When studying the toxic effects of different compounds on living cells, it may be challenging to find the biologically correct moment for measurements. However, carrying out measurements at precisely the right moment is essential, because the non-optimal growth stage of cells may lead to erroneous results.
Experimental arrangements
Tecan’s application note “Walk-Away Monitoring of Cytotoxicity, Viability and Apoptosis” describes an experiment in which cytotoxicity and viability in living cells were monitored in an automated manner using the Spark microplate reader. The experiment used the device’s ability to automatically inject the samples with the caspase-3/7 reagent when a specific level of cytotoxicity was exceeded.
The experiment also evaluated the impact of six kinase inhibitors on the viability of K562 cells continuously for 72 hours. The environmental control properties and automatic lid opener of the microplate reader were utilised in the experimental arrangements.
The results
It was observed during the experiment that a long, automated kinetic analysis enables finding the correct moment for measuring cytotoxicity, making it easier to study the functional mechanisms of compounds. According to the preliminary results, changes in the cell membrane correlate with the observed level of apoptosis. In addition, it was found that cytotoxicity measurement can be used to determine the correct moment for adding the caspase-3/7 reagent in studies. The threshold value for caspase-3/7 activity, on the other hand, correlated with compounds known to induce apoptosis.
The original application note can be downloaded here:
Tecan application note: Walk-Away Monitoring of Cytotoxicity, Viability and Apoptosis