Journal: Cell reports

Article Title: Multi-modal characterization and simulation of human epileptic circuitry

doi: 10.1016/j.celrep.2022.111873

Figure Lengend Snippet:

Article Snippet: Single-neuron biophysical all-active models , Allen Institute for Brain Science , https://portal.brain-map.org/explore/models/perisomatic-single-neurons.

Techniques: Software, Extraction

Journal: Biosensors

Article Title: The Importance of Multifrequency Impedance Sensing of Endothelial Barrier Formation Using ECIS Technology for the Generation of a Strong and Durable Paracellular Barrier

doi: 10.3390/bios8030064

Figure Lengend Snippet: Schematic illustration demonstrating electrical cell-substrate impedance sensing (ECIS) current flow and modelled parameters. ( A ) An equivalent circuit demonstrating how the electrode, cell layer, and respective media compartments are connected within the ECIS model. Modelling for the paracellular barrier (R b ), basal adhesion (α), and cell membrane capacitance (C m ) integrate different components of the electrode capacitance (C el ), the basal cleft resistance (R cleft ), the cell capacitance and resistance (C c and R c , respectively), and the resistance of apical medium (R m ); ( B ) A schematic of endothelial cells grown on an ECIS electrode. Changes in cell–cell junctions and subcellular adhesion can be measured with ECIS as changes in the current flow at high (>10,000 Hz) and low (4000 Hz) frequencies. A high-frequency current passes through the cell body to couple the capacitive functions of the electrode and cell membrane. Low frequencies take the paracellular route and are resisted by intercellular junctions such as tight junctions and adherens junctions (measured as resistance). Analysis of confluent monolayer properties uses the modelled parameters R b, α, and C m. R b is dependent on the collective sum of the intercellular space and the tightness of cell–cell junctions, α is dependent on the cell radius and subcellular adhesion, and C m models changes in the composition of cell membranes.

Article Snippet: The endothelial barrier resistance was then monitored for at least 48 h, at which point multifrequency data was collected and modelled using ECIS software (Applied Biophysics, Troy, NY, USA).

Techniques: Membrane

Journal: Biosensors

Article Title: The Importance of Multifrequency Impedance Sensing of Endothelial Barrier Formation Using ECIS Technology for the Generation of a Strong and Durable Paracellular Barrier

doi: 10.3390/bios8030064

Figure Lengend Snippet: Immunocytochemistry of brain endothelial junctional space following growth in either Enriched Media or Minimal Media. Cells were seeded at time 0 h at 20,000 cells per 0.3 cm 2 . ( A ) ECIS resistance and R b measurements over 120 h. Cells were grown in either Enriched Media or Minimal Media from T = 0 h, with media changed at T = 48 h (I). Data shown is the mean ± SD (n = 3 wells) of one independent experiment representative of three experimental repeats; ( B ) Cell number count of Hoechst stained nuclei at time point II, obtained through Image J Software analysis. Data shown is the mean ± SEM (n = 18 wells) of 1 independent experiment representative of 3 experimental repeats; ( C ) The junctional space following growth in either Enriched Media or Minimal Media 72 h post seeding. The time point of fixation corresponds to the second vertical dotted line (II) shown on the ECIS traces. Representative GFP/DAPI and GFP monochrome images for the junctional proteins CD144, ZO-1, β-catenin, and α-catenin are shown. Each panel shows cells grown in Minimal Media on the left and cells grown in Enriched Media on the right. The corresponding Alexa Fluor 488 (GFP) monochrome image is shown below each merged image. Green—junctional protein, blue—nuclei. Scale bar is 200 µm. Immunocytochemistry data show one representative image from one independent experiment, which is representative of three experimental repeats. Graphical representations of p values are * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001, **** p ≤ 0.0001.

Article Snippet: The endothelial barrier resistance was then monitored for at least 48 h, at which point multifrequency data was collected and modelled using ECIS software (Applied Biophysics, Troy, NY, USA).

Techniques: Immunocytochemistry, Staining, Software

Journal: Biosensors

Article Title: The Importance of Multifrequency Impedance Sensing of Endothelial Barrier Formation Using ECIS Technology for the Generation of a Strong and Durable Paracellular Barrier

doi: 10.3390/bios8030064

Figure Lengend Snippet: Immunocytochemistry of brain endothelial junctional space following growth in either Enriched Media or Minimal Media. Cells were seeded at time 0 h at 20,000 cells per 0.3 cm 2 . ( A ) ECIS resistance and R b measurements over 120 h. Cells were grown in either Enriched Media or Minimal Media from T = 0 h, with media changed at T = 48 h (I). Data shown is the mean ± S.D. (n = 3 wells) of 1 independent experiment representative of 3 experimental repeats; ( B ) Cell number count of Hoechst stained nuclei at time point II, obtained through Image J Software analysis. Data shown is the mean ± SEM (n = 18 wells) of 1 independent experiment representative of 3 experimental repeats; ( C ) The junctional space following growth in either Enriched Media or Minimal Media 72 h post seeding. The time point of fixation corresponds to the second vertical dotted line (II) shown on the ECIS traces. Representative GFP/DAPI and GFP monochrome images for the junctional proteins CD144, ZO-1, β-catenin and α-catenin are shown. Each panel shows cells grown in Minimal Media on the left and cells grown in Enriched Media on the right. The corresponding Alexa Fluor 488 (GFP) monochrome image is shown below each merged image. Green—junctional protein, blue—nuclei. Scale bar is 200 µm. Immunocytochemistry data shows 1 representative image from 1 independent experiment, which is representative of 3 experimental repeats. Graphical representations of p values are * p ≤ 0.05, ** p ≤ 0.01, *** p ≤ 0.001, **** p ≤ 0.0001.

Article Snippet: The endothelial barrier resistance was then monitored for at least 48 h, at which point multifrequency data was collected and modelled using ECIS software (Applied Biophysics, Troy, NY, USA).

Techniques: Immunocytochemistry, Staining, Software