Introduction

An efficient method of systematically analyzing brain functions requires simultaneous monitoring of the electrical activity of many cells in a given brain volume. The STED (Stimulated Emission Depletion) microscopy scanning system allows, respectively, the study of high-speed phenomena and/or the visualization of nano-structured objects, exceeding the diffraction limits given by Multi-Fotonic Microscopy (MP). In addition, it is able to swiftly steer the focus of the two beams of light on a plurality of predefined positions within a given volume.

Technical features

Several technologies have been developed to make rapid three-dimensional measurements on the brain tissue, one of these are the acousto-optical deflectors (AOD), used to perform random access imaging and quickly vary the focus of a laser beam without mechanical movements. Currently, these and other objects are included in the STED microscopy optical scanning system of the present invention, consisting of: 1.) an optical system to guide a first and a second beam of light, 2.) a means of deflection to divert these first and second beams of light in a variable direction, in which these means of deflection include at least one AOD. According to the invention, the two beams of light enter the random access scanning system from opposite sides and propagate through each AOD element. Depending on the number of AOD elements it is also possible to provide a system that can scan in multiple directions: with a single AOD you can perform a one-dimensional scan, while with 4 AOD you can perform a three-dimensional scan.

Possible Applications

• Field of fluorescence microscopy and brain tissue functional imaging
• Nanolithography, nanomanufacturing
• Research and storage of optical information and other fields involving high-speed phenomena

Advantages

• Overcoming the diffraction limitation of MP microscopes
• Higher spatial resolution
• Reliable, economical and easy to implement