Claims 27
From the paper Sub-microsecond optical measurements of cell membrane charging and lesioning by pulsed electric fields · claims file · checked 2026-10-21: ready
- fig7a-test-pulse-cosine-up-to-1000
After a conditioning pulse of up to 1000 V/cm, polarization by the 150 V/cm test pulse 5 us later still follows a cosine of the angle to the field, while 1200 and 1400 V/cm conditioning pulses markedly reduce charging at the cell poles.
Polarization still followed a cosine function after conditioning PEFs of up to 1000 V/cm. However, 1200 and 1400 V/cm conditioning PEFs markedly hindered charging by the test pulse at cell poles, indicating a persistent membrane permeabilization.
Results, 3.7, paragraph 3; Figure 7AHow it will be judged
Per conditioning strength, the test-pulse dF profile over the 360 perimeter points, averaged over the n = 5 cells (Methods 2.3; Fig. 7A legend), is fit by least squares with a cosine of the angle to the field (A cos(theta - phi)), as Fig. 7A fits it; the benchmark is the fit after the 150 V/cm conditioning pulse (Fig. 7A legend). "Follows a cosine" has no stated measure. It is not read as matching the benchmark amplitude, since the paper puts the 1000 V/cm test response at ~80% of control and still calls it a cosine. Readings - (a) the mean pole dF (+-5 degree arcs) at each pole lies within the s.e. outline of the profile (Fig. 7A legend, gray outline) around the strength's own cosine fit at that pole; (b) pole dF does not differ from the benchmark by the two-sided t-test of Fig. 7B at p < 0.05 (readings of that test as in fig7b-test-pulse-unchanged-to-760). "Markedly hindered" at 1200 and 1400 V/cm is pole dF below the benchmark at p < 0.05 by that test (as in fig7b-test-pulse-reduced-1200-1400). Reproduced when, under a reading, every conditioning strength up to 1000 V/cm follows a cosine and 1200 and 1400 V/cm are hindered at the poles. The cosine amplitude and R^2 of every strength are reported.
- fig7b-conditioning-pulse-below-linear
During the electroporating pulse, pole polarization falls below the linear scaling expected for an undamaged membrane, 1.5-fold below at 560 V/cm and 2-fold at 760 V/cm and further at stronger fields, significantly for every strength of 560 V/cm or more.
Actual measurements fell 1.5 times below this line for 560 V/cm PEFs ... and diverged further for more intense PEFs (p < 0.01 for all datapoints ≥560 V/cm, one sample t-test)
Results, 3.7, paragraph 5; Figure 7B, top panelHow it will be judged
The paper takes these data from the Fig. 4B experiments (Di-8-ANEPPS, single 1-us pulses, imaged at 900 ns, n = 5), so they may be in another dandiset. Pole dF in +-5 degree arcs, from collapsed baseline and polarization images (Methods 2.3); dF at 150 V/cm taken as 100%, linear expectation E/150 x 100% (Fig. 7B legend). Open, with readings. Which pole the 1.5 and 2-fold refer to - (a) cathodic, (b) anodic, (c) the mean of both. How the 150 V/cm normalization is applied - (i) per cell, each cell's values divided by its own 150 V/cm value, and the one-sample t-test of the per-cell values against E/150 x 100%; (ii) on group means, with the one-sample t-test of per-cell dF x 150/E against the group mean at 150 V/cm. Two-sided (Fig. 7B legend, "two-way"). Reproduced when, under one pole reading and one normalization reading, both ratios are within tolerance and every strength of 560 V/cm or more differs from the line at p < 0.01.
- fig7b-test-pulse-unchanged-to-760
Polarization by the test pulse 6 us after the conditioning pulse does not differ from the undamaged control for conditioning pulses up to 760 V/cm, and is at about 80% of control after 1000 V/cm, a difference that is not significant.
