Gold finder: a computer method for fast automatic double gold labeling detection, counting, and color overlay in electron microscopic images

This work presents a computerized method to identify, detect, evaluate, and, by colored overlay, present gold particle pairs in electron microscopy (EM), even in wide-field views. Double gold immunolabeled specimens were analyzed in a LEO 912 electron microscope equipped with a 2k×2k-pixel slow-scan...

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Bibliographic Details
Published in:Journal of structural biology 2003-03, Vol.141 (3), p.228-239
Main Authors: Monteiro-Leal, Luiz Henrique, Troster, Helmut, Campanati, Loraine, Spring, Herbert, F. Trendelenburg, Michael
Format: Article
Language:English
Subjects:
Animals
Automated detection
Electron microscopy
Giardia lamblia - ultrastructure
Gold
Gold particles
Image Processing, Computer-Assisted
Immunohistochemistry - methods
Immunolabeling
Microscopy, Electron - instrumentation
Microscopy, Electron - methods
Microscopy, Video - methods
Slow-scan cooled CCD camera
Software
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Summary:This work presents a computerized method to identify, detect, evaluate, and, by colored overlay, present gold particle pairs in electron microscopy (EM), even in wide-field views. Double gold immunolabeled specimens were analyzed in a LEO 912 electron microscope equipped with a 2k×2k-pixel slow-scan cooled CCD camera connected to a computer with analySIS 3.1 PRO image processing software. The acquisition of a high-resolution and high-dynamic-range image by the camera allowed correct segmentation of the gold particles, separating them from other cell structures and from the substrate. Particle identification was performed by a classification module designed by us. Based on shape and size, the computer recognized the group of small particles and classified them as either singular or clustered and differentiated these from the single bigger type. The final image shows the particle types separated and colored, and indicates the total number of objects encountered in the specific region of interest. Moreover, a montage tool allowed us to obtain final representative images of large microscopic fields, which on analysis by the Gold Finder module provided information on the distribution and localization of antigens comparable to that provided by the wide-field light microscope images.
ISSN:1047-8477
1095-8657
DOI:10.1016/S1047-8477(02)00624-X