Neurosphere segmentation in brightfield images
Jierong Cheng, Wei Xiong, Shue Ching Chia, Joo‐Hwee Lim, Shvetha Sankaran, Sohail Ahmed · Proceedings of SPIE, the International Society for Optical Engineering/Proceedings of SPIE · 2014
The challenge of segmenting neurospheres (NSPs) from brightfield images includes uneven background illumination (vignetting), low contrast and shadow-casting appearance near the well wall. We propose a pipeline for neurosphere segmentation in brightfield images, focusing on shadow-casting removal. Firstly, we remove vignetting by creating a synthetic blank field image from a set of brightfield images of the whole well. Then, radial line integration is proposed to remove the shadow-casting and therefore facilitate automatic segmentation. Furthermore, a weighted bi-directional decay function is introduced to prevent undesired gradient effect of line integration on NSPs without shadow-casting. Afterward, multiscale Laplacian of Gaussian (LoG) and localized region-based level set are used to detect the NSP boundaries. Experimental results show that our proposed radial line integration method (RLI) achieves higher detection accuracy over existing methods in terms of precision, recall and F-score with less computational time.