measurements 6 μs later (test pulse) did not differ from the control up to 760 V/cm strength of the conditioning PEF; they were at ~80% of the control reading after 1000 V/cm conditioning PEF but the difference was not statistically significant
Results, 3.7, paragraph 5; Figure 7B, bottom panelHow it will be judged
A claim of absence, judged on the paper's test, a two-sided t-test at p < 0.05 for the difference from the undamaged membrane, n = 5 (Results 3.7; Fig. 7B legend). Pole dF in +-5 degree arcs at 900 ns into the test pulse; the test pulse after a 150 V/cm conditioning pulse is the control, taken as 100% (group mean; the legend draws the control as a gray band between dashed lines, so it keeps its spread). The legend does not say whether the two-sided t-test pairs cells, which each received every conditioning strength; readings (i) paired t-test of per-cell dF against the same cell's control dF; (ii) unpaired two-sample t-test. A one-sample t-test of per-cell normalized values against 100% does not fit the legend's control band and is shown as a variant only. Each pole is tested. Reproduced when no conditioning strength up to 760 V/cm differs from control, and 1000 V/cm does not differ either; not reproduced when any does. When readings (i) and (ii) give different outcomes the claim is methods_underspecified, with both shown. The 80% value is checked under the pole readings (a) cathodic, (b) anodic, (c) mean of both. Effect sizes are reported.
- fig7b-test-pulse-reduced-1200-1400
Conditioning pulses of 1200 and 1400 V/cm reduce charging by the test pulse 6 us later to about 70% and 50% of control (p < 0.05 and p < 0.01).
Still more intense PEFs of 1200 and 1400 V/cm reduced charging by the test pulse to ~70% and ~ 50%, respectively (p < 0.05 and p < 0.01, two-sided t-test)
Results, 3.7, paragraph 5; Figure 7B, bottom panelHow it will be judged
Measures and test readings as in fig7b-test-pulse-unchanged-to-760 (test (i) paired or (ii) unpaired; pole (a) cathodic, (b) anodic, (c) mean of both). Reproduced when, under one combination, both values are within tolerance and the 1200 and 1400 V/cm differences reach p < 0.05 and p < 0.01; partially reproduced when both are reduced and significant but a value is outside tolerance.
- fig7c-conductivity-drops-10-fold
Effective membrane conductivity at the poles falls by about 10-fold (range 7 to 20) within 6 us after the electroporating pulse, for most datapoints.
Fig. 7C shows a profound reduction in the λm,eff within just 6 μs after the electroporating pulse. For most datapoints, it dropped by a factor of 10 (range: 7–20).
Results, 3.7, paragraphs 6 and 7; Figure 7CHow it will be judged
As the paper does it (Results 3.7, paragraph 6; Fig. 7C legend), lm,eff is interpolated from the group mean values of Fig. 7B, per pole and conditioning strength, with Eq. (2) and the stated parameters. During the conditioning pulse the ratio of measured to calculated IMP is the mean pole dF over the linear expectation E/150 x 100%; at the test pulse it is the mean pole dF over the control (100%). A negative dF is omitted and a value above 100% is set to 100%, as the paper did. The fold drop is conditioning lm,eff over test lm,eff, for strengths of 560 V/cm and above. "Most datapoints" is read as more than half of the pole and strength pairs for which the test lm,eff is above the intact value (a ratio below 100%; a pair with the test at 100% has no finite fold). Reproduced when the median of those folds is within 9 to 11 and more than half lie within 6.3 to 22 (the range widened by 10% of each bound); partially reproduced when the median is within 6.3 to 22 but not 9 to 11. Every fold is reported.
- fig7c-full-reseal-mild-residual-strong
After mildly electroporating pulses (570 and 760 V/cm) the membrane reseals fully within 6 us, while after stronger pulses increased conductivity persists for at least 6 us; the cathodic pole is more conductive than the anodic during the pulse, but the test pulse finds no difference between the poles.
