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Visually precise, low-damage, single-cell spatial manipulation with single-pixel resolution.

ABSTRACT: The analysis of single living cells, including intracellular delivery and extraction, is essential for monitoring their dynamic biochemical processes and exploring intracellular heterogeneity. However, owing to the 2D view in bright-field microscopy and optical distortions caused by the cell shape and the variation in the refractive index both inside and around the cells, achieving spatially undistorted imaging for high-precision manipulation within a cell is challenging. Here, an accurate and visual system is developed for single-cell spatial manipulation by correcting the aberration for simultaneous bright-field triple-view imaging. Stereo information from the triple view enables higher spatial resolution that facilitates the precise manipulation of single cells. In the bright field, we resolved the spatial locations of subcellular structures of a single cell suspended in a medium and measured the random spatial rotation angle of the cell with a precision of ±5°. Furthermore, we demonstrated the visual manipulation of a probe to an arbitrary spatial point of a cell with an accuracy of <1 pixel. This novel system is more accurate and less destructive for subcellular content extraction and drug delivery.

SUBMITTER: Zhang Q

PROVIDER: S-EPMC8179525 | biostudies-literature | 2021 Feb

REPOSITORIES: biostudies-literature

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Visually precise, low-damage, single-cell spatial manipulation with single-pixel resolution.

Zhang Qi Q Shao Yunlong Y Li Boye B Wu Yuanyuan Y Dong Jingying J Zhang Dongtang D Wang Yanan Y Yan Yong Y Wang Xiayan X Pu Qiaosheng Q Guo Guangsheng G

Chemical science 20210202 11

The analysis of single living cells, including intracellular delivery and extraction, is essential for monitoring their dynamic biochemical processes and exploring intracellular heterogeneity. However, owing to the 2D view in bright-field microscopy and optical distortions caused by the cell shape and the variation in the refractive index both inside and around the cells, achieving spatially undistorted imaging for high-precision manipulation within a cell is challenging. Here, an accurate and v ...[more]

PMID: 34163682

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Project description:PurposePrevious efforts at increasing spatial resolution have relied on decreasing focal spot and or detector element size. Many "super resolution" methods require physical movement of a component of the imaging system. This work describes a method for achieving spatial resolution on a scale smaller than the detector pixel without motion of the object or detector.MethodsWe introduce a weighting of the photon energy spectrum on a length scale smaller than a single pixel using a physical filter that can be placed between the focal spot and the object, between the object and the detector, or integrated into the x-ray source or detector. We refer to the method as sub pixel encoding (SPE). We show that if one acquires multiple measurements (i.e. x-ray projections), information can be synthesized at a spatial scale defined by the spectrum modulation, not the detector element size. Specifically, if one divides a detector pixel into n sub regions, and m photon-matter interactions are present, the number of x-ray measurements needed to solve for the detector response of each sub region is mxn. We discuss realizations of SPE using multiple x-ray spectra with an energy integrating detector, a single spectra with a photon counting detector, and the single photon-matter interaction case. We demonstrate the feasibility of the approach using a simulated energy integrating detector with a detector pitch of 2 mm for 80-140 kV medical and 200-600 kV industrial applications. Phantoms used for both example SPE realization had some features only a 1 mm detector could resolve. We calculate the covariance matrix of SPE output to characterize the and noise propagation and correlation of our test examples.ResultsThe mathematical foundation of SPE is provided, with details worked out for several detector types and energy ranges. Two numerical simulations were provided to demonstrate feasibility. In both the medical and industrial simulations, some phantom features were only observable with the 1 mm and SPE synthesized 2 mm detector, while the 2 mm detector was not able to visualize them. Covariance matrix analysis demonstrated negative diagonal terms for both example cases.ConclusionsThe concept of encoding object information at a length scale smaller than a single pixel element, and then retrieving that information was introduced. SPE simultaneously allows for an increase in spatial resolution and provides "dual energy" like information about the underlying photon-matter interactions.

Dataset Information

Visually precise, low-damage, single-cell spatial manipulation with single-pixel resolution.

Publications

Visually precise, low-damage, single-cell spatial manipulation with single-pixel resolution.

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