title: Evaluation of methods for detection of fluorescence labeled subcellular objects in microscope images.
abstract: BACKGROUND
  Several algorithms have been proposed for detecting fluorescently labeled subcellular objects in microscope images. Many of these algorithms have been designed for specific tasks and validated with limited image data. But despite the potential of using extensive comparisons between algorithms to provide useful information to guide method selection and thus more accurate results, relatively few studies have been performed.
  
  
  RESULTS
  To better understand algorithm performance under different conditions, we have carried out a comparative study including eleven spot detection or segmentation algorithms from various application fields. We used microscope images from well plate experiments with a human osteosarcoma cell line and frames from image stacks of yeast cells in different focal planes. These experimentally derived images permit a comparison of method performance in realistic situations where the number of objects varies within image set. We also used simulated microscope images in order to compare the methods and validate them against a ground truth reference result. Our study finds major differences in the performance of different algorithms, in terms of both object counts and segmentation accuracies.
  
  
  CONCLUSIONS
  These results suggest that the selection of detection algorithms for image based screens should be done carefully and take into account different conditions, such as the possibility of acquiring empty images or images with very few spots. Our inclusion of methods that have not been used before in this context broadens the set of available detection methods and compares them against the current state-of-the-art methods for subcellular particle detection.
authors.affiliation: Department of Signal Processing, Tampere University of Technology, Tampere, 33101, Finland. pekka.ruusuvuori@tut.fi
authors.forename: Pekka
authors.initials: P
authors.lastname: Ruusuvuori
authors.forename: Tarmo
authors.initials: T
authors.lastname: Aijö
authors.forename: Sharif
authors.initials: S
authors.lastname: Chowdhury
authors.forename: Cecilia
authors.initials: C
authors.lastname: Garmendia-Torres
authors.forename: Jyrki
authors.initials: J
authors.lastname: Selinummi
authors.forename: Mirko
authors.initials: M
authors.lastname: Birbaumer
authors.forename: Aimée M
authors.initials: AM
authors.lastname: Dudley
authors.forename: Lucas
authors.initials: L
authors.lastname: Pelkmans
authors.forename: Olli
authors.initials: O
authors.lastname: Yli-Harja
mesh_terms: D000465:Algorithms; D045744:Cell Line, Tumor; D002478:Cells, Cultured / Q000648:ultrastructure; D006801:Humans; D007090:Image Interpretation, Computer-Assisted / Q000379:methods*; D008856:Microscopy, Fluorescence / Q000379:methods*; D012680:Sensitivity and Specificity
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