While the cathodic pole became more conductive than the anodic pole during the electroporating pulse (Fig. 7B, see also Figs. 1 and 4B–D), no difference between the poles was detected by the test pulse. For mildly electroporating PEFs of 570 and 760 V/cm, membrane resealed fully (within the accuracy of measurements) within just 6 μs. With more intense PEFs, the residual increased conductivity persisted for at least 6 μs
Results, 3.7, paragraph 7; Figure 7CHow it will be judged
lm,eff as in fig7c-conductivity-drops-10-fold. Full reseal at 570 and 760 V/cm is judged as the paper judges it, by the test-pulse pole dF not differing from control in the two-sided t-test (fig7b-test-pulse-unchanged-to-760). Residual conductivity after the stronger pulses (1000, 1200 and 1400 V/cm; the abstract says residual conductivity persisted after PEFs above 780 V/cm) is judged as the paper judges it in Fig. 7C, by the test-pulse lm,eff lying above the intact value (test dF below 100% of control) at each of them; the significance of fig7b-test-pulse-reduced-1200-1400 is reported, and the paper calls the 1000 V/cm reduction not significant. Pole difference during the pulse - the cathodic lm,eff exceeds the anodic at most conditioning strengths of 1000 V/cm and above (Results 3.4, paragraph 4, gives both poles about equally permeabilized at 570 and 780 V/cm and the cathodic reduction larger above them; the cathodic 1400 V/cm point counts as higher if its dF is negative, which the paper reads as no charging). No pole difference at the test pulse - a two-sided paired t-test of cathodic against anodic normalized test dF, per strength, finds none at p < 0.05 (the paper names no test for this; the paired t-test is the Methods' Student's t-test applied to the two poles of the same cells; the unpaired t-test is shown as a variant). Reproduced when all four parts hold; partially reproduced when the reseal and residual parts hold.
- fig1-poles-leaky-equator-intact
With 1-us pulses, polarization grows with field strength up to 570 to 780 V/cm; at stronger fields the poles become leaky and show reduced or absent polarization, while regions closer to the equator remain polarized and may exceed the poles.
Still stronger electric fields produce a distinct pattern of reduced or absent polarization at the poles (where the membrane becomes too leaky to accumulate the charge) while regions closer to the equator remain relatively intact and may polarize more than the poles
Figure 1 legend; Results, 3.1, paragraph 1How it will be judged
Fig. 1 shows images of Di-8-ANEPPS cells at 900 ns into 1-us pulses of 43 to 1400 V/cm; the same protocol gives the data of Fig. 4B-C, on which this is judged. Pole dF in +-5 degree arcs from collapsed images, group means. Reproduced when cathodic pole dF is largest at 570 or 780 V/cm and lower at each of 1000 to 1400 V/cm, and at 1400 V/cm |dF| off the pole exceeds |dF| at the cathodic pole. "Closer to the equator" fixes no angle; readings, arcs of +-5 degrees centred (a) 30, (b) 45, (c) 60 degrees from the cathodic pole; the claim holds under any of them.
- fig2a-fluovolt-saturation
FluoVolt dF rises in proportion to field strength until apparent saturation at 66% (255 mV) at 360 V/cm at the cathodic pole and -83% (-320 mV) at 570 V/cm at the anodic pole; stronger fields do not raise it further and lower it at the cathodic pole.
ΔF increased proportionally to the electric field strength until reaching apparent saturation at 66% ΔF (255 mV IMP) at 360 V/cm at the cathodic pole, and −83% ΔF (−320 mV IMP) at 570 V/cm at the anodic pole
Results, 3.2, paragraph 2; Figure 2AHow it will be judged
Fig. 2 method (Methods 2.3) - per image, mean F in +-5 degree pole arcs, F0 the mean of the 40 baseline images of the set, dF averaged over the 60 pulse images and over the n = 4 cells. Reproduced when the cathodic maximum over strengths is at 360 V/cm and the anodic minimum at 570 V/cm, both values within tolerance, and the cathodic dF at 1000 V/cm and above is below its 360 V/cm value. Saturation at a neighbouring strength with both values in tolerance is partially reproduced.
- fig2a-fluovolt-stable-within-set
FluoVolt baseline fluorescence is stable without pulses (no significant bleaching), and the response stays stable through the 60 pulses of each set even at strongly electroporating fields of 1000 V/cm and above.
Field strengths of 1000 V/cm and higher caused strong electroporation (see Section 3.5), but the dye response in each set stayed stable through the 60 pulses applied
Results, 3.2, paragraph 2; Figure 2AHow it will be judged
Per cell and set, pole dF per image as in fig2a-fluovolt-saturation. The paper gives no measure of stability. Readings, each tested on the group means at 1000, 1200 and 1400 V/cm - (a) the mean of the first 10 pulse images against the last 10 (10 is the window of Fig. 4B dotted lines), two-sided paired t-test over cells at p < 0.05; (b) the linear regression slope of dF on image index over the 60 images (Methods 2.5, Grapher regression), tested against zero at p < 0.05. Reproduced when neither finds a change at either pole in any of these sets (a claim of absence; both readings are shown and the effect sizes reported), and the baseline images likewise show no trend.
