Focus stabilization by axial position feedback in biomedical imaging microscopy

Lee, Seohyun; Kim, Hyuno; Higuchi, Hideo

doi:10.1109/sas.2018.8336767

Cited by 9 publications

(8 citation statements)

References 13 publications

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“…In the presented work, we showed precise vesicle movement trajectory in a complex microtubule network structure in a living cell by analyzing the three-dimensional movement trajectory data of vesicles acquired at a high spatiotemporal precision, achieved by highly stable microscope stage with position feedback system (22). Particularly, we discovered rotational movement of vesicles around the axis of the microtubule, which consists of characteristic quick dodging and slow walking.…”

Section: Discussionmentioning

confidence: 90%

“…The specimen was loaded on the microscope stage and kept at the physiological temperature of 37°C by a heater (Tokai HIT) mounted on the stage. For the stable imaging, a capacitive sensor-based stage stabilization system (22) was attached to achieve high spatial resolution (near 4 nm) in terms of the standard deviation for each x, y, and z coordinate. The dual-focus optics was built by a customized optical path setting, to collect the identical point source at two different focal planes, which were separated by 1 µm in the axial direction.…”

Section: Live-cell Imagingmentioning

confidence: 99%

“…In the present work, we report the detailed features of vesicle movement on a complicated 3D microtubules network structure at high spatiotemporal precision ( Fig. S1), achieved by our microscope stage position feedback system (22) that guarantees approximately 3 nm of x, y and 5 nm of axial spatial precision at 100 frames per second. First, the 3D angles between adjacent microtubules where the vesicle sequentially interacted are calculated based on our novel numerical analysis method reported previously (21).…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

3D rotational motion of an endocytic vesicle on a complex microtubule network in a living cell

Lee

Higuchi

2019

Preprint

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The transport dynamics of endocytic vesicles in a living cell contains essential biomedical information. Although the movement mechanism of a vesicle by motor proteins has been revealed, understanding the precise movement of vesicles on the cytoskeleton in a living cell has been considered challenging, due to the complex 3D network of cytoskeletons. Here, we specify the shape of the 3D interaction between the vesicle and microtubule, based on the theoretically estimated location of the microtubule and the vesicle trajectory data acquired at high spatial and temporal precision. We detected that vesicles showed more frequent direction changes with either in very acute or in obtuse angles than right angles, on similar time scales in a microtubule network. Interestingly, when a vesicle interacted with a relatively longer (> 400 nm) microtubule filament, rotational movement along the axis of the microtubule was frequently observed, implying an obstacle-avoiding motion. Our results are expected to give in-depth insight into understanding the actual 3D interactions between the intracellular molecule and complex cytoskeletal network.

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Section: Discussionmentioning

confidence: 90%

Section: Live-cell Imagingmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

3D rotational motion of an endocytic vesicle on a complex microtubule network in a living cell

Lee

Higuchi

2019

Preprint

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show abstract

“…The specimen was loaded on the microscope stage and kept at the physiological temperature of 37°C by a heater (IN-ONI-F1, Tokai HIT, Japan) mounted on the stage. For the stable imaging, a capacitive sensor-based stage stabilization system [20] was attached to achieve high spatial resolution (near 4 nm) in terms of the standard deviation for each x, y, and z coordinate. The dual-focus optics was built by a customized optical path setting, to collect the identical point source at two different focal planes, which were separated by 1 µm in the axial direction.…”

Section: Live-cell Imagingmentioning

confidence: 99%

“…In the present work, we report the detailed features of vesicle movement on a complicated 3D microtubules network structure at high spatiotemporal precision , achieved by our microscope stage position feedback system [20] that guarantees approximately 3 nm of x, y and 5 nm of axial spatial precision at 100 frames per second. First, the 3D angles between adjacent microtubules where the vesicle sequentially interacted are calculated based on our novel numerical analysis method reported previously [21].…”

Section: Introductionmentioning

confidence: 99%

3D rotational motion of an endocytic vesicle on a complex microtubule network in a living cell

Lee

Higuchi

2019

Biomed. Opt. Express

Self Cite

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The transport dynamics of endocytic vesicles in a living cell contains essential biomedical information. Although the movement mechanism of a vesicle by motor proteins has been revealed, understanding the precise movement of vesicles on the cytoskeleton in a living cell has been considered challenging, due to the complex 3D network of cytoskeletons. Here, we specify the shape of the 3D interaction between the vesicle and microtubule, based on the theoretically estimated location of the microtubule and the vesicle trajectory data acquired at high spatial and temporal precision. We detected that vesicles showed more frequent direction changes with either in very acute or in obtuse angles than right angles, on similar time scales in a microtubule network. Interestingly, when a vesicle interacted with a relatively longer (> 400 nm) microtubule filament, rotational movement along the axis of the microtubule was frequently observed. Our results are expected to give in-depth insight into understanding the actual 3D interactions between the intracellular molecule and complex cytoskeletal network.

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A 3-DOF multi-depth-of-field carrier automatic focusing stage

Zhong,

Nie,

Liao

et al. 2024

Smart Mater. Struct.

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Automatic focusing Stage is an important part of automatic focusing system, a 3-DOF (z-axis displacement and XY axis tilt) Multi-Depth-Of-Field Carrier Automatic Focusing Stage (CAFS) is designed, manufactured and tested in this paper. The stage is driven by four piezoelectric (PZT) actuators. Based on four bridge amplifying mechanisms, large displacement travel and tilt Angle are obtained. Compared with other focusing stages, the developed stage adopts the focusing mode of directly adjusting the spatial position of the sample to realize focusing, which not only avoids the inherent defects of objective focusing mode, but also has the advantages of compact structure, low shape and overall manufacturing. In this paper, the structural parameters and the overall design of the stage are determined, and then the characteristics of the focusing stage are finite element analyzed (FEA). Finally, the performance of the focusing stage is studied experimentally. The maximum Z-axis output displacement of the focusing stage is 213.5μm, and the maximum x-direction and Y-direction tilt angles are 1.5 and 1.7mrad, respectively. For the application of CAFS, the practicability of CAFS is verified by Z-axis image superposition acquisition method in automatic focusing system.

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Focus stabilization by axial position feedback in biomedical imaging microscopy

Cited by 9 publications

References 13 publications

3D rotational motion of an endocytic vesicle on a complex microtubule network in a living cell

3D rotational motion of an endocytic vesicle on a complex microtubule network in a living cell

3D rotational motion of an endocytic vesicle on a complex microtubule network in a living cell

A 3-DOF multi-depth-of-field carrier automatic focusing stage

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