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In recent years the jellyfish Aequorea victoria has provided the scientific community with a pair of tools of exceptional usefulness: aequorin and the green fluorescent protein (GFP). Whereas the former has played a major role in the study of calcium signalling, the latter has sparked the imagination of researchers into a myriad of elegant experiments. The firefly Photinus pyralis has also been of great use, providing a third luminescent protein, luciferase, which is mostly known for its role as a reporter protein. Concurrent use of these three proteins provides a powerful means of elucidating biological processes with fine spatio-temporal detail. Here we will illustrate how specific molecular engineering of these three proteins provided a set of biological tools capable of generating important data in the field of calcium homeostasis. First, we will show how the use of specifically targeted aequorin chimeras enabled the measurement of regional Ca(2+) concentrations; second, how the use of GFP (and derived chromatic mutants) permitted detailed morphological analyses in living cells; third, how luciferase was used to analyse energetic requirements at the subcellular level. Together, these three experimental approaches have provided important details on how mitochondria participate actively in calcium homeostasis. A final note regarding clinical implications demonstrates the practical usefulness of the data obtained.