Journal: Scientific Reports

Article Title: Enhancing cell motility via non-contact capacitively coupled electrostatic field

doi: 10.1038/s41598-024-77384-9

Figure Lengend Snippet: E-field effects on collective migration of T98G monolayers adherent to polystyrene and glass surface tested by scratch assay parallel to the electrodes. Phase contrast micrographs of the same visual field acquired at 200× magnification right after scratching (T = 0 h; left) and after 6 h of EF exposure (T = 6 h; right) at the level of the scratch on the specimen of polystyrene ( A ) and glass ( B ). The yellow area represents the Binary Area Fraction. Scale bars are 250 μm. The black arrow above each micrograph represents the E-field direction from the positive pole to the ground. Mean of the Binary Area Fractions measured from the images scanned along the scratch carried out in six independent experiments (CTL n = 75; EF n = 99) for polystyrene ( C ) and four experiments (CTL n = 36; EF n = 32) for glass ( D ). Blue and red solid lines show the percentage of closure of the scratch along time (20 h) for CTL and EF experiments, respectively. Blue and red transparencies are the relative confidence interval (95%). The inserts on the right side of each graph show the area of data collection relative to the scratch made on monolayers ( C and D ).

Article Snippet: An aliquot, calculated to have a final density of 1 × 10 5 cells/cm 2 , was diluted in 3 ml of supplemented MEM and plated on two different 35 (ø) mm Petri dishes for cell culture: (i) a polystyrene bottom (Falcon ® TC-treated Easy-Grip Style - #353001, Becton Dickinson Labware, USA) vacuum gas plasma treated; (ii) a polystyrene dish with a glass bottom microwell of 10 (ø) mm inserted in the center and gamma irradiated, suitable for fluorescence microscopy (MatTek Corporation, Ashland, MA, USA).

Techniques: Migration, Wound Healing Assay

Journal: Scientific Reports

Article Title: Enhancing cell motility via non-contact capacitively coupled electrostatic field

doi: 10.1038/s41598-024-77384-9

Figure Lengend Snippet: E-field effects on collective migration of T98G monolayers adherent to polystyrene and glass surface tested by scratch assay parallel to the electrodes. Phase contrast micrographs of the same visual field acquired at 200× magnification right after scratching (T = 0 h; left) and after 6 h of EF exposure (T = 6 h; right) at the level of the scratch on the specimen of polystyrene ( A ) and glass ( B ). The two leading edges are highlighted by solid red lines for the ground side and dashed lines for the positive pole side. Scale bars are 250 μm. The black arrow above each micrograph represents the E-field direction from the positive pole to the ground. Mean Distance (µm) performed by the leading edges on the Ground (Edges 1, blue for CTL and red for EF solid lines) and Positive Pole side (Edge 2, blue for CTL and red for EF dashed lines) during the first four hours on the polystyrene (CTL n = 75; EF n = 99) ( C ) and glass (CTL n = 36; EF n = 32) ( D ) surface. Blue and red transparencies are the relative confidence interval (95%). The inserts on the left side of each graph show the area of data collection relative to the scratch made on monolayers ( C and D ). Mean velocity (µm/hours) performed by the leading edges on the Ground (Edges 1, blue for CTL and red for EF solid lines) and Positive Pole side (Edge 2, blue for CTL and red for EF dashed lines) during the first four hours on the polystyrene (CTL n = 75; EF n = 99) ( E ) and glass (CTL n = 36; EF n = 32) ( F ) surface. Blue and red transparencies are the relative confidence interval (95%).

Article Snippet: An aliquot, calculated to have a final density of 1 × 10 5 cells/cm 2 , was diluted in 3 ml of supplemented MEM and plated on two different 35 (ø) mm Petri dishes for cell culture: (i) a polystyrene bottom (Falcon ® TC-treated Easy-Grip Style - #353001, Becton Dickinson Labware, USA) vacuum gas plasma treated; (ii) a polystyrene dish with a glass bottom microwell of 10 (ø) mm inserted in the center and gamma irradiated, suitable for fluorescence microscopy (MatTek Corporation, Ashland, MA, USA).

