Abstract:
Luciferase reporter assays represent a simple and sensitive experimental system in cell and molecular biology to study multiple biological processes. However, the application of these assays is often limited by the costs of conventional luminometer instruments and the versatility of their use in different experimental conditions. Therefore, we aimed to develop a small, affordable luminometer allowing continuous measurement of luciferase activity, designed for inclusion into various kinds of tissue culture incubators. Here, we introduce LuminoCell-an open-source platform for the construction of an affordable, sensitive, and portable luminometer capable of real-time monitoring in-cell luciferase activity. The LuminoCell costs $40, requires less than 1 h to assemble, and it is capable of performing real-time sensitive detection of both magnitude and duration of the activity of major signalling pathways in cell cultures, including receptor tyrosine kinases (EGF and FGF), WNT/β-catenin, and NF-κB. In addition, we show that the LuminoCell is suitable to be used in cytotoxicity assays as well as for monitoring periodic circadian gene expression.
Authors:
Kamila Weissová 1, Bohumil Fafílek 2 3 4, Tomasz Radaszkiewicz 5, Canan Celiker 1, Petra Macháčková 6, Tamara Čechová 3 4, Jana Šebestíková 1 5, Aleš Hampl 1 4, Vítězslav Bryja 5, Pavel Krejčí 2 3 4, Tomáš Bárta 7 5
Affiliations:
1 Department of Histology and Embryology, Faculty of Medicine, Masaryk University, Brno, Czech Republic.
2 Institute of Animal Physiology and Genetics of the Czech Academy of Sciences, Brno, Czech Republic.
3 Department of Biology, Faculty of Medicine, Masaryk University, Brno, Czech Republic.
4 International Clinical Research Center, St. Anne’s University Hospital, Brno, Czech Republic.
5 Department of Experimental Biology, Faculty of Science, Masaryk University, Brno, Czech Republic.
6 Cellular Imaging Core Facility, Central European Institute of Technology (CEITEC), Masaryk University, Brno, Czech Republic.
7 Department of Histology and Embryology, Faculty of Medicine, Masaryk University, Brno, Czech Republic tbarta@med.muni.cz.
