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BIOMEDICAL IMAGING GROUP (BIG)
Laboratoire d'imagerie biomédicale (LIB)
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Design and Validation of a Tool for Neurite Tracing and Analysis in Fluorescence Microscopy Images

E. Meijering, M. Jacob, J.-C.F. Sarria, P. Steiner, H. Hirling, M. Unser

Cytometry Part A, vol. 58A, no. 2, pp. 167-176, April 2004.


Background: For the investigation of the molecular mechanisms involved in neurite outgrowth and differentiation, accurate and reproducible segmentation and quantification of neuronal processes are a prerequisite. To facilitate this task, we developed a semiautomatic neurite tracing technique. This article describes the design and validation of the technique.

Methods: The technique was compared to fully manual delineation. Four observers repeatedly traced selected neurites in 20 fluorescence microscopy images of cells in culture, using both methods. Accuracy and reproducibility were determined by comparing the tracings to high-resolution reference tracings, using two error measures. Labor intensiveness was measured in numbers of mouse clicks required. The significance of the results was determined by a Student t-test and by analysis of variance.

Results: Both methods slightly underestimated the true neurite length, but the differences were not unanimously significant. The average deviation from the true neurite centerline was a factor 2.6 smaller with the developed technique compared to fully manual tracing. Intraobserver variability in the respective measures was reduced by a factor 6.0 and 23.2. Interobserver variability was reduced by a factor 2.4 and 8.8, respectively, and labor intensiveness by a factor 3.3.

Conclusions: Providing similar accuracy in measuring neurite length, significantly improved accuracy in neurite centerline extraction, and significantly improved reproducibility and reduced labor intensiveness, the developed technique may replace fully manual tracing methods.

@ARTICLE(http://bigwww.epfl.ch/publications/meijering0401.html,
AUTHOR="Meijering, E. and Jacob, M. and Sarria, J.-C.F. and Steiner,
	P. and Hirling, H. and Unser, M.",
TITLE="Design and Validation of a Tool for Neurite Tracing and
	Analysis in Fluorescence Microscopy Images",
JOURNAL="Cytometry Part {A}",
YEAR="2004",
volume="58A",
number="2",
pages="167--176",
month="April",
note="")

© 2004 Wiley InterScience®. Personal use of this material is permitted. However, permission to reprint/republish this material for advertising or promotional purposes or for creating new collective works for resale or redistribution to servers or lists, or to reuse any copyrighted component of this work in other works must be obtained from Wiley InterScience®. This material is presented to ensure timely dissemination of scholarly and technical work. Copyright and all rights therein are retained by authors or by other copyright holders. All persons copying this information are expected to adhere to the terms and constraints invoked by each author's copyright. In most cases, these works may not be reposted without the explicit permission of the copyright holder.
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