Limited-angle tomographic phase microscopy utilizing confocal scanning fluorescence microscopy

We present a multimodal imaging technique, combining tomographic phase microscopy with limited angular projection range and number, and two-channel spinning-disk confocal scanning fluorescence microscopy. This technique allows high-accuracy 3D refractive index (RI) profiling of live cells in spite of the missing projections. The cellular outer shape and its interior organelles measured by the confocal fluorescence imaging not only specify the cell in molecular levels, but also provide the 3D distributions of the whole cell as well as its organelles. We take these additional 3D morphological details as constraints in Gerchberg-Papoulis-based optical diffraction tomography algorithm. We then obtain an accurate 3D RI tomogram, even with a sparse angular range having a small number of perspective projections, otherwise providing low-accuracy RI reconstruction. Then, we obtain both cellular molecular specificity and inner RI values of the cell and its organelles. We compare the reconstructed 3D RI profiles of various samples, demonstrating the superiority of the proposed technique.

Automated summary

The proposed multimodal imaging technique combines tomographic phase microscopy with limited angular projection range and two-channel spinning-disk confocal scanning fluorescence microscopy. It allows high-accuracy 3D refractive index profiling of live cells, providing both cellular morphology and molecular specificity. The additional 3D morphological details act as constraints to improve the accuracy of RI reconstruction, demonstrating the superiority of the technique in various samples.