Polarization contrast optical diffraction tomography.

We demonstrate large scale polarization contrast optical diffraction tomography (ODT). In cross-polarized sample arm detection configuration we determine, from the amplitude of the optical wavefield, a relative measure of the birefringence projection. In parallel-polarized sample arm detection configuration we image the conventional phase projection. For off-axis sample placement we observe for polarization contrast ODT, similar as for phase contrast ODT, a strongly reduced noise contribution. In the limit of small birefringence phase shift δ we demonstrate tomographic reconstruction of polarization contrast images into a full 3D image of an optically cleared zebrafish. The polarization contrast ODT reconstruction shows muscular zebrafish tissue, which cannot be visualized in conventional phase contrast ODT. Polarization contrast ODT images of the zebrafish show a much higher signal to noise ratio (SNR) than the corresponding phase contrast images, SNR=73 and SNR=15, respectively.

Large-scale high-sensitivity optical diffraction tomography of zebrafish.

In this work we demonstrate large-scale high-sensitivity optical diffraction tomography (ODT) of zebrafish. We make this possible by three improvements. First, we obtain a large field of view while still maintaining a high resolution by using a high magnification over numerical aperture ratio digital holography set-up. With the inclusion of phase shifting we operate close to the optimum magnification over numerical aperture ratio. Second, we decrease the noise in the reconstructed images by implementing off-axis sample placement and numerical focus tracking in combination with the acquisition of a large number of projections. Although both techniques lead to an increase in sensitivity independently, we show that combining them is necessary in order to make optimal use of the potential gain offered by each respective method and obtain a refractive index (RI) sensitivity of 8 ⋅ 10 - 5 . Third, we optimize the optical clearing procedure to prevent scattering and refraction to occur. We demonstrate our technique by imaging a zebrafish larva over 13 mm 3 field of view with 4 micrometer resolution. Finally, we demonstrate a clinical application of our technique by imaging an entire adult cryoinjured zebrafish heart.