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Multimodal 3D Imaging of Cells and Tissue, Bridging the Gap Between Clinical and Research Microscopy

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Abstract

Absorption dyes are widely used in traditional cytology and pathology clinical practice, while fluorophores and nanoparticles are more often used in biologic research. Optical projection tomographic microscopy (OPTM) is a platform technology that can image the same specimen in multiple modes in 3D, providing morphologic and molecular information concurrently and in exact co-registration. The depth-of-field of a high numerical aperture objective is extended by scanning the focal plane through the sample to generate an optical projection image. Samples of cells or tissue are brought into the OPTM instrument through a microcapillary tube filled with optical index-matching gel. Multiple optical projection images are taken from different perspectives by rotating the tube. Computed tomography (CT) algorithms are applied to these optical projection images to reconstruct 3D structure of the sample. Image segmentation and analysis based on these 3D images provide quantitative biosignatures for cancer diagnosis that represents a clear improvement over conventional 2D image analysis. In this article, we introduce the OPTM platform, optical Cell-CT, and Tissue-CT instruments, and some applications using these OPTM instruments.

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Acknowledgments

Development of the Cell-CT instrumentation was done in cooperation with VisionGate, Inc., (Phoenix, AZ) at the University of Washington, Eric Seibel (PI). Key technical assistance is provided by the following individuals: Alan Nelson, Mark Fauver, Richard Rahn, Michael Meyer, Thomas Neumann, Jon Hayenga, Christy Lancaster, Wayne Briedford, Anna Touraskaia, David Steinhauer, Tom Abbott, Mathew Watson, Ryland Bryant, Sarah Shimer, Rahul Katdare, Julia Yu, and Greg Kramer. Muntjac cells are a gift from Dr. Roger Schultz and Dr. Lisa McDaniel, Signature Genomic Laboratories, WA. Stellated gold nanoparticles are prepared by Chun-Hsien Wu from Professor Konstantin Sokolov’s lab in the University of Texas in Austin. Cell-CT is a trademark of VisionGate Inc. (www.visiongate3d.com). The Tissue-CT prototype was made possible by an equipment loan of an extended range piezoelectric scanner by Jim Litynski (Piezojena Systems Inc.). Funding is provided by the NSF Collaborative Interdisciplinary Research Program (CBET-1014976, Eric Seibel and Anthony Reeves as PIs). Previous funding includes a gift from VisionGate to Dr. Seibel, and Dr. Seibel and Dr. Reeves have been consultants to VisionGate.

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Authors and Affiliations

  1. Department of Bioengineering, University of Washington, 1705 NE Pacific Street, Seattle, WA, 98195, USA

    Qin Miao

  2. Department of Electrical and Computer Engineering, Cornell University, Ithaca, NY, 14853, USA

    Anthony P. Reeves

  3. VisionGate Inc., Phoenix, AZ, 85004, USA

    Florence W. Patten

  4. Department of Mechanical Engineering, University of Washington, Stevens Way, Seattle, WA, 98195, USA

    Florence W. Patten & Eric J. Seibel

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  1. Qin Miao
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  2. Anthony P. Reeves
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  4. Eric J. Seibel
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Correspondence to Eric J. Seibel.

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Associate Editor Daniel Elson oversaw the review of this article.

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Miao, Q., Reeves, A.P., Patten, F.W. et al. Multimodal 3D Imaging of Cells and Tissue, Bridging the Gap Between Clinical and Research Microscopy. Ann Biomed Eng 40, 263–276 (2012). https://doi.org/10.1007/s10439-011-0411-5

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