Accurate neuron segmentation method for one-photon calcium imaging videos combining convolutional neural networks and clustering

• Published in: Communications biology Vol. 7; no. 1; pp. 970 - 16
• Main Authors: Bao, Yijun, Gong, Yiyang
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 09-08-2024; Nature Publishing Group; Nature Portfolio
• Subjects: 14; 14/10; 14/35; 14/63; 631/1647/245/2225; 631/378; Algorithms; Animals; Biomedical and Life Sciences; Brain - diagnostic imaging; Brain - metabolism; Calcium - analysis; Calcium - metabolism; Calcium imaging; Cluster Analysis; Image Processing, Computer-Assisted - methods; Information processing; Life Sciences; Mice; Neural networks; Neural Networks, Computer; Neurogenesis; Neuroimaging; Neurons; Neurons - metabolism; Segmentation; Software
• ISSN: 2399-3642; 2399-3642
• DOI: 10.1038/s42003-024-06668-7

One-photon fluorescent calcium imaging helps understand brain functions by recording large-scale neural activities in freely moving animals. Automatic, fast, and accurate active neuron segmentation algorithms are essential to extract and interpret information from these videos. One-photon imaging videos’ low resolution, high noise, and high background fluctuation pose significant challenges. Here, we develop a software pipeline to address the challenges of processing one-photon calcium imaging videos. We extend our previous two-photon active neuron segmentation algorithm, Shallow U-Net Neuron Segmentation (SUNS), to better suppress background fluctuations in one-photon videos. We also develop additional neuron extraction (ANE) to locate small or dim neurons missed by SUNS. To train our segmentation method, we create ground truth neurons by developing a manual labeling pipeline assisted with semi-automatic refinement. Our method is more accurate and faster than state-of-the-art techniques when processing simulated videos and multiple experimental datasets acquired over various brain regions with different imaging conditions.
SUNS2-ANE segments active neurons from one-photon calcium imaging videos accurately and fast. It improves background suppression and extracts additional weak neurons. A separate semi-automatic labeling pipeline generates ground truth for training.