This dataset contains 3D confocal microscope images of PC12 cells after various times of treatment of Nerve Growth Factor and labeling with Mito Red. It is from: "Xiongtao Ruan, Gregory R. Johnson, Iris Bierschenk, Roland Nitschke, Melanie Boerries, Hauke Busch and Robert F. Murphy, Image-derived Models of Cell Organization Changes During Differentiation of PC12 Cells" (submitted to PLOS Computational Biology) The technical details for cell culture and imaging are as follows (copied from the above manuscirpt): Cell Culture and Experimental conditions PC12 cells (between 6 and 10 passages) were obtained from ATCC (American Type Culture Collection, UK) and were cultured in RPMi medium containing 10% horse serum (HS), 5% fetal calf serum (FCS) 1% L-Glutamine and Penicillin/Streptomycin at 37°C in 5% CO 2 . Cells were plated on collagen coated 35mm glass-bottom ibiTreat dishes and were allowed to adhere for 24 hours. Two types of experiments were performed. Cells were either treated with 50ng/ml rat Nerve Growth Factor (NG; Promega, Madison, WI, USA) at 0, 12, 24, 36 and 48h prior to imaging at the same time, or were treated at the same time and imaged at 24 h increments up to 96 h after treatment. An hour prior to imaging, cells were stained at 37° C with a 0.5uM solution of Mito Red (Sigma-Aldrich, Munich, Germany) for 5 minutes, rinsed with PBS and placed in 1ml of growth media without Phenol red. Microscopy Cells were imaged on an Axio Observer.Z1 (Carl Zeiss Microscopy, Jena, Germany) microscope equipped with a spinning disk (CSU-22, Yokogawa, Japan) with an EX-Plan-Neofluar 40x/1.30 Oil objective. The sample voxel size was 0.161 um x 0.161 um x 0.340 um and 59 slices were taken with a 150ms exposure time at 12-bit pixel depth. Imaged cells were manually selected to not be in contact with other cells. Due to the sensitivity of cells to phototoxicity, approximately 10 fields were imaged per plate. Between 172 and 98 cells were imaged at each time point for the 48h experiment and between 46 and 89 cells per time point in the 96h experiment. Organization and formats of the datasets The dataset consists of images from two experiments with directory names "exp48hr" and "exp96hr" (please see the manuscript for a description of the experiments). The images are contained in 3 .tgz files created by the Linux "tar" command, e.g., "tar czf PC12_images_exp96.tgz images/exp96hr" The three files are PC12_images_exp48A.tgz (4.78 GB) PC12_images_exp48B.tgz (7.93 GB) PC12_images_exp96.tgz (6.15 GB) Within each experiment directory, there are directories for different time points and dates when. Each directory name is in the format "Xhr-M-D_N-Lms" where X represents the time after NGF addition, M and D are the month and date of acquisition (in 2013), N is the number of the slide, and L is the exposure time. Each directory contains approximately 50 (96 hr experiment) or 100 (48 hr experiment) OME-TIFF images of different fields from a given time of treatment. Each OME-TIFF file contains a 3D z-stack with only one fluorescent channel. There are a total of 616 files, with 414 files for the 48hr experiment and 202 for the 96hr experiment.