content imaging system  (Revvity)

Journal: Bioactive Materials

Article Title: Precisely regulated physically-crosslinked carriers enable synergetic release of bioactive factors for MSC-mediated cartilage regeneration

doi: 10.1016/j.bioactmat.2026.01.009

Figure Lengend Snippet: Characterization of microspheres. (a) Scanning electronic microscopic (SEM) observations shows the morphology and surface structure of SF microspheres with recrystallization (assembly) time (t) of 0 h, 24 h and 48 h. (b) Fourier transform infrared spectroscopy (FTIR) analysis showing the different second structure of SF (β-sheet, random coil/helix and β-turn) content in different recrystallization time. (c) Matched curve of β-sheet fraction with assembly time. Illustrations showed the morphology of SF microspheres with recrystallization (assembly) time (t) of 0 h, 24 h and 48 h, as well as the Fourier transform infrared spectroscopy (FTIR) analysis showing the change of β-sheet content in different recrystallization time. (d, e) Cumulative release curve of MAP and KGN from PSF and KSF with different β-sheet fraction respectively. (f) The contact angle with water of SF with different β-sheet fraction showing the wetting property. (g) The degradation of SF with different β-sheet fraction. (h) Comparison of release kinetics for drug releasing strategies between our approach (red line) and previous studies reported . The illustration showed the comparison of release kinetics for MSC recruiting strategies between our approach (gray area) and previous studies (purple points). (i) In vivo retention of PSF microspheres injected in the joint cavity visualized by in vivo imaging system (IVIS). (j) Fluorescent intensity (Taking the base-10 logarithm) at different timepoint after injection. ∗ p < 0.05; ∗∗ p < 0.01; ∗∗∗ p < 0.001; ∗∗∗∗ p < 0.0001.

Article Snippet: Immunofluorescent staining was performed with Aggrecan primary antibody (13880-1-AP, Proteintech, USA), ActinGreen ( R37110 , Thermo, USA) and DAPI (Solarbio, China) and observed with 3D reconstruction under high content imaging system (PerkinElmer, Operetta CLS, USA).

Techniques: Recrystallization, Fourier Transform Infrared Spectroscopy, Spectroscopy, Comparison, In Vivo, Injection, In Vivo Imaging

Journal: Acta Neuropathologica Communications

Article Title: Tumor-like proliferation of CCM3 knockout endothelial cells: insights from semaxinib treatment and transcriptome profiling of co-cultures

doi: 10.1186/s40478-026-02283-1

Figure Lengend Snippet: Screening of a cytokine inhibitor library in co-cultures of CCM3 KO and WT iECs. A Scheme of the library screening approach (Created in BioRender. Pilz, R. (2026) https://BioRender.com/lhhhtxj ). B Scheme of the image and data analysis strategy (Created in BioRender. Pilz, R. (2026) https://BioRender.com/o4wswi1 ). The Operetta CLS High-Content Imaging System was used to calculate the total number of nuclei, to allocate the cells to red or green fluorescence (colored outlines of the cells), and to quantify the percentage of green cells of the total cell count per well. The total cell count was used to exclude cytotoxic compounds. Then, statistical analysis was performed to identify hits. C Example of the readout strategy. The first two panels show fluorescence imaging of mEGFP (labeled cytoplasm of AICS-0036 cells), mTagRFP-T (labeled plasma membrane of AICS-0054 cells), and Hoechst 33342 (nuclei) (scale bar = 500 μm). The two panels on the right illustrate the software’s detection and allocation strategy, with an enlarged section for clarity. Nuclei of AICS-0036 cells are outlined in green, while nuclei of AICS-0054 cells are outlined in red. D Representative illustrations of the high-content imaging of plates from one experimental run. The library (compounds) was tested in co-cultures of CCM3 KO (mEGFP) and WT (mTagRFP-T) iECs. DMSO-treated AICS-0036 CCM3 KO/AICS-0054 WT and AICS-0036 WT/AICS-0054 WT co-cultures were used as controls (two columns on the right). E Results of the compound screening showing the percentage of green KO cells from the total cell count for each compound (bars) after excluding cytotoxic compounds. Data are presented as means and SD ( n = 4). All conditions that showed a proportion of green KO cells higher or lower than 50% (purple lines) of the DMSO-treated KO/WT control (highlighted in red) are highlighted in blue. Statistical analysis was performed using one-way ANOVA with Dunnett’s correction (* = Padj < 0.05; *** = Padj < 0.001). All compound-treated conditions were compared against the DMSO-treated control

Article Snippet: Quantification of cell counts was performed using the Operetta CLS High-Content Imaging System (PerkinElmer) or FIJI v.1.54 (ImageJ).

Techniques: Library Screening, Imaging, Fluorescence, Cell Characterization, Labeling, Clinical Proteomics, Membrane, Control

Journal: Acta Neuropathologica Communications

Article Title: Tumor-like proliferation of CCM3 knockout endothelial cells: insights from semaxinib treatment and transcriptome profiling of co-cultures

doi: 10.1186/s40478-026-02283-1

Figure Lengend Snippet: Semaxinib’s effect on CCM3 KO and WT iEC proliferation in 2D co-culture and vascular organoids. A , B Co-cultures of CCM3 KO (mEGFP) and WT (mTagRFP-T) iECs were treated with semaxinib (A) or (Z)-semaxinib (B) at various concentrations and analyzed after six days to assess the dose-dependent effects on proliferation. DMSO-treated co-cultures were used as controls. Representative fluorescence images (scale bar = 500 μm) and the proportion of CCM3 KO cells normalized to the DMSO control and presented as means and SD ( n = 3) are shown for both inhibitors. One-way ANOVA followed by Dunnett’s multiple comparisons test was used for statistical analysis (** = Padj < 0.01; *** = Padj < 0.001). C Co-cultures of CCM3 KO and WT iECs were treated with 10 µM semaxinib or DMSO (control) and fixed on days 0, 1, 2, 3, 4, 5, and 6. Representative fluorescence images are shown (scale bar = 500 μm). The number of WT and KO cells was quantified using the Operetta CLS High-Content Imaging System and is displayed as means and SD ( n = 6). Two-way ANOVA with Tukey’s post hoc test for multiple comparisons was used (** = Padj < 0.01; *** = Padj < 0.001). D Representative images of mosaic vascular organoids generated from co-cultures of CCM3 KO and WT iPSCs at a ratio of 1:19 are shown after treatment with DMSO or semaxinib (scale bar = 200 μm). Mean mEGFP intensity normalized to the DMSO control is displayed as individual data points and means ( n = 75–88). Statistical significance was assessed using a two-tailed unpaired t-test (*** = P < 0.001)

Article Snippet: Quantification of cell counts was performed using the Operetta CLS High-Content Imaging System (PerkinElmer) or FIJI v.1.54 (ImageJ).

Techniques: Co-Culture Assay, Fluorescence, Control, Imaging, Generated, Two Tailed Test