title: Algorithms for differentiating between images of heterogeneous tissue across fluorescence microscopes.
abstract: Fluorescence microscopy can be used to acquire real-time images of tissue morphology and with appropriate algorithms can rapidly quantify features associated with disease. The objective of this study was to assess the ability of various segmentation algorithms to isolate fluorescent positive features (FPFs) in heterogeneous images and identify an approach that can be used across multiple fluorescence microscopes with minimal tuning between systems. Specifically, we show a variety of image segmentation algorithms applied to images of stained tumor and muscle tissue acquired with 3 different fluorescence microscopes. Results indicate that a technique called maximally stable extremal regions followed by thresholding (MSER + Binary) yielded the greatest contrast in FPF density between tumor and muscle images across multiple microscopy systems.
authors.affiliation: Department of Biomedical Engineering, Duke University, Durham, North Carolina, USA; chitalia.rhea@gmail.com.
authors.forename: Rhea
authors.initials: R
authors.lastname: Chitalia
authors.affiliation: Department of Biomedical Engineering, Duke University, Durham, North Carolina, USA; jenna.mueller@duke.edu.
authors.forename: Jenna
authors.initials: J
authors.lastname: Mueller
authors.affiliation: Department of Biomedical Engineering, Duke University, Durham, North Carolina, USA.
authors.forename: Henry L
authors.initials: HL
authors.lastname: Fu
authors.affiliation: Department of Pharmacology & Cancer Biology, Duke University School of Medicine, Durham, North Carolina, USA.
authors.forename: Melodi Javid
authors.initials: MJ
authors.lastname: Whitley
authors.affiliation: Department of Pharmacology & Cancer Biology, Duke University School of Medicine, Durham, North Carolina, USA; Department of Radiation Oncology, Duke University School of Medicine, Durham, North Carolina, USA.
authors.forename: David G
authors.initials: DG
authors.lastname: Kirsch
authors.affiliation: Department of Biomedical Engineering, Tulane University, New Orleans, Louisiana, USA.
authors.forename: J Quincy
authors.initials: JQ
authors.lastname: Brown
authors.affiliation: Department of Electrical and Computer Engineering, University of Wisconsin-Madison, Madison, Wisconsin, USA.
authors.forename: Rebecca
authors.initials: R
authors.lastname: Willett
authors.affiliation: Department of Biomedical Engineering, Duke University, Durham, North Carolina, USA.
authors.forename: Nimmi
authors.initials: N
authors.lastname: Ramanujam
keywords: (100.0100) Image processing; (180.0180) Microscopy