Abstract
Although many laboratory experiments are available to illustrate spectrophotometric or fluorometric methods, few of them introduce the use of luminometry to students. Bioluminescence, a subtype of chemiluminescence, is produced when an enzyme-catalyzed chemical reaction gives rise to light emission. Despite the advantages of bioluminescent methods, including sensitivity and specificity, and their increasing use in experimental sciences and biomedical laboratories, their presence in courses is almost nonexistent. The luciferase-catalyzed enzymatic reaction has generated a myriad of practical applications, including those derived from the measurement of the ATP consumed in the reaction. In particular, the measurement of ATP levels in drinking or stored waters directly correlates with their bacteria content, facilitating the development of rapid methods for detecting bacterial contamination. This avoids the long waiting time associated with traditional microbiological methods, based on the growth of the microorganisms in a suitable culture medium. Over the past two years at the University of Malaga, we have implemented a new laboratory experiment for undergraduate chemistry and biochemistry students. In this experiment, students detected bacterial contamination in water by quantifying ATP with the luciferase-catalyzed reaction. The experiment was successfully implemented in two different formats, either as a full project developed by students throughout the entire duration of the academic course, or as a short protocol, carried out in a single laboratory session. Between them, a whole range of intermediate options could be arranged by educators to suit their course requirements and the learning objectives to be achieved by students.
Highlights
Index of the topics covered in the luminometry lecture (.pdf)
In both formats used, either as a project developed by students throughout the full course or as a short protocol carried out in a single laboratory session, the students’ achievement of the learning goals was very satisfactory
Summary
There are many laboratory experiments available to illustrate spectrophotometric or fluorometric methods, few of them introduce the use of luminometry to students. By measuring ATP concentrations in water, students gain knowledge about the principles and 110 applications of bioluminescence and the practical use of luminometers This is already valuable in itself because, as mentioned, luminometry is a neglected technique in our classrooms, in terms of laboratory experiments availability. The learning goals for the PBL format include those from the short protocol, and others derived from the more active role played by students in their learning process, making them undertake authentic real-world tasks, similar to those they will find in 115 their professional future Many of these students will face for the first time common issues for a professional, such as the acquisition of reagents and materials, the assessment of the necessary instruments and equipment, the adaptation and scaling of experimental protocols, or the analysis of costs and operational feasibility, among others
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