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Optics Express

Optics Express

  • Editor: Michael Duncan
  • Vol. 13, Iss. 11 — May. 30, 2005
  • pp: 4210–4223

Three-dimensional imaging of single isolated cell nuclei using optical projection tomography

Mark Fauver, Eric J. Seibel, J. Richard Rahn, Michael G. Meyer, Florence W. Patten, Thomas Neumann, and Alan C. Nelson  »View Author Affiliations

Optics Express, Vol. 13, Issue 11, pp. 4210-4223 (2005)

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A method is presented for imaging single isolated cell nuclei in 3D, employing computed tomographic image reconstruction. The system uses a scanning objective lens to create an extended depth-of-field (DOF) image similar to a projection or shadowgram. A microfabricated inverted v-groove allows a microcapillary tube to be rotated with sub-micron precision, and refractive index matching within 0.02 both inside and outside the tube keeps optical distortion low. Cells or bare cell nuclei are injected into the tube and imaged in 250 angular increments from 0 to 180 degrees to collect 250 extended DOF images. After these images are further aligned, the filtered backprojection algorithm is applied to compute the 3D image. To estimate the cutoff spatial frequency in the projection image, a spatial frequency ratio function is calculated by comparing the extended depth-of-field image of a typical cell nucleus to the fixed focus image. To assess loss of resolution from fixed focus image to extended DOF image to 3D reconstructed image, the 10–90% rise distance is measured for a dyed microsphere. The resolution is found to be 0.9 µm for both extended DOF images and 3D reconstructed images. Surface and translucent volume renderings and cross-sectional slices of the 3D images are shown of a stained nucleus from fibroblast and cancer cell cultures with added color histogram mapping to highlight 3D chromatin structure.

© 2005 Optical Society of America

OCIS Codes
(110.6880) Imaging systems : Three-dimensional image acquisition
(110.6960) Imaging systems : Tomography
(170.0170) Medical optics and biotechnology : Medical optics and biotechnology
(170.1530) Medical optics and biotechnology : Cell analysis
(170.3880) Medical optics and biotechnology : Medical and biological imaging
(180.0180) Microscopy : Microscopy
(180.6900) Microscopy : Three-dimensional microscopy

ToC Category:
Research Papers

Original Manuscript: April 1, 2005
Revised Manuscript: May 17, 2005
Published: May 30, 2005

Mark Fauver, Eric Seibel, J. Richard Rahn, Michael Meyer, Florence Patten, Thomas Neumann, and Alan Nelson, "Three-dimensional imaging of single isolated cell nuclei using optical projection tomography," Opt. Express 13, 4210-4223 (2005)

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