Label-free three-dimensional reconstruction of biological samples
(Conference Presentation)

Sherazade Aknoun; Pierre Bon; Julien Savatier; Serge Monneret; Benoit F. Wattellier

doi:10.1117/12.2212943

27 April 2016 Label-free three-dimensional reconstruction of biological samples (Conference Presentation)

Sherazade Aknoun, Pierre Bon, Julien Savatier, Serge Monneret, Benoit F. Wattellier

Proceedings Volume 9713, Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XXIII; 97131L (2016) https://doi.org/10.1117/12.2212943
Event: SPIE BiOS, 2016, San Francisco, California, United States

Abstract

We describe the use of spatially incoherent illumination combined with quantitative phase imaging (QPI) [1] to make tridimensional reconstruction of semi-transparent biological samples. Quantitative phase imaging is commonly used with coherent illumination for the relatively simple interpretation of the phase measurement. We propose to use spatially incoherent illumination which is known to increase lateral and axial resolution compared to classical coherent illumination. The goal is to image thick samples with intracellular resolution [2]. The 3D volume is imaged by axially scanning the sample with a quadri-wave lateral shearing interferometer used as a conventional camera while using spatially incoherent white-light illumination (native microscope halogen source) or NIR light. We use a non-modified inverted microscope equipped with a Z-axis piezo stage. A z-stack is recorded by objective translation along the optical axis. The main advantages of this approach are its easy implementation, compared to the other state-of-the-art diffraction tomographic setups, and its speed which makes even label-free 3D living sample imaging possible. A deconvolution algorithm is used to compensate for the loss in contrast due to spatially incoherent illumination. This makes the tomographic volume phase values quantitative. Hence refractive index could be recovered from the optical slices. We will present tomographic reconstruction of cells, thick fixed tissue of few tens of micrometers using white light, and the use of NIR light to reach deeper planes in the tissue.

Conference Presentation

Citation Download Citation

Sherazade Aknoun, Pierre Bon, Julien Savatier, Serge Monneret, and Benoit F. Wattellier "Label-free three-dimensional reconstruction of biological samples (Conference Presentation)", Proc. SPIE 9713, Three-Dimensional and Multidimensional Microscopy: Image Acquisition and Processing XXIII, 97131L (27 April 2016); https://doi.org/10.1117/12.2212943

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KEYWORDS

3D image processing

Tissue optics

Tomography

Microscopes

Near infrared

Phase imaging

3D metrology

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