Journal: Frontiers in Molecular Neuroscience
Article Title: There is no F in APC: Using physiological fluoride-free solutions for high throughput automated patch clamp experiments
doi: 10.3389/fnmol.2022.982316
Figure Lengend Snippet: HEK and CHO cells recorded in standard and fluoride-free internal solution. (A) HEK cells expressing hERG were captured with almost 80% success rate for R Seal ≥ 1 GΩ + I Tail > 150 pA and almost 40% success rate for R Seal ≥ 1 GΩ in fluoride-free internal solution (left). If a QC cutoff of R Seal ≥ 0.25 GΩ + I Tail > 150 pA was used, the success rate was increased in both conditions (right). (B) CHO cells expressing Na V 1.5 were captured with almost 80% success rate for R Seal ≥ 1 GΩ + I Peak < –100 pA and over 40% success rate for R Seal ≥ 1 GΩ in fluoride-free internal solution (left). If a QC cutoff of R Seal ≥ 0.25 GΩ + I Peak < –100 pA was used, the success rate was increased in both conditions (right). (C) Histogram plot of the R Seal values for HEK-hERG cells in standard and fluoride-free internal solution. For R Seal > 4 GΩ, although they are high giga-seals, the seal resistance is not accurately calculated and is therefore shown as an extra bar and these R Seal values were not included in the calculation for R Seal . (D) Histogram plot of the R Seal values for CHO-Na V 1.5 cells in standard and fluoride-free internal. For R Seal > 4 GΩ, although they are high giga-seals, the seal resistance is not accurately calculated and is therefore shown as an extra bar and these R Seal values were not included in the calculation for R Seal .
Article Snippet: We demonstrate this using the ion channels Na V 1.5 and Na V 1.7 expressed in CHO (Charles River), hERG expressed in HEK293 (SB Drug Discovery), and K Ca 3.1 (SK4) also expressed in CHO cells (Charles River).
Techniques: Expressing
