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The study mainly involves sensor adjustment, probe immobilization and ice bath sample addition (at different concentrations and different volumes).
For sample preparation, the etching bath sample is diluted before analysis due to the high-HF concentration.
First, the pH of the etching bath sample is adjusted to pH 6.5 to 7.5 with sodium hydroxide solution (1.0 mol L−1).
For sample preparation, a buffer solution (TISAB II from Sigma-Aldrich Chemie GmbH, Steinheim, Germany) is added to the diluted etching bath sample.
We follow the difference method of Hach Lange [8]: The etching bath sample is diluted in order to fit to the measurement range and added to the decomposition cuvette.
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On each day of experiments, bath samples were accompanied by a set of standards and processed similarly.
Transmural MS rates were expressed as ρmol/cm× min after correction of periodic bath samples for total bath volume.
Due to the different expected concentrations of the target compounds in cytosolic and bath samples, different calibration points were used.
The concentrations of the analytes were finally calculated as pmol/mg wet tissue for both the cytosolic and the bath samples.
Using a dry ice/ethanol bath, samples were freeze-thawed twice followed by DNAse I treament for 20 min at room temperature.
For bath samples, a seven-point calibration curve in range of 0.05 to 10 μM was used with injection volume of 30 μl.
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