BX-026 Enejder - Gustafsson Imaging of Glucose in Living Cells by CARS-microscopy
Project Summary: Objectives, expected results, summary of the Project plan
Presently, there are no imaging methods available to visualise the trafficking, intracellular localisation and metabolism of low molecular metabolites in vivo on a single-cell level. We have identified a novel microscopy technique based on the Coherent Ant-Stokes Raman Scattering (CARS) process with a high potential to fulfil this need. It will allow exclusive insight into one of the most central processes in life in its systemic context, of importance for a wide range of fields, not the least for medicine, pharmacology and biotechnology. CARS microscopy offers the unique possibility to image molecules based on their characteristic vibrational properties, without the need for any reporter molecules. This allows direct imaging of small molecules, such as glucose, not possible with other microscopy modalities. For imaging with improved chemical specificity, we intend to deuterate glucose and probe the unique C-D bond, vibrating at a frequency far from other bio-molecules. This novel deuteration-approach may result in the final establishment of CARS microscopy as a significant technique within applied cell biology. Project plan: (1) Extra- and intracellular glucose concentrations will be monitored in living cells, to visualise, quantify and model overall transport and regulatory mechanisms. Yeast cells will be used as model organism. (2) Fluxes and pools of glucose intermediates will be imaged with parallel monitoring of signalling protein cascades. This type of multi-parametric study is essential, as glucose signalling is intertwined with a complex array of other signalling pathways. (3) Finally, sub-cellular localisation of these processes (to the nucleus, mitochondria, vacuoles and other cellular membrane-enclosed compartments), again in parallel with timeresolved monitoring of signalling, regulatory and catalytic proteins, will bring biology to hitherto unknown levels.
Keywords for the project (to be found in international databases)
Bio-imaging, non-linear microscopy, glucose, metabolism, regulation, signalling, single-cell detection, CARS