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Gradient light interference microscopy for 3D imaging of unlabeled specimens

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Multiple scattering limits the contrast in optical imaging of thick specimens. Here, we present gradient light interference microscopy (GLIM) to extract three-dimensional information from both thin and thick unlabeled specimens.… Click to show full abstract

Multiple scattering limits the contrast in optical imaging of thick specimens. Here, we present gradient light interference microscopy (GLIM) to extract three-dimensional information from both thin and thick unlabeled specimens. GLIM exploits a special case of low-coherence interferometry to extract phase information from the specimen, which in turn can be used to measure cell mass, volume, surface area, and their evolutions in time. Because it combines multiple intensity images that correspond to controlled phase shifts between two interfering waves, gradient light interference microscopy is capable of suppressing the incoherent background due to multiple scattering. GLIM can potentially become a valuable tool for in vitro fertilization, where contrast agents and fluorophores may impact the viability of the embryo. Since GLIM is implemented as an add-on module to an existing inverted microscope, we anticipate that it will be adopted rapidly by the biological community.Challenges in biological imaging include labeling, photobleaching and phototoxicity, as well as light scattering. Here, Nguyen et al. develop a quantitative phase method that uses low-coherence interferometry for label-free 3D imaging in scattering tissue.

Keywords: light interference; microscopy; unlabeled specimens; interference microscopy; gradient light

Journal Title: Nature Communications
Year Published: 2017

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