Tomographic diffractive microscopy of living cells based on a common-path configuration

Hsu, Wei-Chen; Su, Jing-Wei; Tseng, Te-Yu; Sung, Kung-Bin

doi:10.1364/ol.39.002210

Cited by 64 publications

(35 citation statements)

References 16 publications

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“…All phase projections were then processed digitally to create the 3D refractive‐index map of the cell by both the filtered back projection and the diffraction‐theory reconstruction algorithms 13, 30. In this reconstruction process, each projection is mapped to a surface in the 3D Fourier space, where the full rotation provided by DEP enables a full angular coverage of the Fourier space, in contrast to previous methods possessing limited angular range1, 2, 3, 4, 5, 6, 7 (see comparison in Figure 1). …”

Section: Resultsmentioning

confidence: 99%

See 1 more Smart Citation

Rapid 3D Refractive‐Index Imaging of Live Cells in Suspension without Labeling Using Dielectrophoretic Cell Rotation

et al. 2016

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A major challenge in the field of optical imaging of live cells is achieving rapid, 3D, and noninvasive imaging of isolated cells without labeling. If successful, many clinical procedures involving analysis and sorting of cells drawn from body fluids, including blood, can be significantly improved. A new label‐free tomographic interferometry approach is presented. This approach provides rapid capturing of the 3D refractive‐index distribution of single cells in suspension. The cells flow in a microfluidic channel, are trapped, and then rapidly rotated by dielectrophoretic forces in a noninvasive and precise manner. Interferometric projections of the rotated cell are acquired and processed into the cellular 3D refractive‐index map. Uniquely, this approach provides full (360°) coverage of the rotation angular range around any axis, and knowledge on the viewing angle. The experimental demonstrations presented include 3D, label‐free imaging of cancer cells and three types of white blood cells. This approach is expected to be useful for label‐free cell sorting, as well as for detection and monitoring of pathological conditions resulting in cellular morphology changes or occurrence of specific cell types in blood or other body fluids.

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Section: Resultsmentioning

confidence: 99%

“…To view the sample from multiple angles, one can rotate the illumination beam, while leaving the measured specimen stationary 1, 2, 3, 4, 5, 6, 7. This approach is noninvasive to the sample during data acquisition.…”

Section: Introductionmentioning

confidence: 99%

Rapid 3D Refractive‐Index Imaging of Live Cells in Suspension without Labeling Using Dielectrophoretic Cell Rotation

et al. 2016

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“…Refractive-index detection offers techniques to noninvasively probe the structural and chemical information of biological cells including their morphology, dynamics and concentration of specific substances [2][3][4][5][6][7][8][9].…”

Section: Introductionmentioning

confidence: 99%

“…Previous approaches measured the transmitted field from multiple angles, as implemented by either illumination beam rotation [4][5][6][7][8] or sample rotation [2][3]9]. The main concept of the first method is to rotate the illumination beam, while the specimen and optical setup are fixed.…”

Section: Introductionmentioning

confidence: 99%

Tomographic phase microscopy using optical tweezers

et al. 2015

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We review our technique for tomographic phase microscopy with optical tweezers [1]. This tomographic phase microscopy approach enables full 3-D refractive-index reconstruction. Tomographic phase microscopy measures quantitatively the 3-D distribution of refractive-index in biological cells. We integrated our external interferometric module with holographic optical tweezers for obtaining quantitative phase maps of biological samples from a wide range of angles. The close-tocommon-path, off-axis interferometric system enables a full-rotation tomographic acquisition of a single cell using holographic optical tweezers for trapping and manipulating with a desired array of traps, while acquiring phase information of a single cell from all different angles and maintaining the native surrounding medium. We experimentally demonstrated two reconstruction algorithms: the filtered back-projection method and the Fourier diffraction method for 3-D refractive index imaging of yeast cells.

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“…Previously, ODT has been widely used to study various biological samples including red blood cells [15][16][17][18][19][20][21][22] , white blood cells (WBC) 23,24 , hepatocytes 25 , cancer cells 16,[26][27][28][29][30][31][32] , neurons 32,33 , bacteria 34,35 , phytoplankton 36 , and hair 37 . In our previous study, we reported that ODT enables the quantitative analysis of WBCs including lymphocytes and macrophages 23 .…”

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confidence: 99%

Label-free identification of non-activated lymphocytes using three-dimensional refractive index tomography and machine learning

Yoon

Kim

et al. 2017

Preprint

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Identification of lymphocyte cell types is crucial for understanding their pathophysiologic roles in human diseases.Current methods for discriminating lymphocyte cell types primarily relies on labelling techniques with magnetic beads or fluorescence agents, which take time and have costs for sample preparation and may also have a potential risk of altering cellular functions. Here, we present label-free identification of non-activated lymphocyte subtypes using refractive index tomography. From the measurements of three-dimensional refractive index maps of individual lymphocytes, the morphological and biochemical properties of the lymphocytes are quantitatively retrieved. Machine learning methods establish an optimized classification model using the retrieved quantitative characteristics of the lymphocytes to identify lymphocyte subtypes at the individual cell level. We show that our approach enables label-free identification of three lymphocyte cell types (B, CD4+ T, and CD8+ T lymphocytes) with high specificity and sensitivity. The present method will be a versatile tool for investigating the pathophysiological roles of lymphocytes in various diseases including cancers, autoimmune diseases, and virus infections.Lymphocytes consisting of various cell types including B, helper (CD4+) T, cytotoxic (CD8+) T, and regulatory T lymphocytes, and play crucial roles in the adaptive immune system 1 . Each lymphocyte cell type has different functions: B lymphocytes produce antibodies, and T lymphocytes recognize a specific antigen and execute effector functions. The lymphocyte population and function are tightly regulated to defend the host against harmful invaders or abnormal conditions 1,2 . Disturbances in lymphocyte function and regulation are related to various diseases including cancers 3-5 , autoimmune diseases 6,7 , and virus infections 8,9 .. CC-BY 4.0 International license not peer-reviewed) is the author/funder. It is made available under a The copyright holder for this preprint (which was . http://dx.doi.org/10.1101/107805 doi: bioRxiv preprint first posted online Feb. 11, 2017; To understand the roles of different types of lymphocytes, several methods based on labelling techniques have been developed to identify and discriminate lymphocyte cell types. Because different kinds of lymphocytes have very similar cellular morphology such as a large nucleus with small cytosolic regions and round shapes, conventional optical methods such as bright-field microscopy or phase contrast microscopy have limited ability in classifying lymphocyte cell types 10 .To overcome this, a specific surface membrane proteins, known as surface markers, are recognized and tagged with magnetic beads or fluorescence molecules via antigen-antibody binding. And then, specific types of lymphocytes are identified and separated by magnetic forces or fluorescence signals 11 . Targeting surface markers is a precise and efficient manner to determine the cell types; however, labelling methods have potential risks of altering cellular functions b...

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Tomographic diffractive microscopy of living cells based on a common-path configuration

Cited by 64 publications

References 16 publications

Rapid 3D Refractive‐Index Imaging of Live Cells in Suspension without Labeling Using Dielectrophoretic Cell Rotation

Rapid 3D Refractive‐Index Imaging of Live Cells in Suspension without Labeling Using Dielectrophoretic Cell Rotation

Tomographic phase microscopy using optical tweezers

Label-free identification of non-activated lymphocytes using three-dimensional refractive index tomography and machine learning

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