scholarly journals Dynamic Effective Elasticity of Melanoma Cells Under Shear and Elongational Flow Confirms Estimation From Force Spectroscopy

Author(s):  
Anna Martina Jötten ◽  
Simon Neidinger ◽  
Julia K. Tietze ◽  
Julia Welzel ◽  
Christoph Westerhausen

The detection and enrichment of circulating melanoma cells is a challenge as the cells are very heterogeneous in terms of their biomechanical properties and surface markers. In addition, there is a lack of valid and reliable biomarkers that predict progress and therapeutic response. We here analyzed the elasticity of A375 melanoma cells applying force spectroscopy and a microfluidic method. To identify and eventually separate circulating tumor cells, it is crucial to precisely know their physical properties. First, we used standard AFM force spectroscopy, where the elasticity of the cells is calculated from indentation with a pyramidal tip. To extend the limits of measurement with a tip, we then used cantilevers without a tip to press on the cells over a large area. The resulting Young’s moduli are slightly lower and vary less without tip presumably because of the inhomogeneity of the cells. Finally, we implemented our microfluidic method. We measured single cell elasticity by analyzing its deformation in high-speed micrographs while passing a stenosis. Combining the force field and the change in shape provides the basis for a stress strain diagram. The results from microfluidic deformation analysis were in accordance with the results from force spectroscopy. The microfluidic method provides advantages over conventional methods, since it is less invasive and less likely to harm the cell during the measurement, and the whole cell is measured as one entity without contact to a stiff substrate, while force spectroscopy is limited to the contact area of the tip, and in some cases dependent of the cell substrate interaction. Consequently, microfluidic deformation analysis allows to predict the overall elastic behavior of the whole inhomogeneous cell in three-dimensional force fields. This method may contribute to improve the detection of circulation melanoma cells in the clinical practice.

Biophysica ◽  
2021 ◽  
Vol 1 (4) ◽  
pp. 445-457
Author(s):  
Anna Martina Jötten ◽  
Simon V. Neidinger ◽  
Julia K. Tietze ◽  
Julia Welzel ◽  
Christoph Westerhausen

The detection and enrichment of circulating melanoma cells is a challenge, as the cells are very heterogeneous in terms of their biomechanical properties and surface markers. In addition, there is a lack of valid and reliable biomarkers predicting progress and therapeutic response. In this study, we analyze the elasticity of A375 melanoma cells by applying force spectroscopy and a microfluidic method. To identify and eventually separate freely circulating tumor cells, it is crucial to know their physical properties precisely. First, we use standard AFM force spectroscopy, where the elasticity of the cells is calculated from indentation with a pyramidal tip. To extend the limits of the measurements with a tip, we then use cantilevers without a tip to apply force over a larger area of the cells. The resulting Young’s moduli are slightly lower and vary less without the tip, presumably because of the spatial inhomogeneity of the cells. Finally, we implement our microfluidic method: we measure single cell elasticity by analyzing their deformation in high-speed micrographs while passing a stenosis. Combining the force field and the change in shape provides the basis for a stress–strain diagram. The results from the microfluidic deformation analysis were well in accordance with the results from force spectroscopy. The microfluidic method, however, provides advantages over conventional methods, as it is less invasive and less likely to harm the cell during the measurement. The whole cell is measured as one entity without having contact to a stiff substrate, while force spectroscopy is limited to the contact area of the tip, and in some cases dependent of the cell substrate interaction. Consequently, microfluidic deformation analysis allows us to predict the overall elastic behavior of the whole, inhomogeneous cell in three-dimensional force fields. This method may contribute to improve the detection of circulating melanoma cells in the clinical practice.


2016 ◽  
Vol 138 (3) ◽  
Author(s):  
Simon Heide-Jørgensen ◽  
Sellaswasmy Kumaran Krishna ◽  
Jonas Taborsky ◽  
Tommy Bechsgaard ◽  
Rachid Zegdi ◽  
...  

The transcatheter aortic valve implantation (TAVI) valve is a bioprosthetic valve within a metal stent frame. Like traditional surgical bioprosthetic valves, the TAVI valve leaflet tissue is expected to calcify and degrade over time. However, clinical studies of TAVI valve longevity are still limited. In order to indirectly assess the longevity of TAVI valves, an estimate of the mechanical wear and tear in terms of valvular deformation and strain of the leaflets under various conditions is warranted. The aim of this study was, therefore, to develop a platform for noncontact TAVI valve deformation analysis with both high temporal and spatial resolutions based on stereophotogrammetry and digital image correlation (DIC). A left-heart pulsatile in vitro flow loop system for mounting of TAVI valves was designed. The system enabled high-resolution imaging of all three TAVI valve leaflets simultaneously for up to 2000 frames per second through two high-speed cameras allowing three-dimensional analyses. A coating technique for applying a stochastic pattern on the leaflets of the TAVI valve was developed. The technique allowed a pattern recognition software to apply frame-by-frame cross correlation based deformation measurements from which the leaflet motions and the strain fields were derived. The spatiotemporal development of a very detailed strain field was obtained with a 0.5 ms time resolution and a spatial resolution of 72 μm/pixel. Hence, a platform offering a new and enhanced supplementary experimental evaluation of tissue valves during various conditions in vitro is presented.


