Abstract
Across basic research studies, cell counting requires significant human time and expertise. Trained experts use thin focal plane scanning to count (click) cells in stained biological tissue.
This computer-assisted process (optical disector) requires a well-trained human to select a unique best z-plane of focus for counting cells of interest. Though accurate, this approach typically requires an hour per case and is prone to inter- and intra-rater errors.
Our group has previously proposed deep learning (DL)-based methods to automate these counts using cell segmentation at high magnification. Here we propose a novel You Only Look Once (YOLO) model that performs cell detection on multi-channel z-plane images (disector stack).
This automated Multiple Input Multiple Output (MIMO) version of the optical disector method uses an entire z-stack of microscopy images as its input, and outputs cell detections (counts) with a bounding box of each cell and class corresponding to the z-plane where the cell appears in best focus. Compared to the previous segmentation methods, the proposed method does not require time- and labor-intensive ground truth segmentation masks for training, while producing comparable accuracy to current segmentation-based automatic counts.
The MIMO-YOLO method was evaluated on systematic-random samples of NeuN-stained tissue sections through the neocortex of mouse brains (n=7). Using a cross validation scheme, this method showed the ability to correctly count total neuron numbers with accuracy close to human experts and with 100% repeatability (Test-Retest).