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Research Papers: Imaging

Lens-free computational imaging of capillary morphogenesis within three-dimensional substrates

[+] Author Affiliations
John Weidling

University of California Irvine, Biomedical Engineering Department, Irvine, California

Serhan O. Isikman, Alon Greenbaum

University of California Los Angeles, Electrical Engineering Department, Los Angeles, California

Aydogan Ozcan

University of California Los Angeles, Electrical Engineering Department, Los Angeles, California

University of California Los Angeles, Bioengineering Department, Los Angeles, California

University of California Los Angeles, California NanoSystems Institute, Los Angeles, California

Elliot Botvinick

University of California Irvine, Biomedical Engineering Department, Irvine, California

University of California Irvine, Beckman Laser Institute, Irvine, California

University of California Irvine, Edwards Lifesciences Center for Advanced Cardiovascular Technology, Irvine, California

J. Biomed. Opt. 17(12), 126018 (Dec 13, 2012). doi:10.1117/1.JBO.17.12.126018
History: Received August 19, 2012; Revised November 14, 2012; Accepted November 19, 2012
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Abstract.  Endothelial cells cultured in three-dimensional (3-D) extracellular matrices spontaneously form microvessels in response to soluble and matrix-bound factors. Such cultures are common for the study of angiogenesis and may find widespread use in drug discovery. Vascular networks are imaged over weeks to measure the distribution of vessel morphogenic parameters. Measurements require micron-scale spatial resolution, which for light microscopy comes at the cost of limited field-of-view (FOV) and shallow depth-of-focus (DOF). Small FOVs and DOFs necessitate lateral and axial mechanical scanning, thus limiting imaging throughput. We present a lens-free holographic on-chip microscopy technique to rapidly image microvessels within a Petri dish over a large volume without any mechanical scanning. This on-chip method uses partially coherent illumination and a CMOS sensor to record in-line holographic images of the sample. For digital reconstruction of the measured holograms, we implement a multiheight phase recovery method to obtain phase images of capillary morphogenesis over a large FOV (24mm2) with 1.5μm spatial resolution. On average, measured capillary length in our method was within approximately 2% of lengths measured using a 10× microscope objective. These results suggest lens-free on-chip imaging is a useful toolset for high-throughput monitoring and quantitative analysis of microvascular 3-D networks.

Figures in this Article
© 2012 Society of Photo-Optical Instrumentation Engineers

Citation

John Weidling ; Serhan O. Isikman ; Alon Greenbaum ; Aydogan Ozcan and Elliot Botvinick
"Lens-free computational imaging of capillary morphogenesis within three-dimensional substrates", J. Biomed. Opt. 17(12), 126018 (Dec 13, 2012). ; http://dx.doi.org/10.1117/1.JBO.17.12.126018


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