ecis instrument applied biophysics ztheta 96 well array station  (Applied BioPhysics)

Journal: bioRxiv

Article Title: Th17 effector cytokines induce shared and distinct microglial and endothelial cell responses in post-streptococcal encephalitis

doi: 10.64898/2026.02.04.703836

Figure Lengend Snippet: a , Schematic of CD4⁺ T cell differentiation, effector cytokine production, and associated brain pathology in ROR γ t -/- mice following recurrent GAS infections. b, Heatmaps of differentially expressed genes (DEGs) related to BBB function, antigen presentation, and interferon response in olfactory bulb (OB) brain endothelial cells (BECs) from GAS-infected wild-type (WT) and ROR γ t -/- mice. Values are shown as log(z-score); significantly altered genes are indicated in black (adjusted p < 0.05). c, Gene ontology (GO) pathway enrichment analysis of transcripts upregulated and downregulated in ROR γ t -/- versus WT BECs after GAS infections. d, IFNγ concentrations in whole OB lysates from WT PBS (gray), WT GAS (blue), and ROR γ t -/- GAS (green) mice after two or five infections. Statistical comparisons were performed using one-way ANOVA (ns, p > 0.05; p < 0.05). e, Heat maps of DEGs related to antigen presentation, homeostatic/DAM programs, and cytokine/chemokine signaling in OB microglia from GAS-infected WT and ROR γ t -/- mice (log(z-score); adjusted p < 0.05 shown in black). f, GO pathway enrichment analysis of transcriptional changes in ROR γ t -/- versus WT microglia following GAS infections. g, Representative flow cytometry plots of CD74 and MHC class II (I-A/I-E) expression in WT and ROR γ t -/- microglia after GAS infections (n = 3,640 and 4,657 cells, respectively). h, Quantification of microglial surface expression of antigen presentation markers in WT PBS, WT GAS, and ROR γ t -/- GAS mice. Statistical comparisons were performed using one-way ANOVA (ns, p > 0.05; p < 0.05; * p < 0.01). i, j, Transendothelial electrical resistance (TEER) measurements in primary mouse BEC monolayers following cytokine treatment, shown as representative ECIS traces ( i ) and area-under-the-curve (AUC) quantification ( j ). Gray shading indicates treatment period. Data are mean ± SEM (n = 5 replicates from 3 independent experiments). Mixed-effects analysis ( p < 0.05; * p < 0.01). k, l, Relative transwell permeability of primary mouse BEC monolayers to albumin - AF594 following cytokine treatment. Untreated cells were used as reference, and LPS served as a positive control. Data are mean ± SEM (n = 3 independent experiments). Repeated-measures one-way ANOVA ( p < 0.05). m, n, TEER measurements in primary human brain microvascular endothelial cells (HBMECs), shown as representative ECIS traces ( m ) and AUC quantification ( n ). Data are mean ± SEM (n = 6 replicates from 3 independent experiments). Mixed-effects analysis (*** p < 0.0001).

Article Snippet: Transendothelial electrical resistance (TEER) was measured in real time using an electric cell-substrate impedance sensing (ECIS) instrument (Applied BioPhysics, ZTheta 96 Well Array Station) as previously described [ ]. mBECs and HBMECs were plated on 96-well plates containing electrode arrays (Applied BioPhysics, 96W20idf).

Techniques: Cell Differentiation, Immunopeptidomics, Olfactory, Infection, Flow Cytometry, Expressing, Permeability, Positive Control

Journal: Critical Care

Article Title: Effect of therapeutic plasma exchange on acquired hypocholesterolemia in patients with septic shock: a post hoc analysis of the two exchange trials

doi: 10.1186/s13054-025-05790-0

Figure Lengend Snippet: Impact of TPE on acquired hypocholesterolemia in septic shock patients and effect of a cholesterol concentrate on endothelial permeability in vitro. Panel A shows serum concentrations of ( a ) total cholesterol (TC), ( b ) LDL-cholesterol, ( c ) HDL-cholesterol and ( d ) triglycerides as violin plots at study inclusion and 6 h after randomization in patients with septic shock who received either standard of care (SOC) alone or SOC in combination with therapeutic plasma exchange (TPE). Blood samples were drawn at study inclusion/randomization and after 6 h. For blood sampling serum was obtained by centrifugation, divided into aliquots and stored at − 80° until assayed. Circulating TC, HDL-, LDL-C, and triglyceride blood concentrations were quantified using enzymatic colorimetric tests on a cobas 8000 automated platform (Roche Diagnostics, Mannheim, Germany). Panel B demonstrates the effect of cholesterol concentrate on endothelial permeability in vitro. Human umbilical vein endothelial cells (HUVECs) were grown in either low serum media (control medium, 0.625%) or medium containing cholesterol concentrate (cholesterol medium, 0.5 nM) ( a ). Additionally, cells were either stimulated with 50 ng/ml TNFa ( b ) or 5 nM Thrombin ( c ) and transendothelial electrical resistance (TER) was measured at different time points by an electric cell-substrate impedance sensing (ECIS) system. Values were plotted over time. Resistance data were normalized to the resistance values at time of media change.Impedance measuring was performed with the ECIS ® Z-Theta instrument (Applied Biophysics Inc, NY, USA) to monitor the effects of total cholesterol on the barrier integrity of HUVECs over time. Paired-t-test or Wilcoxon signed-rank test was used as appropriate for within-group effects between the chosen two time points (randomization, 6 h after randomization). Comparisons between groups were analyzed by means of Mann-Whitney U test. Wilcoxon matched-paired signed rank tests were performed for comparing cholesterol and control longitudinal TER results in the ECIS ex-vivo experiments. For all statistical analyses a two-tailed p-value < 0.05 was considered statistically significant. GraphPad Prism 7 (Graph Pad, La Jolla, CA, USA), SPSS Statistics Version 25 (SPSS Inc., Chicago, IL, USA) and the R environment for statistical computing version 4.1.2 (R Foundation for Statistical Computing, Vienna, Austria) were used for data analysis and graph generation

Article Snippet: Resistance data were normalized to the resistance values at time of media change.Impedance measuring was performed with the ECIS ® Z-Theta instrument (Applied Biophysics Inc, NY, USA) to monitor the effects of total cholesterol on the barrier integrity of HUVECs over time.

Techniques: Permeability, In Vitro, Clinical Proteomics, Sampling, Centrifugation, Control, Electric Cell-substrate Impedance Sensing, MANN-WHITNEY, Ex Vivo, Two Tailed Test