Stimulated Raman Spectroscopy: A new tool for profiling intracellular catalysis

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Understanding how fast small molecules (such as drugs) or biomolecules (such as DNA) react inside cells is crucial to the development of new medicines and the design of successful industrial processes which rely on bio-transformations. And yet at the moment there are no tools which can give both the location of a biomolecule and its rate of reaction in live cells simply relying on the inherent properties of that molecule. Current strategies either require the cell or tissue under examination to be fixed (as for histological slides) so that the concentrations of compounds can be mapped by mass spectrometry (thus not applicable for examining reactions in living cells), or rely on the incorporation of isotopes to allow detection of the molecule under study by its nuclear magnetic resonance (a technique which does not allow the position of reacting molecule within the cell to be accurately determined).A new technique, called stimulated Raman spectroscopy (SRS), allows the vibrational properties of a molecule to be measured in a way that is:* quantitative - allowing accurate measurement of local concentrations;* spatially resolved - allowing molecules to be mapped inside cells;* chemically discriminating - allowing different types of molecules to be imaged at the same time;* and uses non-destructive optical light for detection - meaning that the studies could eventually be transferred to fibre optic technologies to be used in patients.We propose to use SRS to study the rate of one particular reaction inside cells so that we can demonstrate that it is possible to do this both in the presence of all of the other signals from the cell, and in a way that allows us to show exactly where the reacting molecules are inside a cell.