Confocal bi-protocol: a new strategy for isotropic 3D live cell imaging

Summary The conventional approach for microscopic 3D cellular imaging is based on axial through-stack image series which has some significant limitations such as anisotropic resolution and axial aberration. To overcome these drawbacks, we have recently introduced an alternative approach based on micro-rotation image series. Unfortunately, this new technique suffers from a huge burden of computation that makes its use quite difficult for current applications. To address these problems we propose a new imaging strategy called bi-protocol, which consists of coupling micro-rotation acquisition and conventional z-stack acquisition. We experimentally prove bi-protocol 3D reconstruction produces similar quality to that of pure micro-rotation, but offers the advantage of reduced computation burden because it uses the z-stack volume to accelerate the registration of the micro-rotation images.