The Silver Lab
UCL logo

Microscopy development and application

A primary objective of our lab is to understand how the brain performs parallel distributed signal processing. However, investigating information processing in the brain has been severely hampered by the limited experimental methods available for studying rapid signalling in 3D structures. Functional optical imaging using 2-photon microscopy (Denk et al. 1990) has been increasingly used to investigate neuronal signalling, because it allows submicron resolution and good tissue penetration. However, conventional 2-photon imaging using galvanometers is too slow to capture the timing of fast neuronal signalling events and, as for most optical microscopes, is limited to 2D imaging. To overcome these limitations we have developed a new type of optical element based on 4 acousto-optic deflectors that can focus and steer a laser beam at ~40 kHz. The concept of a fast acousto-optic lens (AOL), which was first proposed by Kaplan (Kaplan et al. 2001) is shown in Figure 1. Our lab (Kirkby et al. 2010) as well as Prof. Peter Saggau's lab (Duemani Reddy et al. 2008) have developed AOLs for use with 2-photon microscopy. This high speed 3D functional optical imaging technology enables many points of interest to be measured at kHz rates and is well suited for measuring distributed neuronal signalling.

Our 3D 2-photon AOL microscope (Figure 2) has many new design features that overcome several of the technical limitations of this technology. This has enabled a 10-fold reduction in the laser power required for imaging and an extended focussing depth range. These features now make it useful for neuroscience research applications. The image in Figure 3 shows a cortical neuron and its dendritic processes filled with a fluorescent dye. It illustrates the submicron resolution achievable when the AOL is used to focus over 100 microns. This 2-photon AOL microscopy also provides a new way to image signals in neural populations distributed in 3D space. Our preliminary in vitro and in vivo data, using calcium sensitive fluorescent dyes to monitor neural activity, shows that signalling can be monitored simultaneously from more than 30 neurons distributed in 3D space with a temporal resolution greater than 1 kHz. This time scale is relevant for 'reading' spike timing-based neural population codes.

UCL Business has filed two patent applications on this technology is hoping to licence the design features of our 3D 2-photon AOL microscope. This technology has several potential applications outside neuroscience and cell biology. These include high capacity data storage, 3D lithography, laser tweezers and cold atom traps for quantum physics experiments.












This page last modified 28 March 2011
University College London - Gower Street - London - WC1E 6BT - Telephone: +44 (0)20 7679 2013 - Copyright © 1999-2011 UCL
Disclaimer | Accessibility | Privacy | Advanced Search | Help



Search by Google