Techniques: Migration, Wound Healing Assay

Journal: Scientific Reports

Article Title: Enhancing cell motility via non-contact capacitively coupled electrostatic field

doi: 10.1038/s41598-024-77384-9

Figure Lengend Snippet: E-field effects on collective migration of T98G monolayers adherent to polystyrene surface tested by scratch assay perpendicular to the electrodes. Phase contrast micrographs of the same visual field acquired at 200× magnification right after scratching (T = 0 h; above) and after 6 h of EF exposure (T = 6 h; bottom) at the level of the scratch near the ground electrode ( A ), the center ( B ) and the positive pole ( C ). The colored areas represent the Binary Area Fraction. Scale bars are 250 μm. The black arrow above each micrograph represents the E-field direction from the positive pole to the ground. D ) Mean of the Binary Area Fractions calculated from the images scanned near the Ground (black, n = 16), the Center (green, n = 16) and the Positive (red, n = 16) regions along the scratch carried out perpendicular to the electrodes and during 20 h of EF exposure in three independent experiments. Green, black and red transparencies are the relative confidence interval (95%). Blue solid line shows the percentage of closure of the scratch along time for CTL ( n = 75) in the six independent experiments showed in Fig. C, blue transparency is the relative confidence interval (95%). E ) Mean of the Binary Area Fractions measured during EF exposure from the images scanned along the entire scratch carried out perpendicular to the electrodes in the three independent experiments showed in Fig. 5D (red dashed line, n = 48) and parallel to the electrodes in the six independent experiments showed in Fig. C (red solid line n = 99). Red transparencies are the relative confidence interval (95%). The inserts on the right side of each graph show the area of data collection relative to the scratch made on monolayers ( D and E ).

Article Snippet: An aliquot, calculated to have a final density of 1 × 10 5 cells/cm 2 , was diluted in 3 ml of supplemented MEM and plated on two different 35 (ø) mm Petri dishes for cell culture: (i) a polystyrene bottom (Falcon ® TC-treated Easy-Grip Style - #353001, Becton Dickinson Labware, USA) vacuum gas plasma treated; (ii) a polystyrene dish with a glass bottom microwell of 10 (ø) mm inserted in the center and gamma irradiated, suitable for fluorescence microscopy (MatTek Corporation, Ashland, MA, USA).

Techniques: Migration, Wound Healing Assay

Journal: Scientific Reports

Article Title: Enhancing cell motility via non-contact capacitively coupled electrostatic field

doi: 10.1038/s41598-024-77384-9

Figure Lengend Snippet: E-field effects on collective migration of T98G monolayers adherent to polystyrene tested by scratch assay perpendicular to the electrodes. A ) Phase contrast micrographs of the same visual field acquired at 200× magnification right after scratching (T = 0 h; left) and after 6 h of EF exposure (T = 6 h; right) at the level of the scratch near the ground electrode. The two leading edges are highlighted by solid black lines for the upper side and dashed lines for the lower side. Scale bars are 250 μm. The black arrow above each micrograph represents the E-field direction from the positive pole to the ground. B ) Mean of the Distance (µm) performed by the leading edges on the Ground (Edge 1, black solid line and Edge 2, black dashed line) and Positive Pole side (Edge 1, red solid line and Edge 2, red dashed line) during the first two hours. Black and red transparencies are the relative confidence interval (95%). The insert on the left side of the graph shows the area of data collection relative to the scratch in the positive (red dashed ellipse) and ground (black dashed ellipse) sides ( B ). C ) Mean velocity (µm/hours) performed by the leading edges on the Ground (Edges 1, black solid line and Edge 2, black dashed line) and Positive Pole side (Edge 2, red solid line and Edge 2, red dashed line) during the first two hours. Black and red transparencies are the relative confidence interval (95%).

Article Snippet: An aliquot, calculated to have a final density of 1 × 10 5 cells/cm 2 , was diluted in 3 ml of supplemented MEM and plated on two different 35 (ø) mm Petri dishes for cell culture: (i) a polystyrene bottom (Falcon ® TC-treated Easy-Grip Style - #353001, Becton Dickinson Labware, USA) vacuum gas plasma treated; (ii) a polystyrene dish with a glass bottom microwell of 10 (ø) mm inserted in the center and gamma irradiated, suitable for fluorescence microscopy (MatTek Corporation, Ashland, MA, USA).

Techniques: Migration, Wound Healing Assay

Journal: Scientific Reports

Article Title: Enhancing cell motility via non-contact capacitively coupled electrostatic field

doi: 10.1038/s41598-024-77384-9

Figure Lengend Snippet: E-field effects on the movement of single cell adhering to the polystyrene surface tested by tracking along the entire line perpendicular to the electrodes their path for three hours of control and three hours of exposure. A ) and B ) Example of two visual fields acquired in phase contrast at 200× of magnification after 3 h of control conditions (T CTL = 3 h), and after 3 h of E-field exposure (T EF = 6 h). The sample cells and their tracked pathways are visualized in different colors. Scale bars are 200 μm. C ) and D ) Histograms of the Probability Density Function (PDF) for the Path Length (µm) performed by each cell ( n = 280, collected form six independent experiments) monitored during T CTL (Blue) and T EF (Red). The PDF has been fitted by a gamma function (Solid lines). E ) Box Plot of the Path Length (µm) analysis performed by each cell ( n = 280) monitored during T CTL (Blue) and T EF (Red). The non-parametric Kruskal-Wallis statistical test indicated a Chi-square distribution with 1 degree of freedom H = 42.346 that give a corresponding P < 0.00001. F ) Mean Square Displacement (MSD; µm 2 ) performed by the cells population during T CTL (Blue) and T EF (Red) and the relative intervals of confidence (shadow areas).