- fig2b-di8-climbs-then-declines-in-set
Di-8-ANEPPS dF keeps rising up to 570 V/cm at both poles, and responses at 780 V/cm and above decrease gradually within each set of 60 pulses.
ΔF kept climbing up to the electric field strength of 570 V/cm at both cell poles ... responses to PEFs at and above 780 V/cm decreased gradually in each set of 60 images
Results, 3.2, paragraph 3; Figure 2BHow it will be judged
Fig. 2 method as in fig2a-fluovolt-saturation, Di-8-ANEPPS, n = 5. Part 1 - the group mean |dF| at each pole increases from each strength to the next up to 570 V/cm. Part 2 - "decreased gradually" is judged under the readings of fig2a-fluovolt-stable-within-set ((a) first 10 against last 10 pulse images, (b) regression slope), at 780 V/cm and each stronger strength, by the group mean |dF| being lower at the end of the set (a negative slope). The paper reports no test for this decline and calls the like decline of Fig. 2C at 1000 V/cm (n = 8) not significant, so significance is reported, not required. The text names no pole; readings (i) both poles, (ii) the cathodic pole (Discussion, point (a), cites Fig. 2B for the cathodic pole). Reproduced when part 1 holds and part 2 holds under one window and one pole reading at most of those strengths.
- fig2cd-stable-at-150-230
Repeated alternating sets of 150 and 230 V/cm pulses give dF responses that stay constant within and between sets, up to 800 flashes and 480 pulses before the 1000 V/cm exposure.
Repeated sets of PEFs at 150 and 230 V/cm, delivered in alternation, produced ΔF responses that stayed constant both within each set and between the sets
Results, 3.2, paragraph 4; Figure 2C and 2DHow it will be judged
A claim of absence, judged on the paper's test, a two-tailed paired t-test at p < 0.05 (Fig. 2D legend) of each set's dF averaged over its 60 pulse images against the first set at the same field strength, n = 8 cells, per pole. Reproduced when no set before the 1000 V/cm exposure differs from the first; not reproduced when any does. Within-set stability is shown under the readings of fig2a-fluovolt-stable-within-set.
- fig2c-within-set-decline-1000
At 1000 V/cm, dF at the cathodic pole declines from 40 +- 10% at the start of the set to 26.6 +- 8% at its end, a trend that is not significant.
at an electroporating field strength of 1000 V/cm, ΔF at the cathodic pole decreased from 40 ± 10% at the beginning of the set to 26.6 ± 8% at the end of it. Although not statistically significant (p > 0.05)
Results, 3.2, paragraph 4; Figure 2CHow it will be judged
Tolerances are the reported s.e. Fig. 2 method, n = 8. "Beginning" and "end" of the set are not defined; readings (a) the first and the last pulse image; (b) the means of the first 10 and last 10 pulse images. The test is not named; the paired two-tailed t-test of Fig. 2D is used. Reproduced when, under one reading, both values are within tolerance, the end value is lower, and the test gives p > 0.05.
- fig2d-lasting-drop-after-1000
dF averaged over each set stays nearly constant before electroporation but drops by about 20% in the sets acquired after the 1000 V/cm exposure (p < 0.05).
ΔF values averaged across each set of 60 PEFs stayed nearly constant before the electroporation, but dropped by about 20% in the sets acquired after it (Fig. 2D, p < 0.05)
Results, 3.2, paragraph 4; Figure 2DHow it will be judged
Per cell and set, dF averaged over the 60 pulse images, per pole; each 150 and 230 V/cm set after the 1000 V/cm exposure compared with the first set at the same strength (dashed lines of Fig. 2D) by a two-tailed paired t-test, n = 8. Which pole the 20% refers to is not said; readings (a) cathodic, (b) anodic, (c) both. The size has two readings from the paper - the text's "about 20%" (20 +- 2) and the Fig. 2D legend's "small (<=20%)" (a drop above zero and at most 22%, the bound widened by 10%). Reproduced when, under one pole reading and one size reading, the mean relative drop over the post-exposure sets is within it and at least one post-exposure set differs at p < 0.05.
