Multiview deconvolution two-photon microscopy with orthogonal objectives

Multiview deconvolution microscopy achieves near isotropic resolution by combining volumetric images captured from multiple angles. We introduce a novel two-photon fluorescence microscope with two orthogonal objectives scanning the same focal line. Each image strip is rapidly acquired from both directions, minimizing misalignment and delays between views. Using non-descanned silicon photomultipliers (SiPM), our system supports multiple simultaneous fluorescence channels. We demonstrate enhanced axial resolution and high-throughput volumetric imaging of dermatologic biopsy samples, highlighting this method’s potential in biomedical imaging research.