Suite3D: Volumetric cell detection for two-photon microscopy

In two-photon imaging of neuronal activity it is common to acquire 3-dimensional volumes. However, these volumes are typically processed plane by plane, leading to duplicated cells across planes, reduced signal-to-noise ratio per cell, uncorrected axial movement, and missed cells. To overcome these limitations, we introduce Suite3D, a volumetric cell detection pipeline. Suite3D corrects for 3D brain motion, estimating axial motion and improving estimates of lateral motion. It detects neurons using 3D correlation, which improves the signal-to-background ratio and detectability of cells. Finally, it performs 3D segmentation, detecting cells across imaging planes. We validated Suite3D with data from conventional multi-plane microscopes and advanced volumetric microscopes, at various resolutions and in various brain regions. Suite3D successfully detected cells appearing on multiple imaging planes, improving cell detectability and signal quality, avoiding duplications, and running >20x faster than a prior volumetric pipeline. Suite3D offers a powerful solution for analyzing volumetric two-photon data.