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Journal of Biomedical Optics

Lens-free computational imaging of capillary morphogenesis within three-dimensional substrates
Author(s): John Weidling; Serhan O. Isikman; Alon Greenbaum; Aydogan Ozcan; Elliot L. Botvinick
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Paper 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 (24  mm 2 ) with ∼1.5  μm 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.

Paper Details

Date Published: 13 December 2012
PDF: 7 pages
J. Biomed. Opt. 17(12) 126018 doi: 10.1117/1.JBO.17.12.126018
Published in: Journal of Biomedical Optics Volume 17, Issue 12
Show Author Affiliations
John Weidling, Univ. of California, Irvine (United States)
Serhan O. Isikman, Univ. of California, Los Angeles (United States)
Alon Greenbaum, Univ. of California, Los Angeles (United States)
Aydogan Ozcan, Univ. of California, Los Angeles (United States)
Elliot L. Botvinick, Univ. of California, Irvine (United States)

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