Acquisition of a Laser Microdissection Microscope with a Special Cooling Chamber and an Associated Cryostat

The GLOBE Institute comprises a wide range of expertise and skills within its research programs spanning modern and ancient genomics, macroecology, geobiology and astronomy.

A fully automated laser microdissection microscope equipped with an especially designed cooling chamber will enable us to generate georeferenced microsamples that will be used to generate three-dimensional microreconstructions of microbial communities and adjacent host tissues in animal intestines. The system is moving the laser, not the sample, which is extremely important for our setup. This laser will cut along the masked area of interest. It will be equipped with a variety of collecting devices, like strips of 8 wells, 96 well plate, 96 micro tube plate on a custom stage and the system is collecting the dissected samples via gravity, which is crucial for our workflow. Those dissected samples will then be used for DNA and RNA sequencing, which will require slides to be kept as cold as possible. To that aim, the microscope will be equipped with a novel prototype cooling chamber designed specifically for us to ensure slides are kept at < 5ºC.

— leica LMD7 moves the laser, not the sample (stage independent — high precision and speed);

— cuts directly, real-time with ‘Move and Cut’;

— specimen collection by gravity. This prevents any possible contamination and stress to the sample during the removal phase;

— collects samples into strips of 8 wells, 96 well plate, 96 micro tube plate on a custom stage for our system;

— LMD7 is the only one to use a solid-state UV laser pulse with a wavelength of 349 nm, whereby one can adjust laser power and aperture directly during the laser cut;

— our research requires the stage to be at near 0 ºC to avoid decay of DNA and RNA, and to be inside a closed chamber to minimise contamination. We have worked on that solution for over a year with Leica and its subcontractor Okolab, and they are the only ones who can deliver such a solution for us;

— we are collaborating with Leica on developing new features for this microscope, that are needed for our experimental workflow. This is crucial for moving a step further with our research and therefore it is crucial to purchase the very same microscope;

— in Leica's cryostats, incl. the CM3050 S, you have a ‘One block chamber’ which means a completely enclosed chamber where all parts are welded together, so no waste can disappear into ‘the unknown’. Therefore this cryostat provides a system that is very easy to clean and maintain, when handling either fresh tissue, or when limiting contamination is an important factor;

— the chamber temperature goes down to -40 ºC (Many other cryostats have -35 ºC). It is important to keep as low temperature to prevent degradation of nucleic acids;

— programmable reverse counter function;

— a custom prototype cooling chamber designed specifically for our purposes will be implemented.

Patent numbers

— EP1276586B1;

— DE10018251C2;

— DE10057292C2;

— EP1207392B1;

— DE102005028062C5;

— EP1893971;

— EP1719998B1;

— DE102009029078;

— DE102012218382;

— DE102014202860.