2011 ◽  
Vol 189-193 ◽  
pp. 1986-1990
Author(s):  
De Ping Liu ◽  
Jie Li ◽  
Yu Feng Su ◽  
Yu Ping Wang

Taking the high-speed CX series vertical milling compound machining center of CX8075 produced by Anyang Xinsheng Machine Tool Co., Ltd. as example, the machine three-dimensional simplified model is established, the source of the heat and the distribution of the important hot-points are analyzed, the machine temperature field distribution is derived which lays a foundation for the thermal error compensation. Taking into account the moving part-saddle of the machining center, its mathematic model is obtained, the important hot-points are studied, the thermodynamic parameters are determined. Based on ANSYS finite element method, the steady-state temperature field and the thermal deformation of saddle are presented, the optimal design of high-speed and high-accuracy machine tool is doned and its thermal deformation analysis is realized.


Author(s):  
Robert W. Mackin

This paper presents two advances towards the automated three-dimensional (3-D) analysis of thick and heavily-overlapped regions in cytological preparations such as cervical/vaginal smears. First, a high speed 3-D brightfield microscope has been developed, allowing the acquisition of image data at speeds approaching 30 optical slices per second. Second, algorithms have been developed to detect and segment nuclei in spite of the extremely high image variability and low contrast typical of such regions. The analysis of such regions is inherently a 3-D problem that cannot be solved reliably with conventional 2-D imaging and image analysis methods.High-Speed 3-D imaging of the specimen is accomplished by moving the specimen axially relative to the objective lens of a standard microscope (Zeiss) at a speed of 30 steps per second, where the stepsize is adjustable from 0.2 - 5μm. The specimen is mounted on a computer-controlled, piezoelectric microstage (Burleigh PZS-100, 68/μm displacement). At each step, an optical slice is acquired using a CCD camera (SONY XC-11/71 IP, Dalsa CA-D1-0256, and CA-D2-0512 have been used) connected to a 4-node array processor system based on the Intel i860 chip.


2021 ◽  
Author(s):  
Scott J. Peltier ◽  
Brian E. Rice ◽  
Ethan Johnson ◽  
Venkateswaran Narayanaswamy ◽  
Marvin E. Sellers

2018 ◽  
Author(s):  
Yi Chen Mazumdar ◽  
Michael E. Smyser ◽  
Jeffery Dean Heyborne ◽  
Daniel Robert Guildenbecher

2020 ◽  
Vol 11 (1) ◽  
Author(s):  
Jiang Lan Fan ◽  
Jose A. Rivera ◽  
Wei Sun ◽  
John Peterson ◽  
Henry Haeberle ◽  
...  

AbstractUnderstanding the structure and function of vasculature in the brain requires us to monitor distributed hemodynamics at high spatial and temporal resolution in three-dimensional (3D) volumes in vivo. Currently, a volumetric vasculature imaging method with sub-capillary spatial resolution and blood flow-resolving speed is lacking. Here, using two-photon laser scanning microscopy (TPLSM) with an axially extended Bessel focus, we capture volumetric hemodynamics in the awake mouse brain at a spatiotemporal resolution sufficient for measuring capillary size and blood flow. With Bessel TPLSM, the fluorescence signal of a vessel becomes proportional to its size, which enables convenient intensity-based analysis of vessel dilation and constriction dynamics in large volumes. We observe entrainment of vasodilation and vasoconstriction with pupil diameter and measure 3D blood flow at 99 volumes/second. Demonstrating high-throughput monitoring of hemodynamics in the awake brain, we expect Bessel TPLSM to make broad impacts on neurovasculature research.


Nanomaterials ◽  
2021 ◽  
Vol 11 (4) ◽  
pp. 956
Author(s):  
Philipp Taus ◽  
Adrian Prinz ◽  
Heinz D. Wanzenboeck ◽  
Patrick Schuller ◽  
Anton Tsenov ◽  
...  

Biomimetic structures such as structural colors demand a fabrication technology of complex three-dimensional nanostructures on large areas. Nanoimprint lithography (NIL) is capable of large area replication of three-dimensional structures, but the master stamp fabrication is often a bottleneck. We have demonstrated different approaches allowing for the generation of sophisticated undercut T-shaped masters for NIL replication. With a layer-stack of phase transition material (PTM) on poly-Si, we have demonstrated the successful fabrication of a single layer undercut T-shaped structure. With a multilayer-stack of silicon oxide on silicon, we have shown the successful fabrication of a multilayer undercut T-shaped structures. For patterning optical lithography, electron beam lithography and nanoimprint lithography have been compared and have yielded structures from 10 µm down to 300 nm. The multilayer undercut T-shaped structures closely resemble the geometry of the surface of a Morpho butterfly, and may be used in future to replicate structural colors on artificial surfaces.


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