scholarly journals Quantifying Molecular Forces with Serially Connected Force Sensors

2018 ◽  
Author(s):  
Y. Murad ◽  
I. T.S. Li
Keyword(s):  
Cell Adhesion ◽  
Dynamic Range ◽  
Computational Study ◽  
Experimental Studies ◽  
High Signal ◽  
Force Sensors ◽  
Adhesion Forces ◽  
Receptor Ligand ◽  
Sequence Composition ◽  
Molecular Force

AbstractTo understand the mechanical forces involved in cell adhesion, molecular force sensors have been developed to study tension through adhesion proteins. Recently, a class of molecular force sensors called tension gauge tether (TGT) have been developed that rely on irreversible force-dependent dissociation of DNA duplex to study cell adhesion forces. While the TGT offer high signal-to-noise ratio and is ideal for studying fast / single molecular adhesion processes, quantitative interpretation of experimental results has been challenging. Here we used computational approach to investigate how TGT fluorescence readout can be quantitatively interpreted. In particular we studied force sensors made of a single TGT, multiplexed single TGTs, and two TGTs connected in series. Our results showed that fluorescence readout using a single TGT can result from drastically different combinations of force history and adhesion event density that span orders of magnitude. In addition, the apparent behaviour of the TGT is influenced by the tethered receptor-ligand, making it necessary to calibrate the TGT with every new receptor-ligand. To solve this problem, we proposed a system of two serially connected TGTs. Our result shows that not only is the ratiometric readout of serial TGT independent of the choice of receptor-ligand, it is able to reconstruct force history with sub-pN force resolution. This is also not possible by simply multiplexing different types of TGTs together. Lastly, we systematically investigated how sequence composition of the two serially connected TGTs can be tuned to achieve different dynamic range. This computational study demonstrated how serially connected irreversible molecular dissociation processes can accurately quantify molecular force, and laid the foundation for subsequent experimental studies.

scholarly journals Quantification of fast molecular adhesion by fluorescence footprinting

2021 ◽  
Vol 7 (34) ◽  
pp. eabe6984
Author(s):  
Adam B. Yasunaga ◽  
Isaac T. S. Li
Keyword(s):  
Shear Stress ◽  
Dynamic Range ◽  
Force Distribution ◽  
Receptor Ligand ◽  
Ligand Dissociation ◽  
Molecular Adhesion ◽  
Molecular Force ◽  
Nonequilibrium Conditions ◽  
Adhesion Complex

Rolling adhesion is a unique process in which the adhesion events are short-lived and operate under highly nonequilibrium conditions. These characteristics pose a challenge in molecular force quantification, where in situ measurement of these forces cannot be achieved with molecular force sensors that probe near equilibrium. Here, we demonstrated a quantitative adhesion footprint assay combining DNA-based nonequilibrium force probes and modeling to measure the molecular force involved in fast rolling adhesion. We were able to directly profile the ensemble molecular force distribution in our system during rolling adhesion with a dynamic range between 0 and 18 pN. Our results showed that the shear stress driving bead rolling motility directly controls the molecular tension on the probe-conjugated adhesion complex. Furthermore, the shear stress can steer the dissociation bias of components within the molecular force probe complex, favoring either DNA probe dissociation or receptor-ligand dissociation.

scholarly journals Ab initiostructure determination of nanocrystals of organic pharmaceutical compounds by electron diffraction at room temperature using a Timepix quantum area direct electron detector

2016 ◽  
Vol 72 (2) ◽  
pp. 236-242 ◽  
Author(s):  
E. van Genderen ◽  
M. T. B. Clabbers ◽  
P. P. Das ◽  
A. Stewart ◽  
I. Nederlof ◽  
...  
Keyword(s):  
Electron Diffraction ◽  
Room Temperature ◽  
Dynamic Range ◽  
Signal To Noise Ratio ◽  
Three Dimensional ◽  
Direct Methods ◽  
Liquid Nitrogen Temperature ◽  
High Dynamic Range ◽  
Electron Diffraction Data ◽  
High Signal

Until recently, structure determination by transmission electron microscopy of beam-sensitive three-dimensional nanocrystals required electron diffraction tomography data collection at liquid-nitrogen temperature, in order to reduce radiation damage. Here it is shown that the novel Timepix detector combines a high dynamic range with a very high signal-to-noise ratio and single-electron sensitivity, enablingab initiophasing of beam-sensitive organic compounds. Low-dose electron diffraction data (∼0.013 e− Å−2 s−1) were collected at room temperature with the rotation method. It was ascertained that the data were of sufficient quality for structure solution using direct methods using software developed for X-ray crystallography (XDS,SHELX) and for electron crystallography (ADT3D/PETS,SIR2014).

scholarly journals Model Calibration for a Rigid Hexapod-Based End-Effector with Integrated Force Sensors

Sensors ◽  
2021 ◽  
Vol 21 (10) ◽  
pp. 3537
Author(s):  
Christian Friedrich ◽  
Steffen Ihlenfeldt
Keyword(s):  
Parameter Identification ◽  
Degrees Of Freedom ◽  
Force Measurement ◽  
Experimental Studies ◽  
Measurement Model ◽  
Calibration Procedure ◽  
Force Sensors ◽  
End Effector ◽  
Measurement Models ◽  
Fast Procedure

Integrated single-axis force sensors allow an accurate and cost-efficient force measurement in 6 degrees of freedom for hexapod structures and kinematics. Depending on the sensor placement, the measurement is affected by internal forces that need to be compensated for by a measurement model. Since the parameters of the model can change during machine usage, a fast and easy calibration procedure is requested. This work studies parameter identification procedures for force measurement models on the example of a rigid hexapod-based end-effector. First, measurement and identification models are presented and parameter sensitivities are analysed. Next, two excitation strategies are applied and discussed: identification from quasi-static poses and identification from accelerated continuous trajectories. Both poses and trajectories are optimized by different criteria and evaluated in comparison. Finally, the procedures are validated by experimental studies with reference payloads. In conclusion, both strategies allow accurate parameter identification within a fast procedure in an operational machine state.