Article Snippet: An aliquot, calculated to have a final density of 1 × 10 5 cells/cm 2 , was diluted in 3 ml of supplemented MEM and plated on two different 35 (ø) mm Petri dishes for cell culture: (i) a polystyrene bottom (Falcon ® TC-treated Easy-Grip Style - #353001, Becton Dickinson Labware, USA) vacuum gas plasma treated; (ii) a polystyrene dish with a glass bottom microwell of 10 (ø) mm inserted in the center and gamma irradiated, suitable for fluorescence microscopy (MatTek Corporation, Ashland, MA, USA).

Techniques: Control

Journal: Scientific Reports

Article Title: Enhancing cell motility via non-contact capacitively coupled electrostatic field

doi: 10.1038/s41598-024-77384-9

Figure Lengend Snippet: Path tracking of single-cell movement on the polystyrene surface as a function of their relative position with respect to the electrodes. A ) Exposure system geometry and spatial distribution of the E-field divergence on the adherent cell plane (color scale) with T98G considered for the analysis (white square for Ground, n = 107; white triangle for Center, n = 94 and white circles for Positive, n = 79). B ) Histograms of the Probability Density Function (PDF) for the Path Length Variation (%) performed by each cell. C ) Box Plot of the Path Length Variation (%) analysis of the sub-populations performing above the threshold of 50% (Ground n = 35; Center n = 25; Positive Pole n = 27). The non-parametric Kruskal-Wallis statistical test indicated a Chi-square distribution with 2 degrees of freedom H = 2.09 that give a corresponding P = 0.35. D ) Mean Path Speed (µm/h ± SEM) of the sub-populations performing above the threshold of 50% (black line, Ground n = 35; green line, Center n = 24; red line, Positive n = 29) along the time axis (3 h of CTL + 3 h of EF whose starting point is indicated by the grey dashed line) with an image-lapse acquisition frequency of 4/h. P < 0.05 for a two tailed t-Test performed on each time point of Ground ( n = 35) vs. Positive ( n = 29).

Article Snippet: An aliquot, calculated to have a final density of 1 × 10 5 cells/cm 2 , was diluted in 3 ml of supplemented MEM and plated on two different 35 (ø) mm Petri dishes for cell culture: (i) a polystyrene bottom (Falcon ® TC-treated Easy-Grip Style - #353001, Becton Dickinson Labware, USA) vacuum gas plasma treated; (ii) a polystyrene dish with a glass bottom microwell of 10 (ø) mm inserted in the center and gamma irradiated, suitable for fluorescence microscopy (MatTek Corporation, Ashland, MA, USA).

Techniques: Two Tailed Test

Journal: Scientific Reports

Article Title: Enhancing cell motility via non-contact capacitively coupled electrostatic field

doi: 10.1038/s41598-024-77384-9

Figure Lengend Snippet: Polar graph of single-cell movement on the polystyrene surface as a function of their relative position to the electrodes. The track of each T98G cell belonging to the sub-populations with performance above the threshold of 50% (black line, Ground n = 35; green line, Center n = 24; red line, Positive Pole n = 29) during the 3 h of control and the 3 h of E-field exposure are plotted as vectors describing the mean Segment Length (magnitude) and the mean Heading (direction). The yellow dots indicate the averages of X and Y values of each cell reached at the end of the of CTL and EF phases (see Supplementary Information File).

Article Snippet: An aliquot, calculated to have a final density of 1 × 10 5 cells/cm 2 , was diluted in 3 ml of supplemented MEM and plated on two different 35 (ø) mm Petri dishes for cell culture: (i) a polystyrene bottom (Falcon ® TC-treated Easy-Grip Style - #353001, Becton Dickinson Labware, USA) vacuum gas plasma treated; (ii) a polystyrene dish with a glass bottom microwell of 10 (ø) mm inserted in the center and gamma irradiated, suitable for fluorescence microscopy (MatTek Corporation, Ashland, MA, USA).

Techniques: Control