Research Papers

Dual-modal three-dimensional imaging of single cells with isometric high resolution using an optical projection tomography microscope

[+] Author Affiliations
Qin Miao

University of Washington, Department of Bioengineering, 204 Fluke Hall, Seattle, Washington 98195

J. Richard Rahn, Anna Tourovskaia, Michael G. Meyer, Thomas Neumann, Alan C. Nelson

Vision Gate Inc., 1509 56th Avenue Court North West, Gig Harbor, Washington 98335

Eric J. Seibel

University of Washington, Dept. of Mechanical Engineering, P.O. Box 352600, Seattle, Washington 98195

J. Biomed. Opt. 14(6), 064035 (December 21, 2009). doi:10.1117/1.3275470
History: Received April 15, 2009; Revised October 15, 2009; Accepted October 15, 2009; Published December 21, 2009; Online December 21, 2009
Text Size: A A A

The practice of clinical cytology relies on bright-field microscopy using absorption dyes like hematoxylin and eosin in the transmission mode, while the practice of research microscopy relies on fluorescence microscopy in the epi-illumination mode. The optical projection tomography microscope is an optical microscope that can generate 3-D images of single cells with isometric high resolution both in absorption and fluorescence mode. Although the depth of field of the microscope objective is in the submicron range, it can be extended by scanning the objective’s focal plane. The extended depth of field image is similar to a projection in a conventional x-ray computed tomography. Cells suspended in optical gel flow through a custom-designed microcapillary. Multiple pseudoprojection images are taken by rotating the microcapillary. After these pseudoprojection images are further aligned, computed tomography methods are applied to create 3-D reconstruction. 3-D reconstructed images of single cells are shown in both absorption and fluorescence mode. Fluorescence spatial resolution is measured at 0.35μm in both axial and lateral dimensions. Since fluorescence and absorption images are taken in two different rotations, mechanical error may cause misalignment of 3-D images. This mechanical error is estimated to be within the resolution of the system.

Figures in this Article
© 2009 Society of Photo-Optical Instrumentation Engineers

Citation

Qin Miao ; J. Richard Rahn ; Anna Tourovskaia ; Michael G. Meyer ; Thomas Neumann, et al.
"Dual-modal three-dimensional imaging of single cells with isometric high resolution using an optical projection tomography microscope", J. Biomed. Opt. 14(6), 064035 (December 21, 2009). ; http://dx.doi.org/10.1117/1.3275470


Tables

Access This Article
Sign in or Create a personal account to Buy this article ($20 for members, $25 for non-members).

Some tools below are only available to our subscribers or users with an online account.

Related Content

Customize your page view by dragging & repositioning the boxes below.

Related Book Chapters

Topic Collections

Advertisement
  • Don't have an account?
  • Subscribe to the SPIE Digital Library
  • Create a FREE account to sign up for Digital Library content alerts and gain access to institutional subscriptions remotely.
Access This Article
Sign in or Create a personal account to Buy this article ($20 for members, $25 for non-members).
Access This Proceeding
Sign in or Create a personal account to Buy this article ($15 for members, $18 for non-members).
Access This Chapter

Access to SPIE eBooks is limited to subscribing institutions and is not available as part of a personal subscription. Print or electronic versions of individual SPIE books may be purchased via SPIE.org.