- fig3-dye-calibration-slopes
Fit through the origin over -150 to +150 mV of calculated IMP, FluoVolt dF changes by 26% and Di-8-ANEPPS by 9.1% per 100 mV, with correlation coefficients 0.96 and 0.78.
The correlation coefficients of the fits equaled 0.96 for FluoVolt and 0.78 for Di-8 ANEPPS (due to a lower signal-to-noise ratio), with respective slope coefficients of 26% and 9.1% ΔF per 100 mV of IMP
Results, 3.3; Figure 3How it will be judged
As Results 3.3 and the Fig. 3 legend fix it - every individual dF at every perimeter point of every cell at 43 to 360 V/cm (1-us pulses, 900 ns), against IMPC for that point (Eq. 1, the cell's R and tau from the fit of [20]); points with IMPC within -150 to +150 mV fit by least squares with a line through the origin. The correlation coefficient is not defined for a fit through the origin; readings (a) Pearson r of dF and IMPC over those points; (b) the square root of the uncentred R^2 of the fit through the origin. Reproduced when both slopes are within tolerance and, under one reading, both correlation coefficients are. The outside input is the tau-radius fit of Semenov et al. 2025, Bioelectrochemistry 168, 109143 (ref. [20]); at 900 ns into the pulse its effect is small, and the result with tau set to 0 is shown as a variant.
- fig3-fluovolt-nonlinear-di8-linear
FluoVolt departs from linearity already at +-200 mV of calculated IMP, while Di-8-ANEPPS stays essentially linear from -500 to +500 mV, so the early FluoVolt departure reflects the dye's range and not electroporation.
The actual measurements with FluoVolt gradually became non-linear already at ±200 mV, while those for Di-8-ANEPPS stayed essentially linear throughout the tested range from −500 to +500 mV
Results, 3.3; Figure 3How it will be judged
Data and fit as in fig3-dye-calibration-slopes; the paper smooths all points with LOESS and compares it with the line. The LOESS span is not given; readings, spans of (a) 0.3 and (b) 0.5 of the data. Linearity has no stated measure; the measure used is the LOESS value as a fraction of the linear-fit value. Reproduced when, under one span, the FluoVolt fraction falls below 0.9 at |IMPC| of 200 to 300 mV at both signs, and the Di-8-ANEPPS fraction stays within 0.9 to 1.1 from -500 to +500 mV (wherever data exist). The 0.9 to 1.1 band is the protocol's default 10% tolerance, not the paper's.
- fig4a-fluovolt-limit
With FluoVolt, measured and calculated IMP match only at 43 and 150 V/cm; stronger pulses give a maximum dF of 71 +- 10% at the cathodic and -83.5 +- 5% at the anodic pole, and cathodic polarization falls from 71% at 360 V/cm to 45 to 50% at 1000 to 1400 V/cm.
this assumption held only for the applied fields of 43 and 150 V/cm, where measured and calculated IMP matched. With stronger PEFs, maximum ΔF reached 71 ± 10% and −83.5 ± 5% at the cathodic and anodic poles, respectively ... polarization at the cathodic pole decreased from 71% at 360 V/cm to 45–50% at 1000–1400 V/cm
Results, 3.4, paragraph 1; Figure 4AHow it will be judged
FluoVolt, 1-us pulses, n = 4; pole dF in +-5 degree arcs from collapsed images, means over cells (Methods 2.3). Tolerances for the maxima are the reported s.e. The maxima are the extreme group means over strengths. The 45 to 50% range holds when each of the 1000, 1200 and 1400 V/cm cathodic means lies in 40.5 to 55. "Matched" at 43 and 150 V/cm is judged by the pole IMP (dF / 26 x 100 mV) within 10% of the mean IMPC (protocol default), and at 230 V/cm and above by it falling outside. Reproduced when all hold.
- fig4b-di8-threshold-570
With Di-8-ANEPPS, measured IMP falls below the calculated value when the field reaches 570 V/cm, as dF reaches its maximum of 57 +- 14% at the cathodic and -55 +- 4% at the anodic pole, over 600 mV, twice the FluoVolt value.
