Fluorescence lifetime imaging is a rather new and effective tool
that can be used to study complex biological samples, either at microscopic or
macroscopic levels. The map of the fluorescence lifetime allows one to
discriminate amongst different fluorophores and to achieve valuable insights
into the behaviour of emitting molecules, leading to information like local
pH, oxygen concentration in cells, etc. Moreover, the distribution in space
of any fluorescent marker achievable with this technique can be exploited
for diagnostic purposes in medicine. After a brief introduction on the
motivations for applying fluorescence lifetime imaging in biology and
medicine, the basic principles of this technique will be addressed. Then,
the two possible implementations of fluorescence lifetime imaging (i.e. the
frequency domain and the time domain methods) will be presented. For this
purpose, special attention will be devoted to practical aspects of image
acquisition and processing, especially for what concerns the time domain
method. Then, the analysis of the state-of-the-art systems will include a
brief discussion on new concepts that have recently been introduced in this
research field. Finally, two interesting applications of fluorescence
lifetime imaging will be presented. The former refers to skin tumour
detection and has been successfully applied in a preliminary clinical trial,
the latter regards DNA chips reading and has been tested only at laboratory
level, yet it has produced promising results for its future implementation in
commercial systems.