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Biology

Fluorescence microscopy offers the wonderful opportunity to probe biological systems and to map biochemical events in the living cell. Nowadays, we can glimpse at the functioning of molecular machineries and cell signalling, and we can try to link molecular properties to cell and tissue phenotype. This is permitting us to have a better understanding of the fundamental principles in biology and to dissect the molecular events that underlie disease.

Because of their impact to society, we have been studying the molecular patho-physiology of Parkinson’s disease (ENI, UCAM), malaria (UCAM) and cancer (MRC, UU). We (MRC) are currently engaging in an extensive research program to enable optical biophysical techniques for the study of the systems biology of cancer.

On the right: fluorescence lifetime imaging of malaria-infected red blood cells loaded with the fluorophore calcein

Biotechnology

Molecular biology and engineering of fluorescent proteins provide powerful tools to probe cellular physiology and the molecular mechanisms of disease. A variety of fluorescent genetically encodable biosensors have been developed in the recent years to detect calcium concentration (cameleons, camgaroos and pericams), chloride (clomeleon), cell redox state (rxyFP), kinase activity (AKA, ATOMIC, Erkus, Picchu, ...) and pH (phluorins and deGFP). Based on the design of CY11.5 tandem protein developed by Miyawaki’s groups and CY11.5’s pH sensitivity, we (ENI) developed a family of FRET based biosensors for pH: pHlameleons.

At the Hutchison/MRC Research Centre in Cambridge we are now developing an innovative sensor platform for multiplexing a variety of kinase reporters with high spatial and temporal resolution.

On the right: fluorescence lifetime of pHlameleon beads for sensing pH (shown pH 6, 8 and 10)

Technology

New horizons in biology and medicine are opened by technologies that allow the investigation of molecular mechanisms which regulate cell/tissue function and disfunction. We (ENI, UU, UCAM, MRC) aim to develop novel integrated detection systems for quantitative detection of emission spectra, fluorescence lifetime and polarization. New techniques and technologies have to be user-friendly and cost-effective in order to allow non-specialist biomedical laboratories to take advantage of biophysical imaging. Also, cost-effective systems will permit the implementation of new tools for drug-discovery and diagnostics.

On the right: a CCD/CMOS camera for wide-field FLIM.

Theory

Theoretical work is necessary for the proper interpretation of data, for modelling biological systems, for the understanding of the physical phenomena exploited for sensing applications and for engineering the optimal detection systems.

Fundamental questions that can be addressed with theoretical works are, for instance: which is the information content of images in fluorescence microscopy? How many photons are required to correctly estimate a fluorescence lifetime? Also, we (MRC) aim to push biophysical imaging techniques to characterize the systems biology of cancer: how protein networks code for diverse functions?

On the right: Jablonski diagram of a FRET pair and fluorescence lifetime image of a cell expressing fluorescent proteins undergoing energy transfer


The old pages of Quantitative-Microscopy.Org have been reorganized under two domains:

  • this site: pages for the dissemination of my research activities and, particularly, of the projects supported by funding bodies
  • WikiScope.Org an educational outreach about all type of microscopy



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