The measured IMP decreased below the calculated value when the electric field reached 570 V/cm, concurrently with reaching the maximum ΔF of 57 ± 14% at the cathodic pole and −55 ± 4% at the anodic pole. These values corresponded to membrane polarization by over 600 mV
Results, 3.4, paragraph 2; Figure 4BHow it will be judged
Di-8-ANEPPS, 1-us pulses, n = 5, averaged over all 60 pulse images; pole dF as in fig4a-fluovolt-limit; tolerances are the reported s.e. Measured pole IMP is dF / 9.1 x 100 mV. "Decreased below the calculated value" has no stated test; per cell the ratio of measured IMP to IMPC, and a one-sample t-test of the ratios against 1 (Methods 2.5), two- sided at p < 0.05. Reproduced when both maxima are within tolerance and occur at 570 V/cm, and 570 V/cm is the lowest strength at which the ratio is below 1 at p < 0.05 at one pole or more; partially reproduced when the maxima hold but the first significant strength is 780 V/cm (the Fig. 1 legend gives 570 to 780 V/cm).
- fig4b-first-10-pulses
Averaged over the first 10 of the 60 pulses, cathodic polarization reaches 66 +- 15% (710 +- 200 mV), so the 600-mV maximum over 60 pulses is not the dye limit.
Polarization measured this way reached 66 ± 15% (710 ± 200 mV) at the cathodic pole
Results, 3.4, paragraph 3; Figure 4B, dotted linesHow it will be judged
As fig4b-di8-threshold-570, with the polarization image collapsed from the first 10 of the 60 pulse images (Fig. 4B legend; Results 3.4). Tolerances are the reported s.e. Reproduced when both values are within tolerance and the first-10 maximum exceeds the 60-image maximum.
- fig4b-cathodic-pole-more-damaged
Above the threshold, pole IMP falls progressively as the field rises, more at the cathodic pole; at 1400 V/cm charging is no longer detectable at the cathodic pole and is about 30% dF (about -330 mV) at the anodic pole.
As the electric field increased further, the IMP reached at the poles progressively decreased, and the reduction was more pronounced at the cathodic pole ... At 1400 V/cm, charging was no longer detectable at the cathodic pole (i.e., the membrane became fully permeable) and dropped to ~30% ΔF (~ −330 mV) at the anodic pole
Results, 3.4, paragraph 4; Figure 4BHow it will be judged
Data as fig4b-di8-threshold-570. Reproduced when the mean |IMP| at each pole is lower at each strength above 780 V/cm than at the one below it, the drop from the pole's maximum to 1400 V/cm is larger at the cathodic pole, the cathodic dF at 1400 V/cm is not above zero by a one-sample t-test (two-sided, p < 0.05), and the anodic values are within tolerance. At 570 and 780 V/cm the paper says both poles are about equally permeabilized; the ratios to IMPC at the two poles are reported.
- fig4c-equator-follows-cosine
Membrane regions near the equator remain unaffected and follow the cosine dependence predicted by the Schwan equation up to 1000 V/cm, even when the poles are electroporated.
appeared relatively unaffected and followed the cosine dependence predicted by Schwan formula up to 1000 V/cm
Results, 3.4, paragraph 4; Figure 4CHow it will be judged
Data as fig4b-di8-threshold-570, the 360-point profiles. "Closer to the equator" fixes no width; readings, points within (a) 20 and (b) 40 degrees of the equator (both sides). Per strength, the mean over cells of measured IMP (dF / 9.1 x 100 mV) against IMPC at those points. A mean over points on both sides of the equator would cancel to about zero, so agreement is measured point by point - the slope of a least-squares line through the origin of measured IMP on IMPC over the equatorial points (the fit of Results 3.3). Reproduced when, under one width reading, at every strength up to 1000 V/cm that slope is within 0.9 to 1.1 (protocol default 10%, not the paper's) or its 95% interval includes 1, and at 780 and 1000 V/cm the pole ratio of measured IMP to IMPC is lower than the equatorial slope.
