In this paper, we demonstrate the decoupling of complex refractive index in transport of intensity diffraction tomography (TIDT) microscopy under partially coherent illumination. Specifically, with a closed-form Tikhonov-regularized solution, this method enables the decoupling of the RI and absorption from the two through-focus intensity stacks. We compared the reconstruction performance of our proposed method with other optical diffraction tomography methods by means of numerical simulations. The results showed that our method provides RI and absorption images of 3D resolution targets with minimal artifacts. Finally, we apply the technique to imaging biological samples, e.g., the Thalassiosira diatoms, revealing the intrinsic structural diversity of the biological samples.
We present a 3D label-free refractive index (RI) imaging technique based on single-exposure intensity diffraction tomography (sIDT) utilizing a color-multiplexed illumination scheme. In our method, the chromatic LEDs corresponding R/G/B channels in an annular programmable array provide oblique illumination geometry which matches the numerical aperture of the objective precisely to maximize the spectrum coverage. A color intensity image encoding the scattering field of the specimen from different directions is captured, and monochromatic intensity images with respect to three color channels were separated and then used to recover the 3D RI distribution of the object following the process of IDT. In addition, the axial chromatic dispersion of focal lengths at different wavelengths introduced by the chromatic aberration of the objective lens and the spatial position misalignment of the ring LED source in the imaging system’s transfer functions modeling are both corrected to significantly reduce the artifacts in slice-based deconvolution procedure for the reconstruction of 3D RI distribution. Experimental results on MCF-7, Spirulina algae, and live C. elegans samples demonstrate the solid performance of the sIDT method in label-free, high-throughput, and real-time (∼ 24 fps) 3D volumetric biological imaging applications.
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