Researchers at the Max Planck Institute of Biochemistry have used ultrasensitive scientific digital cameras to improve understanding of how enzymes can regulate diverse cellular processes. Their basic research could eventually benefit researchers developing anti-cancer treatments and drugs to prevent transplant rejection.
Doris Berchtold and Tobias Walther from the Max Planck Institute of Biochemistry used a Revolution microscopy system from Andor Technology with an Andor iXonEM+ 897 back-illuminated EMCCD in their work with Target of Rapamycin (TOR) kinases, a type of enzyme that act on proteins to coordinate cell growth and division.
In their active form, the enzymes are assembled together with other proteins into two complexes – TORC1 and TORC2. TORC1 responds to rapamycin, an immunosuppressant drug used to prevent organ transplant rejection. The role of TORC2 is less clear but it may help regulate the position of cell growth. The microscopy system and EMCCD are used to investigate where TORC1 and TORC2 are found within a cell, with an aim to understand the enzyme's different biological function and responsiveness to rapamycin.
The researchers imaged fluorescent labels attached to components of the TOR complexes in yeast cells. Berchtold and Walther found TORC1 and TORC2 located in different, spatially-separated cellular compartments, with TORC2 acting in a previously unrecognised plasma membrane domain.
'For ten years, scientists have tried to localise these kinases indirectly,' said Walther. 'But, with this new generation of cameras, we can see single molecules in living cells – something which was impossible before.'
Walther gives credit to Andor's Revolution laser spinning disk system, which the company supplied to the lab's specific requirements and includes photo-bleaching and activation capability.
'My lab does a lot of microscopy and for all of our applications, these new cameras have been a breakthrough. But it's not just the cameras,' he said. 'It's also the coordination of all parts of the Revolution system to achieve our goals that makes the difference - the little things that people don’t think much about until they use them.'
