Video-rate tomographic phase microscopy

Fang-Yen, Christopher; Choi, Wonshik; Sung, Yongjin; Holbrow, Charles J.; Dasari, Ramachandra R.; Feld, Michael S.

doi:10.1117/1.3522506

Cited by 49 publications

(39 citation statements)

References 18 publications

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“…The experimental setup is equipped with a two-axis galvanometer mirror (GM, GVS012/M, Thorlabs, USA), which varies the illumination angle of an incident beam for the tomographic measurement. This setup is conceptually comparable with setups used in previous works [24,28].…”

Section: Optical Setupsupporting

confidence: 66%

High-Resolution 3-D Refractive Index Tomography and 2-D Synthetic Aperture Imaging of Live Phytoplankton

Lee¹,

K²,

Mubarok³

et al. 2014

Journal of the Optical Society of Korea

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Optical measurements of the morphological and biochemical imaging of phytoplankton are presented. Employing quantitative phase imaging techniques, 3-D refractive index maps and high-resolution 2-D quantitative phase images of individual live phytoplankton are simultaneously obtained without exogenous labeling agents. In addition, biochemical information of individual phytoplankton including volume, mass, and density of individual phytoplankton are also quantitatively obtained from the measured refractive index distributions. We expect the present method to become a powerful tool for the study of phytoplankton.

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Section: Optical Setupsupporting

confidence: 66%

High-Resolution 3-D Refractive Index Tomography and 2-D Synthetic Aperture Imaging of Live Phytoplankton

Lee¹,

K²,

Mubarok³

et al. 2014

Journal of the Optical Society of Korea

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“…In general, determination of the 3D structure of an object from the knowledge of the scattered optical fields requires a weakly scattering object (55,56). This assumption also forms the basis of our technique in recovering the 3D scattering potential of the objects similar to existing optical tomography platforms (34)(35)(36)(37)(38)(39)(40)(41). Therefore, as further detailed in SI Text, the bulk of the photons incident on an object should encounter at most a single scattering event before being detected in our transmission holographic imaging geometry.…”

Section: Discussionmentioning

confidence: 97%

“…S7, S9, and S10) is a direct outcome of our lens-free holographic recording geometry, which is not restricted by the limited DOF of high NA objective lenses as utilized in some other tomography or holography schemes (36)(37)(38)(39)(40)(41)(42)(43). Lens-free holographic recording at multiple angles enables digital reconstruction of projection images at any given depth of interest, around which tomograms can be computed.…”

Section: Discussionmentioning

confidence: 99%

“…In recent years, there has also been an increased interest in optical imaging modalities that enable sectional imaging (34)(35)(36)(37)(38)(39)(40)(41)(42)(43)(44)(45)(46)(47)(48). As an example, optical projection tomography (OPT) (34), where an optically cleared specimen immersed in index-matching gel is rotated with respect to the fixed optical path of a conventional lens-based microscope, offers an isotropic resolution of approximately 10 μm in all three dimensions within an imaging volume of up to approximately 1 cm 3 .…”

mentioning

confidence: 99%

“…A modified version of OPT by using high NA objective lenses has also been implemented (35) recently to achieve submicron resolution cell imaging over a significantly reduced volume of, e.g., <0.0005 mm 3 . Optical diffraction tomography (ODT) is another powerful technique where digital holography is utilized to reconstruct the 3D refractive index distribution of the specimen by changing the illumination direction (36)(37)(38)(39)(40), rotating the object (41), or by capturing multiple images at different wavelengths (42,43). These tomographic systems can routinely image cells potentially achieving submicron resolution in all three dimensions.…”

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confidence: 99%

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Lens-free optical tomographic microscope with a large imaging volume on a chip

Isikman

Bishara²,

Mavandadi³

et al. 2011

Proc. Natl. Acad. Sci. U.S.A.

203

213

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We present a lens-free optical tomographic microscope, which enables imaging a large volume of approximately 15 mm 3 on a chip, with a spatial resolution of <1 μm× < 1 μm× < 3 μm in x, y and z dimensions, respectively. In this lens-free tomography modality, the sample is placed directly on a digital sensor array with, e.g., ≤4 mm distance to its active area. A partially coherent light source placed approximately 70 mm away from the sensor is employed to record lens-free in-line holograms of the sample from different viewing angles. At each illumination angle, multiple subpixel shifted holograms are also recorded, which are digitally processed using a pixel superresolution technique to create a single high-resolution hologram of each angular projection of the object. These superresolved holograms are digitally reconstructed for an angular range of AE50°, which are then back-projected to compute tomograms of the sample. In order to minimize the artifacts due to limited angular range of tilted illumination, a dual-axis tomography scheme is adopted, where the light source is rotated along two orthogonal axes. Tomographic imaging performance is quantified using microbeads of different dimensions, as well as by imaging wild-type Caenorhabditis elegans. Probing a large volume with a decent 3D spatial resolution, this lens-free optical tomography platform on a chip could provide a powerful tool for high-throughput imaging applications in, e.g., cell and developmental biology. L ight microscopy has been an irreplaceable tool in life sciences for several centuries. The quest to resolve smaller features with better resolution and contrast has improved the capabilities of this important tool at the cost of relatively increasing its size and complexity (1). On the other hand, we have experienced the flourishing of emerging technologies such as microfluidic and lab-on-a-chip systems, which offer fast and efficient handling and processing of biological samples within highly miniaturized architectures (2-7). The optical inspection of the specimen, however, is still being performed by conventional light microscopes, which has in general several orders of magnitude size mismatch compared to the scale of the microfluidic systems. As a result, there is a clear need for alternative compact microscopy modalities toward integration with miniaturized lab-on-a-chip platforms (8).The push for new optical microscopy modalities is not solely driven by the need for miniaturization and microfluidic integration. The fact that high resolution is achieved at the cost of significant field-of-view (FOV) reduction is another fundamental limitation of lens-based imaging. The relatively small FOV of conventional light microscopy brings additional challenges for its application to several important problems such as rare cell imaging or optical phenotyping of model organisms (9-15), where high-throughput microscopy is highly desired.In order to provide complementary solutions to these aforementioned needs, several lens-free digital microscopy techniqu...

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Digital holographic microtomography for high‐resolution refractive index mapping of live cells

Hsu

Chou

et al. 2012

Journal of Biophotonics

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Quantification of three-dimensional (3D) refractive index (RI) with sub-cellular resolution is achieved by digital holographic microtomography (DHμT) using quantitative phase images measured at multiple illumination angles. The DHμT system achieves sensitive and fast phase measurements based on iterative phase extraction algorithm and asynchronous phase shifting interferometry without any phase monitoring or active control mechanism. A reconstruction algorithm, optical diffraction tomography with projection on convex sets and total variation minimization, is implemented to substantially reduce the number of angular scattered fields needed for reconstruction without sacrificing the accuracy and quality of the reconstructed 3D RI distribution. Tomogram of a living CA9-22 cell is presented to demonstrate the performance of the method. Further, a statistical analysis of the average RI of the nucleoli, the nucleus excluding the nucleoli and the cytoplasm of twenty CA9-22 cells is performed.

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Video-rate tomographic phase microscopy

Cited by 49 publications

References 18 publications

High-Resolution 3-D Refractive Index Tomography and 2-D Synthetic Aperture Imaging of Live Phytoplankton

High-Resolution 3-D Refractive Index Tomography and 2-D Synthetic Aperture Imaging of Live Phytoplankton

Lens-free optical tomographic microscope with a large imaging volume on a chip

Digital holographic microtomography for high‐resolution refractive index mapping of live cells

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