Correlative Fluorescence Microscopy and Advanced Low-energy ESEM for Imaging of Immuno- Labelled Wet Biological Samples

The grant deals with basic research of electron-gas interactions for observation of highly sensitive wet immuno-labelled plant cells at low electron beam energies. This will enable the realization of new concept of correlative fluorescence microscopy and advanced low-energy environmental scanning electron microscopy (LEESEM) for studies of immunostained plasma membrane protein complexes in the tobacco cells. A new strategy for complex simulations of electron interactions with a precisely defined working gas mixture in dynamically changed conditions of temperature, pressure, flow rate (ANSYS Fluent) including emission of signal electrons from the sample (GEANT4) will be implemented using improved Monte Carlo software. New detection systems will be designed to validate the simulation results. These will enable a higher resolution and field of view of LEESEM. Using the Thermodynamically Optimized Low Temperature Method, microstructural modifications of samples during observations in LEESEM will be minimized, which is crucial for the correlation of images of both methods.