Fluorescence microscopy allows researchers to study the structure and function of the brain in both fixed samples and in vivo. Laser scanning microscopy, confocal and multi-photon, serve as the standard imaging approaches for imaging into scattering samples. However, due to the light scattering properties of brain tissue, LSM suffers in both depth penetration and resolution. By combining the unique Airyscan detection concept with multiphoton excitation, the resolution, signal-to-noise, and speed benefits of Airyscan can be extended to deeper layers of the cortex (2-3x deeper than traditional confocal). In addition, combining Airyscan with GRIN lens technology enables increased resolution and signal-to-noise while imaging regions of the brain that are unreachable with traditional in vivo microscopy.
To add additional context to the data, Airyscan imaging through a GRIN lens can be correlated with freely behaving imaging. Miniature microscopes such as the Inscopix nVista™ and nVoke™ systems can be used for cellular-resolution imaging and optogenetic manipulation in freely behaving animals to study functional network activity. Methodology has been developed to register the exact same neurons imaged with the Inscopix miniature microscopes to neurons imaged with Airyscan. Hardware and analysis tools have been developed that enable recording from the same focal plane between the two modalities and correct for the different scale, rotation, and elastic deformations between the images. The ability to register data from freely behaving imaging experiments to high resolution Airyscan images provides crucial links between activity dynamics and anatomical, molecular, and/or connectivity profiles of distinct neuronal populations and can enable exciting new insights into brain health and disease.