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Journal of Biomedical Optics

High-contrast multilayer imaging of biological organisms through dark-field digital refocusing
Author(s): Ahmad Faridian; Giancarlo Pedrini; Wolfgang Osten
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Paper Abstract

We have developed an imaging system to extract high contrast images from different layers of biological organisms. Utilizing a digital holographic approach, the system works without scanning through layers of the specimen. In dark-field illumination, scattered light has the main contribution in image formation, but in the case of coherent illumination, this creates a strong speckle noise that reduces the image quality. To remove this restriction, the specimen has been illuminated with various speckle-fields and a hologram has been recorded for each speckle-field. Each hologram has been analyzed separately and the corresponding intensity image has been reconstructed. The final image has been derived by averaging over the reconstructed images. A correlation approach has been utilized to determine the number of speckle-fields required to achieve a desired contrast and image quality. The reconstructed intensity images in different object layers are shown for different sea urchin larvae. Two multimedia files are attached to illustrate the process of digital focusing.

Paper Details

Date Published: 12 August 2013
PDF: 7 pages
J. Biomed. Opt. 18(8) 086009 doi: 10.1117/1.JBO.18.8.086009
Published in: Journal of Biomedical Optics Volume 18, Issue 8
Show Author Affiliations
Ahmad Faridian, Institut für Technische Optik (Germany)
Giancarlo Pedrini, Institut für Technische Optik (Germany)
Wolfgang Osten, Institut für Technische Optik (Germany)

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