A major challenge in the life sciences is to gain access across scales, from the molecular building blocks, via cells, to organization and dynamics at the composite tissue level. Fluorescence microscopy as the primary biological imaging tool has partly addressed these needs with three major revolutions. The first is the use of 2D sheets of light for illumination so as to only illuminate the part of the sample volume that is image on the camera, which minimizes light induced damage to the cells. The second is a suite of super-resolution techniques where single molecule imaging and patterned illumination are used to image down to and below the 100 nm scale. The third is the use of adaptive optical techniques for imaging heterogeneous samples such as cells or tissues, or for manipulating the illumination and/or imaging light field to a desired shape.
The next frontier in microscopy lies in overcoming trade-offs between imaged volume, resolution, and sample longevity by combining and extending these three innovations. The goal of the project is to create a microscopy platform in which a sequence of patterned light sheets is applied to the sample, where the 2D light sheet pattern, as well as the position and orientation of the 2D light sheet in 3D space is changed in the sequence. This creates a very large diversity of 3D illumination patterns, and this is what enables isotropic super-resolution across large volumes. A second crucial ingredient is the use of computational reconstruction techniques to render the final volumetric image. Taking the heterogeneity of the imaging conditions across the cellular or tissue volume into account is the major goal of algorithm development.
