Constrained watershed method to infer morphology of mammalian cells in microscopic images

Precise information about the size, shape, temporal dynamics, and spatial distribution of cells is beneficial for the understanding of cell behavior and may play a key role in drug development, regenerative medicine, and disease research. The traditional method of manual observation and measurement of cells from microscopic images is tedious, expensive, and time consuming. Thus, automated methods are in high demand, especially given the increasing quantity of cell data being collected. In this article, an automated method to measure cell morphology from microscopic images is proposed to outline the boundaries of individual hematopoietic stem cells (HSCs). The proposed method outlines the cell regions using a constrained watershed method which is derived as an inverse problem. The experimental results generated by applying the proposed method to different HSC image sequences showed robust performance to detect and segment individual and dividing cells. The performance of the proposed method for individual cell segmentation for single frame high‐resolution images was more than 97%, and decreased slightly to 90% for low‐resolution multiframe stitched images. © 2010 International Society for Advancement of Cytometry

[1]  Paul Fieguth,et al.  Watershed deconvolution for cell segmentation , 2008, 2008 30th Annual International Conference of the IEEE Engineering in Medicine and Biology Society.

[2]  P. Fieguth,et al.  Stem-Cell Localization: A Deconvolution Problem , 2007, 2007 29th Annual International Conference of the IEEE Engineering in Medicine and Biology Society.

[3]  Matthias Gunzer,et al.  Altered cellular dynamics and endosteal location of aged early hematopoietic progenitor cells revealed by time-lapse intravital imaging in long bones. , 2009, Blood.

[4]  H. Müller-Hermelink,et al.  Image analysis detects lineage-specific morphologic markers in leukemic blast cells. , 1996, American journal of clinical pathology.

[5]  Arnold W. M. Smeulders,et al.  Segmentation of cell clusters by nearest neighbour graphs , 1997 .

[6]  Dorin Comaniciu,et al.  Shape-based image indexing and retrieval for diagnostic pathology , 1998, Proceedings. Fourteenth International Conference on Pattern Recognition (Cat. No.98EX170).

[7]  Eric Jervis,et al.  High-resolution video monitoring of hematopoietic stem cells cultured in single-cell arrays identifies new features of self-renewal. , 2006, Proceedings of the National Academy of Sciences of the United States of America.

[8]  Paul W. Fieguth,et al.  Extended-Hungarian-JPDA: Exact Single-Frame Stem Cell Tracking , 2007, IEEE Transactions on Biomedical Engineering.

[9]  Serge Beucher,et al.  Use of watersheds in contour detection , 1979 .

[10]  N.N. Kachouie,et al.  A Statistical Thresholding Method for Cell Tracking , 2006, 2006 IEEE International Symposium on Signal Processing and Information Technology.

[11]  John F. Canny,et al.  A Computational Approach to Edge Detection , 1986, IEEE Transactions on Pattern Analysis and Machine Intelligence.

[12]  Eric Jervis,et al.  Design and analysis of a long‐term live‐cell imaging chamber for tracking cellular dynamics within cultured human islets of Langerhans , 2007, Biotechnology and bioengineering.

[13]  M. Collector,et al.  Hematopoietic stem cell tracking in vivo: a comparison of short-term and long-term repopulating cells. , 1999, Blood.

[14]  Dorin Comaniciu,et al.  Cell image segmentation for diagnostic pathology , 2001 .

[15]  Tomasz Markiewicz,et al.  Myelogenous Leukemia Cell Image Preprocessing for Feature Generation , .

[16]  D. Kent,et al.  Identification of a new intrinsically timed developmental checkpoint that reprograms key hematopoietic stem cell properties , 2007, Proceedings of the National Academy of Sciences.

[17]  Josef Kittler,et al.  Minimum error thresholding , 1986, Pattern Recognit..

[18]  B. Palsson,et al.  Murine Sca-1(+)/Lin(-) cells and human KG1a cells exhibit multiple pseudopod morphologies during migration. , 2002, Experimental hematology.

[19]  N. Otsu.  A threshold selection method from gray level histograms , 1979 .

[20]  P. Wernet,et al.  Segregation of lipid raft markers including CD133 in polarized human hematopoietic stem and progenitor cells. , 2004, Blood.

[21]  Kenong Wu,et al.  Live cell image segmentation , 1995, IEEE Transactions on Biomedical Engineering.

[22]  Pierre Machart.  Morphological Segmentation , 2009 .

[23]  Paul W. Fieguth,et al.  Probabilistic Model-Based Cell Tracking , 2006, Int. J. Biomed. Imaging.