Time-Resolved Cell Culture Assay Analyser (TReCCA Analyser) for the Analysis of On-Line Data: Data Integration—Sensor Correction—Time-Resolved IC50 Determination

Abstract

Time-resolved cell culture assays circumvent the need to set arbitrary end-points and reveal the dynamics of quality controlled experiments. However, they lead to the generation of large data sets, which can represent a complexity barrier to their use. We therefore developed the Time-Resolved Cell Culture Assay (TReCCA) Analyser program to perform standard cell assay analyses efficiently and make sophisticated in-depth analyses easily available. The functions of the program include data normalising and averaging, as well as smoothing and slope calculation, pin-pointing exact change time points. A time-resolved IC50/EC50 calculation provides a better understanding of drug toxicity over time and a more accurate drug to drug comparison. Finally the logarithmic sensor recalibration function, for sensors with an exponential calibration curve, homogenises the sensor output and enables the detection of low-scale changes. To illustrate the capabilities of the TReCCA Analyser, we performed on-line monitoring of dissolved oxygen in the culture media of the breast cancer cell line MCF-7 treated with different concentrations of the anti-cancer drug Cisplatin. The TReCCA Analyser is freely available at www.uni-heidelberg.de/fakultaeten/biowissenschaften/ipmb/biologie/woelfl/Research.html. By introducing the program, we hope to encourage more systematic use of time-resolved assays and lead researchers to fully exploit their data.

Essential role of mitochondrial Stat3 in p38MAPK mediated apoptosis under oxidative stress

“…In good agreement to published findings 13 , 15 , we found less mitochondrial activity in cKD cells than in NC cells by measuring the oxygen consumption (SI. 10) 36 . We performed MTT assay (Fig.…”

“…Excitation and emission settings were 488 nm, 515–545 nm (FL1 channel) for JC monomers, and 564–606 nm (FL2 channel) for JC aggregates 23 . The oxygen consumption measurement was performed as our previously reported 36 .…”

Pharmacological activation of pyruvate kinase M2 reprograms glycolysis leading to TXNIP depletion and AMPK activation in breast cancer cells

“…Cells were seeded at a concentration of 150,000 cells/well in OxoDish and HydroDish 24-well plates (PreSens Precision Sensing GmbH, Regensburg, Germany) and incubated in standard conditions. As described previously [ 23 ], fluorescence-based sensors embedded at the bottom of each well could be read continuously using SensorDish Readers (SDRs) placed inside a standard incubator. The indicated treatments were added to each well; subsequently, the plates were mounted on readers inside the standard incubator, and medium pH values/oxygen saturation values were recorded each 10 min.…”

Expression of TXNIP in Cancer Cells and Regulation by 1,25(OH)2D3: Is It Really the Vitamin D3 Upregulated Protein?

“…OxoDish and HydroDish 24-well plates (PreSens Precision Sensing GmbH, Regensburg, Germany) were used for on-line measurements of dissolved oxygen and the pH of the medium, respectively, as previously described [ 29 ]. Changes in dissolved oxygen reflect oxygen consumption by respiration and hence mitochondrial activity, whereas changes in medium pH indicate lactate production from glycolytic metabolism.…”