Abstract
We report the experimental implementation of optical diffraction tomography for quantitative 3D mapping of refractive index in live biological cells. Using a heterodyne Mach-Zehnder interferometer, we record complex field images of light transmitted through a sample with varying directions of illumination. To quantitatively reconstruct the 3D map of complex refractive index in live cells, we apply optical diffraction tomography based on the Rytov approximation. In this way, the effect of diffraction is taken into account in the reconstruction process and diffraction-free high resolution 3D images are obtained throughout the entire sample volume. The quantitative refractive index map can potentially serve as an intrinsic assay to provide the molecular concentrations without the addition of exogenous agents and also to provide a method for studying the light scattering properties of single cells.
Highlights
Refractive index serves as an important intrinsic contrast agent in visualizing nearly transparent living biological cells
We have developed tomographic phase microscopy (TPM) [16] for quantitative 3D mapping of refractive index in live cells in their native state
We report the first experimental implementation of optical diffraction tomography to image live biological cells and provide quantitative 3D refractive index maps
Summary
Refractive index serves as an important intrinsic contrast agent in visualizing nearly transparent living biological cells. Several experimental studies have implemented diffraction tomography in the optical regime [13, 15, 21], but one [13] imaged only non-biological samples and the others [15, 21] did not provide calibrated values for refractive index. We report the first experimental implementation of optical diffraction tomography to image live biological cells and provide quantitative 3D refractive index maps. By employing optical diffraction tomography based on the Rytov approximation, we take diffraction into account and produce high resolution 3D refractive index maps for the entire cell without need for propagation in the reconstruction algorithm. They provide a means of studying the light scattering of single cells [24], which may lead to develop in-vivo light scattering instruments for disease diagnosis
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