean ± SD.

Article Snippet: Then mice were anesthetized with 5 % isoflurane and imaged using near-infrared fluorescence in vivo imaging system (IVIS, Caliper Life Sciences, USA) at predetermined time intervals (1 min, 5 min, 10 min, 15min, 2 h, 6 h, 12 h, 24 h).

Techniques: Western Blot, SDS Page, Fluorescence, Microscopy, Expressing, Infection

Journal: Bioactive Materials

Article Title: M2 macrophage-derived exosomes delivering haptoglobin and interleukin-10 plasmids for synergistic therapy of intracerebral hemorrhage

doi: 10.1016/j.bioactmat.2026.01.047

Figure Lengend Snippet: Validation of M2-exo targeting, Hp/IL-10 transfection expression, and Hp/Hb binding. (a) Fluorescence imaging showing cellular uptake of ICG and M2-exo@ICG by M1 microglia. (b,c) Flow cytometry and corresponding quantification of RhB and M2-exo@RhB internalized by M1 microglia (n = 3). (d,e) Schematic illustration and quantitative analysis of the in vitro phagocytosis-release kinetics of M2-exo@RhB in BV2 under ICH-mimicking stimulation (n = 6). (f) Fluorescence images showing Hp and IL-10 expression in M1 microglia treated with M2-exo@HI for 12, 24, 48, 72 h. (g) Mean fluorescence intensity (MFI) quantification of Hp and IL-10 expression (n = 3). (h,i) ELISA measurements of secreted Hp and IL-10 protein levels (n = 3). (j,k) qPCR analysis of relative Hp and IL-10 mRNA expression (n = 3). (l) Western blot detection of Hp and IL-10 protein expression. (m) Densitometric quantification of Hp and IL-10 protein levels from Western blot (n = 3). (n) Co-immunoprecipitation assay confirming the formation of Hp-Hb complex. Data are presented as mean ± SD. Statistical significance was calculated by unpaired Student's t -test (c and e), and one-way ANOVA with Tukey's multiple comparisons test (g-k and m).

Article Snippet: Then mice were anesthetized with 5 % isoflurane and imaged using near-infrared fluorescence in vivo imaging system (IVIS, Caliper Life Sciences, USA) at predetermined time intervals (1 min, 5 min, 10 min, 15min, 2 h, 6 h, 12 h, 24 h).

Techniques: Biomarker Discovery, Transfection, Expressing, Binding Assay, Fluorescence, Imaging, Flow Cytometry, In Vitro, Enzyme-linked Immunosorbent Assay, Western Blot, Co-Immunoprecipitation Assay

Journal: Bioactive Materials

Article Title: M2 macrophage-derived exosomes delivering haptoglobin and interleukin-10 plasmids for synergistic therapy of intracerebral hemorrhage

doi: 10.1016/j.bioactmat.2026.01.047

Figure Lengend Snippet: M2-exo@HI promotes in vitro microglia polarization, BBB repair and neuroprotection. (a) Flow cytometry analysis of M1-type (CD86 + ) and M2-type microglia (CD163 + ) following treatment with different formulations. (b,c) Percentages of CD86 + and CD163 + microglia populations (n = 3). (d–g) The cytokine levels of IL-10, TGF-β, TNF-α, and IL-1β in treated microglia (n = 3). (h) Fluorescence microscopy images showing erythrophagocytosis by microglia across treatment groups. (i) Schematic of the in vitro BBB model assessing FITC-dextran permeability using a transwell assay. (j) Quantitative analysis of FITC-dextran penetration (n = 7). (k) Flow cytometry analysis of neuronal apoptosis across treatments (n = 3). (l) Quantitative analysis of neuronal apoptosis (n = 3). Data are presented as mean ± SD. Statistical significance was tested by one-way ANOVA with Tukey's multiple comparisons test.

Article Snippet: Then mice were anesthetized with 5 % isoflurane and imaged using near-infrared fluorescence in vivo imaging system (IVIS, Caliper Life Sciences, USA) at predetermined time intervals (1 min, 5 min, 10 min, 15min, 2 h, 6 h, 12 h, 24 h).

Techniques: In Vitro, Flow Cytometry, Fluorescence, Microscopy, Permeability, Transwell Assay

Journal: Bioactive Materials

Article Title: M2 macrophage-derived exosomes delivering haptoglobin and interleukin-10 plasmids for synergistic therapy of intracerebral hemorrhage

doi: 10.1016/j.bioactmat.2026.01.047

Figure Lengend Snippet: Targeted delivery and therapeutic gene expression of M2-exo@HI in hemorrhagic brain. (a) In vivo near-infrared fluorescence imaging showing ICG and M2-exo@ICG in mouse brains at various time points post-injection. (b) Average radiation efficiency of ICG in different treatment groups (n = 3). (c) Ex vivo fluorescence imaging of major organs harvested 24 h post-injection. (d) Average radiation efficiency of ICG in different in mouse tissues (n = 3). (e) Time-dependent accumulation of M2-exo@ICG at hematoma regions. (f) Immunofluorescence staining showing co-localization of Hp/IL-10 with astrocytes, microglia, and endothelial cells. (g) Temporal expression profiles of Hp and IL-10 in brain tissues. (h,i) ELISA quantification of Hp and IL-10 protein levels in brain homogenates (n = 3). Data are presented as mean ± SD. Statistical significance was calculated by unpaired Student's t -test (b,d), and one-way ANOVA with Tukey's multiple comparisons test (h,i).

Article Snippet: Then mice were anesthetized with 5 % isoflurane and imaged using near-infrared fluorescence in vivo imaging system (IVIS, Caliper Life Sciences, USA) at predetermined time intervals (1 min, 5 min, 10 min, 15min, 2 h, 6 h, 12 h, 24 h).

Techniques: Gene Expression, In Vivo, Fluorescence, Imaging, Injection, Ex Vivo, Immunofluorescence, Staining, Expressing, Enzyme-linked Immunosorbent Assay