Nonlinear optical microscopy techniques (such as two-photon florescence) are being used to acquire volumetric (i.e. three dimensional) images that probe several hundred microns into scattering tissue. These techniques are being combined with fast acquisition schemes to allow imaging of live, moving specimens at high NA. This combination allows for high-resolution, in vivo exploration of biological specimens. However, the added system complexity prevents these microcopy techniques from being readily exploited by the biological and medical communities that would greatly benefit from them. Therefore, we propose the development of a simplified confocal microscope configuration that rapidly acquires volumetric images within a scattering media. It uses cost-effective and user-friendly components, meaning they do not require optical, electrical or mechanical expertise to setup and operate.
The successful demonstration of remote focusing a multifocal, multiphoton microscope using a spatial light modulator as proposed here will, on further development, provide the medical and biological communities a cost effective means of acquiring volumetric images for fundamental studies in the neurosciences. Neurological studies within living systems pose a particularly challenging scenario, requiring detailed, high-resolution (<1 ?m) images from deep (up to 1 mm) within the highly scattering specimen. The proposed single element detection technique allows unprecedented, quantitative exploration of dynamics within volumes of living tissue, which directly addresses the imaging challenges confounding this community.
