Maximum intensity projection of a dendrite in organotypic hippocampal slice culture labeled by cytosolic eGFP expression (Thy1-eGFP mouse line, confocal) (top left). The positive label served as ground truth for spine detection, with individual spines numbered. Calibration bar represents raw photon counts. Plane from isotropically super-resolved, volumetric LIONESS acquisition used as source data for segmentation with arrow indicating the same dendrite in the extracellularly labeled tissue (maximum intensity projection spanning 150 nm) (bottom left). Fully manual spine detection from LIONESS imaging data by a segmenter blinded to the eGFP channel (BS, blinded segmenter) (top right). Examples of missed and false-positive spines are indicated by white and yellow arrows, respectively. Spine detection in 3D reconstruction after automated segmentation and proofreading of LIONESS imaging data by the experimenter who recorded the data (automated plus proofreading, A + P) (bottom right). As this person was not blinded to the eGFP channel, this indicates which spines can be retrieved from LIONESS but does not serve as an independent control.
