PARAFLUO

Parallel fluorescence spectroscopy tools for micro and nano-analytical applications down to single biomolecules

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 Obiettivo del progetto (Objective)

'Analytical methods based on fluorescence measurements are widely employed for investigating biological process at cellular level. A modern technique is fluorescence-lifetime imaging microscopy (FLIM), where a map is obtained of the fluorescent emission lifetime versus position in a cell. The objective of project PARAFLUO is an innovative instrumentation system that will enhance and extend the usefulness of FLIM, making possible to obtain simultaneously FLIM data separately for the various spectral components of the emission. There is wide consensus among experimenters that this spectrally resolved technique (called sFLIM) will support a better understanding of the biological processes involved. Such understanding is paramount for the (patho)physiology of tissues and organisms and gives a base for gaining a better insight in key medical issues, such as the origin and growth mechanisms of tumors. The optoelectronic instrumentation developed will be useful also for other market objectives, such as simultaneous multi-spectral profiling of objects by laser detection and ranging (LADAR) techniques. The developments envisaged are essentially: (a) a photon-counting array detector based on the silicon single-photon avalanche diode (SPAD) technology; (b) a new micro-lens system for focusing light onto the detector and (c) an ASIC based multichannel time correlated single photon counting (TCSPC) system, integrated with an optoelectronic setup in a confocal microscope. The base of the PARAFLUO consortium is given by three SME’s; each one having a consolidated technical know-how and an active presence in the market over one of the quoted scientific-technical (S/T) areas. Five RTD performers have been selected primarily because of their high international standard in these areas; furthermore, each of them has experience of active collaboration with the SME directly concerned by the specific S/T work. A professional partner supports the coordinator and ensures timely and efficient exchange of materials and information in the project.'

Introduzione (Teaser)

Analytical techniques that provide information on the interactions among biomolecules could help discover the micro- and nanoworlds of biological processes.

Descrizione progetto (Article)

The majority of biological processes entail a complex interplay among different macromolecules such as proteins, sugars, lipids and nucleotides. To understand and monitor these processes and their impact on tissue physiology, scientists have employed fluorescence imaging techniques.

Fluorescence-lifetime imaging microscopy (FLIM) is a technique that relies on the decay of fluorescence measuring the changes either of the spectral intensity or of the lifetime of the fluorescence. As a result, it could be used to discriminate among different environments and to provide information about the labelled molecules and their interactions.

Based on this, the EU-funded http://www.parafluo.com/ (PARAFLUO) project aimed to address the rising quest for analytical instrumentation suitable in this field. To this end, project partners set out to develop optoelectronic instrumentation capable of carrying out spectrally resolved FLIM.

Project activities included the design of a photon-counting array detector based on the silicon single-photon avalanche diode technology. Additionally, a microlens system was fabricated and integrated onto the detector alongside a multi-channel time-correlated single-photon counting system. The whole setup was attached to a confocal microscope to facilitate visualisation of labelled cells or tissues.

The advanced optoelectronic instrumentation generated by PARAFLUO small businesses will enhance their competitiveness in the European market. From a research viewpoint, it will help to unravel the molecular machines that orchestrate cellular processes and to gain better insight into key medical issues.