- fig4d-short-pulses-same-limit
With 0.3-us pulses of 357 to 3320 V/cm imaged at 275 ns, the maximum dF is 53 +- 4% (about 600 mV) at both poles, and the dependence on field strength is like that with 1-us pulses.
the maximum ΔF reached was 53 ± 4% (~600 mV) at both cell poles. The pattern of how the response changed with increasing PEF strength was not different from that with 1-μs PEFs either
Results, 3.4, paragraph 5; Figure 4DHow it will be judged
Di-8-ANEPPS, 0.3-us pulses, n = 10, image at 275 ns, IMPC at t = 275 ns; pole dF as in fig4b-di8-threshold-570; tolerance the reported s.e. Reproduced when both pole maxima are within tolerance and, as with 1-us pulses, |dF| at each pole decreases at the strongest fields after its maximum. The ratio of the 0.3-us to the 1-us maximum per pole is reported.
- fig5a-gd-extends-anodic-charging
1 mM Gd3+ extends anodic charging to -864 +- 44 mV against -542 +- 57 mV in controls (p < 0.001), and measured IMP stays nearly equal to calculated IMP down to -780 mV, so the dye response is linear at high IMPs.
1 mM Gd3+ extended the range of membrane charging at the anodic pole down to −864 ± 44 mV (p < 0.001 compared to −542 ± 57 mV in parallel control experiments, Fig. 5A). Furthermore, measured IMP remained nearly identical to the theoretical IMPC down to −780 mV
Results, 3.5, paragraph 1; Figure 5AHow it will be judged
Di-8-ANEPPS, 1-us pulses of 150 to 1400 V/cm, pole dF converted to IMP (9.1% per 100 mV), n = 7 Gd3+ and 5 control (Fig. 5A legend); tolerances the reported s.e.; test the 2-tailed t-test of the legend, unpaired. The extreme value is not defined; readings (a) per cell the most negative anodic IMP over strengths, averaged over cells; (b) the group mean at the strength where the group mean is most negative. Reproduced when, under one reading, both values are within tolerance and the groups differ at p < 0.001; and the Gd3+ group's anodic IMP is within 10% (protocol default) of its IMPC at every strength with IMPC down to -780 mV.
- fig5a-gd-cathodic-no-higher-but-no-decline
At the cathodic pole Gd3+ cells reach 647 +- 44 mV, not significantly more than controls, but Gd3+ prevents the progressive decline of IMP with stronger fields.
At the cathodic pole, polarization in the presence of Gd3+ reached 647 ± 44 mV, a value not significantly higher than that in the control group without Gd3+. However, Gd3+ prevented the progressive decline in IMP as the electric field was increased.
Results, 3.5, paragraph 2; Figure 5AHow it will be judged
Data and readings (a) and (b) of the extreme as in fig5a-gd-extends-anodic-charging. Part 1, a claim of absence on the paper's test - the Gd3+ cathodic maximum is not higher than the control's by the 2-tailed unpaired t-test at p < 0.05. Part 2 - the Fig. 5A legend's test; Gd3+ and control differ at p < 0.05 by 2-tailed t-test at the strongest strengths (those with IMPC above 1200 mV at the cathodic pole), with Gd3+ higher. Part 3 - the control's cathodic IMP falls from its maximum to 1400 V/cm and the Gd3+ group's falls less; the paper gives no measure for "prevented the decline", so the size of each fall and its s.e. are reported. Reproduced when the value is within tolerance and all three parts hold.
- fig5b-conductivity-calibration
Under Eq. (2), effective membrane conductivity barely affects charging while it stays far below cytosol and medium conductivity; a 10-fold increase reduces IMP by about 2% and a 100-fold increase by 12%.
its 10-fold increase would go unnoticed, yielding just a ~2% IMP reduction, and a 100-fold increase would reduce IMP by just 12%
Results, 3.5, paragraph 3; Figure 5BHow it will be judged
A computation from Eq. (2) with the Methods' parameters (li 0.4 S/m, lo 1.64 S/m, R 8 um, d 5 nm, intact lm 3e-7 S/m); it uses no recording. Reduction is 1 - f/1.5. The tolerance for "~2%" is half its last digit (0.5 percentage point), as the value is given to one significant figure; the 12% has the default 10%. Eq. (2) as read in the header gives 1.3% for 10-fold and 11.9% for 100-fold (1 - f/1.5), or 1.2% and 11.7% taken relative to the intact f; the "~2%" may be read from the Fig. 5B graph, and the equation itself is read from garbled text, so a mismatch at 10-fold is reported with both values.
