Elisa Zagato

Elisa Zagato


Elisa Zagato
PhD student, Biophisicist
Laboratory for General Biochemistry and Physical Pharmacy
Ghent University
Harelbekestraat 72
9000 Gent
Tel: 0032 9 264 8047 (secretary)
Tel: 0032 9 264 8095 (direct)
Fax: 0032 9 264 8189
E-mail: elzagato.Zagato@UGent.be



Master in Physics



Research interests:

light sheet microscopy

Summary of Research Project(s)

Disposable sample holder with integrated optics for enabling light sheet microscopy on standard light microscopes

Optical microscopy, and especially fluorescence microscopy, has seen a range of important developments in recent years that are linked to novel illumination strategies and optical probes. Fluorescence super-resolution techniques like Stimulated emission depletion microscopy (STED) and Stochastic Optical Reconstruction Microscopy (STORM) are notable examples. Another important development is Light Sheet Microscopy (LSM) that has emerged as a powerful alternative to confocal scanning microscopes. LSM uses a thin sheet of light that illuminates a particular plane in the sample orthogonal to the detection pathway. By letting the light sheet coincide with the focal plane of the detection objective lens, only objects within the focal plane are illuminated, while regions outside the focal plane remain dark. This results in an optical section with excellent contrast due to the absence of out-of-focus light. Compared to classic confocal microscopy, LSM has the advantage of offering high-speed imaging with superior detection sensitivity at a substantially reduced cost.

Despite a number of remarkable successes, LSM has not found widespread application thus far. The major difficulty in LSM is the necessity of positioning two bulky objective lenses (for illumination and imaging) at a 90° angle in close proximity with the sample. Such a configuration is not compatible with commercial microscope bodies which are designed for a single objective lens. Thus, LSM has only been realized in a few specialized groups who have developed their own optical system and custom made sample chamber. In this project the aim is to address this problem by developing two types of sample holders with microfabricated integrated optical components for illumination so as to enable LSM on standard light microscopes. Special attention goes to a cost-effective design which would allow the sample holders to be used as disposables, an important asset when working with biological samples.

The potential of the LSM sample holders will first be demonstrated for improved nanoparticle characterization. In our laboratory it was recently shown that Single Particle Tracking (SPT) microscopy allows to accurately measure the size distribution and concentration of nanoparticles in solution. The increased signal to background ratio offered by the LSM chips is expected to extend the working range of SPT to smaller nanoparticles and higher concentrations. Accurate size and concentration measurements are of importance to a wide diversity of nanoparticle related fields, including pharmacy (nanomedicines), biomedicine (nanoparticle based imaging, therapies and diagnostics) and nanotoxicology (e.g. detection of environmental nanoparticles).

Secondly, the usefulness of a particular LSM sample holders design will be assessed for live cell imaging applications. As a case study, the intracellular trafficking of nanomedicines for gene therapy in living cells will be imaged using the LSM sample holder. Although intracellular trafficking of nanoparticles was recently demonstrated in our laborastory using a regular epi-fluorescence microscope, due to the limited signal to background ratio, particles could not be tracked in regions of the cell with a high background, thus resulting in a loss of information. It is expected that the improved signal to background ratio offered by the LSM sample holders will improve the visibility and tracking of nanoparticles in cells.