Essential arterial hypertension patients present higher cell adhesion forces, contributing to fibrinogen-dependent cardiovascular risk

Nanoscale ◽  
2017 ◽  
Vol 9 (39) ◽  
pp. 14897-14906 ◽  
Author(s):  
Ana F. Guedes ◽  
Filomena A. Carvalho ◽  
Carlos Moreira ◽  
José B. Nogueira ◽  
Nuno C. Santos
Keyword(s):  
Cell Adhesion ◽  
Cardiovascular Risk ◽  
Arterial Hypertension ◽  
Adhesion Forces ◽  
Essential Arterial Hypertension

Arterial hypertension patients present stronger erythrocyte–erythrocyte interactions and high levels of γ′ fibrinogen, which compromise microcirculation and increase the cardiovascular risk.

scholarly journals Methods for Voltage-Sensitive Dye Imaging of Rat Cortical Activity With High Signal-to-Noise Ratio

2007 ◽  
Vol 98 (1) ◽  
pp. 502-512 ◽  
Author(s):  
Michael T. Lippert ◽  
Kentaroh Takagaki ◽  
Weifeng Xu ◽  
Xiaoying Huang ◽  
Jian-Young Wu
Keyword(s):  
Dynamic Range ◽  
Signal To Noise Ratio ◽  
Field Potential ◽  
High Sensitivity ◽  
High Dynamic Range ◽  
High Signal ◽  
Blue Dye ◽  
Voltage Sensitive Dye ◽  
Imaging Device

We describe methods to achieve high sensitivity in voltage-sensitive dye (VSD) imaging from rat barrel and visual cortices in vivo with the use of a blue dye RH1691 and a high dynamic range imaging device (photodiode array). With an improved staining protocol and an off-line procedure to remove pulsation artifact, the sensitivity of VSD recording is comparable with that of local field potential recording from the same location. With this sensitivity, one can record from ∼500 individual detectors, each covering an area of cortical tissue 160 μm in diameter (total imaging field ∼4 mm in diameter) and a temporal resolution of 1,600 frames/s, without multiple-trial averaging. We can record 80–100 trials of intermittent 10-s trials from each imaging field before the VSD signal reduces to one half of its initial amplitude because of bleaching and wash-out. Taken together, the methods described in this report provide a useful tool for visualizing evoked and spontaneous waves from rodent cortex.

scholarly journals Quantification of fast molecular adhesion by fluorescence footprinting

2021 ◽  
Author(s):  
Adam B. Yasunaga ◽  
Isaac T.S. Li
Keyword(s):  
Shear Stress ◽  
Dynamic Range ◽  
Force Distribution ◽  
Equilibrium Conditions ◽  
Ligand Dissociation ◽  
Molecular Adhesion ◽  
Molecular Force ◽  
Adhesion Complex ◽  
Non Equilibrium

AbstractRolling adhesion is a unique process in which the adhesion events are short-lived and operate under highly non-equilibrium conditions. These characteristics pose a challenge in molecular force quantification, where in situ measurement of such forces cannot be achieved with most molecular force sensors that probe near equilibrium. In this report, we demonstrated a quantitative adhesion footprint assay combining DNA-based non-equilibrium force probes and modelling to measure the molecular force involved in fast rolling adhesion. We were able to directly profile the ensemble molecular force distribution during rolling adhesion with a dynamic range between 0 – 18 pN. Our results showed that the shear stress driving bead rolling motility directly controls the molecular tension on the probe-conjugated adhesion complex. Furthermore, the shear stress can steer the dissociation bias of components within the molecular force probe complex, favouring either DNA probe dissociation or receptor-ligand dissociation.

scholarly journals Novel Electrochemiluminescent Assays for Drug Discovery

2000 ◽  
Vol 5 (1) ◽  
pp. 45-48 ◽  
Author(s):  
Maura C. Kibbey ◽  
David MacAllan ◽  
James W. Karaszkiewicz
Keyword(s):  
Drug Discovery ◽  
High Precision ◽  
High Throughput ◽  
High Performance ◽  
Dynamic Range ◽  
Wide Dynamic Range ◽  
Receptor Ligand ◽  
Enzymatic Assays ◽  
Detection Systems ◽  
Protein Assays

IGEN's ORIGEN® technology, which is based on electrochemiluminescence, has been adopted by a number of research and bioanalytical laboratories who have recognized its exquisite sensitivity, high precision, wide dynamic range, and flexibility in formatting a wide variety of applications. IGEN's M-SERIES™ marks the introduction of the second generation of detection systems employing the ORIGEN technology specifically repackaged to address the needs of the high throughput laboratories involved in drug discovery. Assays are formatted without wash steps. Users realize the high performance of a heterogeneous technology with the convenience of a homogeneous format. The M-SERIES platform can address enzymatic assays (kinases, proteases, helicases, etc.), receptor-ligand or protein-protein assays, immunoassays, quantitation of nucleic acids, as well as other applications. Recent assay formats will be explored in detail.

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