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Plant Physiol, May 2000, Vol. 123, pp. 3-16

Optical Coherence Microscopy. A Technology for Rapid, in Vivo, Non-Destructive Visualization of Plants and Plant Cells1,[w]

James W. Hettinger, Matthew de la Peña Mattozzi, Whittier R. Myers,2 Mary E. Williams, Aaron Reeves, Ronald L. Parsons, Richard C. Haskell, Daniel C. Petersen, Ruye Wang, and June I. Medford*

Department of Biology, Colorado State University, Fort Collins, Colorado 80523-1878 (J.W.H., A.R., R.L.P., J.I.M.); and Departments of Biology (M.d.l.P.M., M.E.W.), Physics (W.R.M., R.C.H., D.C.P.), and Engineering (R.W.), Harvey Mudd College, Claremont, California 91711

We describe the development and utilization of a new imaging technology for plant biology, optical coherence microscopy (OCM), which allows true in vivo visualization of plants and plant cells. This novel technology allows the direct, in situ (e.g. plants in soil), three-dimensional visualization of cells and events in shoot tissues without causing damage. With OCM we can image cells or groups of cells that are up to 1 mm deep in living tissues, resolving structures less than 5 µm in size, with a typical collection time of 5 to 6 min. OCM measures the inherent light-scattering properties of biological tissues and cells. These optical properties vary and provide endogenous developmental markers. Singly scattered photons from small (e.g. 5 × 5 × 10 µm) volume elements (voxels) are collected, assembled, and quantitatively false-colored to form a three-dimensional image. These images can be cropped or sliced in any plane. Adjusting the colors and opacities assigned to voxels allows us to enhance different features within the tissues and cells. We show that light-scattering properties are the greatest in regions of the Arabidopsis shoot undergoing developmental processes. In large cells, high light scattering is produced from nuclei, intermediate light scatter is produced from cytoplasm, and little if any light scattering originates from the vacuole and cell wall. OCM allows the rapid, repetitive, non-destructive collection of quantitative data about inherent properties of cells, so it provides a means of continuously monitoring plants and plant cells during development and in response to exogenous stimuli.

1 This work was supported by the National Science Foundation (grant no. DBI-9612240 to R.C.H., D.C.P., R.W., M.E.W., and Scott Fraser [California Institute of Technology]).

2 Present address: Department of Physics, 366 LeConte Hall, University of California, Berkeley, CA 94720-7300.

[w]  The on-line version of this article contains Web-only data. This version is available at www.plantphysiol.org.

* Corresponding author; e-mail medford{at}lamar.colostate.edu; fax 970-491-0649.

© 2000 American Society of Plant Physiologists


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