Single-shot quantitative phase imaging with polarization differential interference contrast
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Abstract
We present single-shot quantitative phase imaging with polarization differential interference contrast (PDIC) for a differential interference contrast microscope which records the unfiltered Stokes vector of the differential interference pattern with a polarization camera. PDIC enables high spatial resolution phase imaging in real-time, applicable to either absorptive or transparent samples and integrates simply with epi-fluorescence imaging. An algorithm with total variation regularization is further introduced to solve the quantitative phase map from the partial derivative along one single axis, improving the accuracy and the image quality from the Fourier transform solution. After quantifying the accuracy of PDIC phase imaging with numerical simulations and phantom measurements, we demonstrate the biomedical applications by imaging the quantitative phase of both stained and unstained histological tissue sections and visualizing the fission yeast Schizosaccharomyces pombe's cytokinesis.
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Live cell imaging of the fission yeast Schizosaccharomyces pombe at 3, 11, 22, 27, 30, and 34 mins in (a-f) (See Visualization I). This strain expresses the nuclear pore protein nup211 fused with the green fluorescent protein (nup211-GFP), marking the position of the nucleus. Arrowheads point to the septum, where cytokinesis occurs. Scale bar: 5µm.
It would be great to have a comparison to a standard DIC image here. It would help to answer questions like, is the image of the cleavage furrow substantially clearer in the PDIC image.
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The phase images for the stained and unstained serial cuts are overall similar. However, slight elevation of the phase is noticeable for the H&E stained section, especially in the areas surrounding some stromal regions.
The ability to image structural components of unstained material is very impressive. You have shown two examples of biological samples that are very thin. Have you looked at thicker samples? is there a limit to the thickness of samples that can be imaged?
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