Title: Towards fully automated identification of vesicle-membrane fusion events in TIRF microscopy

Authors: Katarina Mele, Pascal Vallotton, James G. Burchfield, David E. James, William E. Hughes

Addresses: Biotech Imaging, CSIRO Mathematical and Information Sciences, Locked Bag 17, North Ryde, NSW 1670, Australia; Diabetes and Obesity Research Program, The Garvan Institute of Medical Research, 384 Victoria Street, Sydney, NSW, 2010, Australia. ' Biotech Imaging, CSIRO Mathematical and Information Sciences, Locked Bag 17, North Ryde, NSW 1670, Australia. ' Diabetes and Obesity Research Program, The Garvan Institute of Medical Research, 384 Victoria Street, Sydney, NSW, 2010, Australia. ' Diabetes and Obesity Research Program, The Garvan Institute of Medical Research, 384 Victoria Street, Sydney, NSW, 2010, Australia. ' Diabetes and Obesity Research Program, The Garvan Institute of Medical Research, 384 Victoria Street, Sydney, NSW, 2010, Australia

Abstract: Total internal reflection fluorescence microscopy (TIRF-M) is imposing itself as the tool of choice for studying biological activity in close proximity to the plasma membrane. For example, the exquisite selectivity of TIRF-M allows monitoring the movement of GFP-tagged vesicles and their recruitment to the plasma membrane of cells. We present a novel computer vision system for automatically identifying elusive vesicle-plasma membrane fusion events. Our method is based on robust object tracking and matched filtering. It should accelerate the quantification of TIRF-M data and allow the extraction of more biological information from image data to support research in diabetes and obesity.

Keywords: vesicles; vesicle tracking; vesicle fusion; eGFP-phogrin; GFP-GLUT4; IRAP-pHluorin; total internal reflection fluorescence microscopy; TIRF-M; plasma membrane; computer vision; object tracking; matched filtering; diabetes research; obesity research.

DOI: 10.1504/IJCAET.2009.028555

International Journal of Computer Aided Engineering and Technology, 2009 Vol.1 No.4, pp.502 - 515

Available online: 18 Sep 2009 *

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