- fig5c-gd-reduces-conductivity
For most datapoints, Gd3+ reduces the electroporation-induced effective membrane conductivity 2- to 3-fold.
For most datapoints, Gd3+ reduced the electroporation ... induced effective conductivity 2–3-fold
Results, 3.5, paragraph 4; Figure 5CHow it will be judged
For each datapoint of Fig. 5A (pole and field strength, group means of measured IMP and IMPC), lm,eff interpolated with Eq. (2) from the ratio of measured to calculated IMP (Fig. 5C legend); the fold is control lm,eff over Gd3+ lm,eff at the same pole and strength. Pairs where either ratio is 1 or above (no finite increase) are reported and left out. Reproduced when more than half of the remaining folds lie within 1.8 to 3.3 (the range widened by 10% of each bound).
- fig6a-ca-single-pulse
A single 1-us pulse causes clear Ca2+ entry at 1000 and 1200 V/cm, with the signal elevated through the 5-min observation; the small rise after 780 V/cm is not significant.
established a clear peak of Ca2+ entry at 1000 and 1200 ... and Ca2+ signal remained elevated through the 5-min observation period (Fig. 6A). A subtle ΔF increase after exposure at 780 V/cm was at the detection limit and not statistically significant.
Results, 3.6, paragraph 2; Figure 6AHow it will be judged
Fluo-4 dF in a circular region over the cell, background-subtracted, F0 the mean before the pulse (Methods 2.4); 14 cells per strength (Fig. 6A legend). The test and the time point are not given. Test readings, both used in Section 3.6 for Fig. 6B - (i) the one-sample t-test against zero, two-sided, p < 0.05; (ii) an unpaired t-test against the 0 V/cm group, two-sided, p < 0.05. Time point readings (a) the peak dF after the pulse; (b) dF at the end of the recording (the measure of Section 3.6 for Fig. 6B). Reproduced when, under one test and time reading, dF at 1000 and 1200 V/cm is above zero (or the 0 V/cm group) at p < 0.05, the end-of-recording dF at those strengths is above zero, and 780 V/cm is not significant under that same reading. The 780 V/cm part is a claim of absence on a test the paper does not name; every reading is shown, and when they give different outcomes for it the claim is methods_underspecified.
- fig6b-ca-train-thresholds
With trains of 60 1-us pulses at 6 Hz, a strong Ca2+ response occurs in 28% of cells at 780 V/cm and nearly all cells at higher strengths; at 650 V/cm only 2 cells (5.9%) respond strongly, but the mean signal at the end is above zero (p < 0.001) and above that at 230 V/cm (p < 0.05, one-tailed).
A strong Ca2+ response was recorded in 28% of cells exposed at 780 V/cm and in nearly all cells at higher intensities (Fig. 6B). At 650 V/cm, only 2 cells (5.9%) showed a strong response, but the mean Ca2+ signal by the end of recording was significantly different from zero (p < 0.001, one sample t-test) and marginally higher than after exposure at 230 V/cm (p < 0.05, single-tailed t-test)
Results, 3.6, paragraph 3; Figure 6BHow it will be judged
Fluo-4 dF as in fig6a-ca-single-pulse, 28 to 54 cells per strength, 224 in all, each tested once. The mean-signal tests are fixed by the text - dF at the end of the recording (read as the last frame; the mean of the last five frames, 10 s, is shown as a variant), one-sample t-test against zero, and an unpaired one-tailed t-test against 230 V/cm. "Strong response" is not defined in the paper, its legends or Methods; no threshold can be taken from them, so the two percentages are judged only if the dataset marks responders, and otherwise are blocked as methods_underspecified with the distribution of end dF per strength shown. Reproduced when the two tests reach p < 0.001 and p < 0.05 (and the percentages match where they can be judged).
Pipeline
- Paper
- done (1 of 1)
- Claims
- done
- Claims check
- done (ready)
- Attempt
- todo
- Questions
- none
- Verified
- none