In the last two decades the study of red blood cell elasticity using optical tweezers has known a rise appearing in the scientific research with regard to the various works carried out. Despite the various work done, ...In the last two decades the study of red blood cell elasticity using optical tweezers has known a rise appearing in the scientific research with regard to the various works carried out. Despite the various work done, no study has been done so far to study the influence of friction on the red blood cell indentation response using optical tweezers. In this study, we have developed a new approach to determine the coefficient of friction as well as the frictional forces of the red blood cell. This approach therefore allowed us to simultaneously carry out the indentation and traction test, which allowed us to extract the interfacial properties of the microbead red blood cell couple, among other things, the friction coefficient. This property would be extremely important to investigate the survival and mechanical features of cells, which will be of great physiological and pathological significance. But taking into account the hypothesis of friction as defined by the isotropic Coulomb law. The experiment performed for this purpose is the Brinell Hardness Test (DB).展开更多
With the advantages of noncontact,high accuracy,and high flexibility,optical tweezers hold huge potential for micro-manipulation and force measurement.However,the majority of previous research focused on the state of ...With the advantages of noncontact,high accuracy,and high flexibility,optical tweezers hold huge potential for micro-manipulation and force measurement.However,the majority of previous research focused on the state of the motion of particles in the optical trap,but paid little attention to the early dynamic process between the initial state of the particles and the optical trap.Note that the viscous forces can greatly affect the motion of micro-spheres.In this paper,based on the equations of Newtonian mechanics,we investigate the dynamics of laser-trapped micro-spheres in the surrounding environment with different viscosity coefficients.Through the calculations,over time the particle trajectory clearly reveals the subtle details of the optical capture process,including acceleration,deceleration,turning,and reciprocating oscillation.The time to equilibrium mainly depends on the corresponding damping coefficient of the surrounding environment and the oscillation frequency of the optical tweezers.These studies are essential for understanding various mechanisms to engineer the mechanical motion behavior of molecules or microparticles in liquid or air.展开更多
A facile,rapid and efficient method for the synthesis of molecular tweezers containing chiral unsymmetrical urea unit in solvent- free conditions using microwave was reported.
The well understanding of interaction forces between single dispersed droplets is crucial to the understanding of emulsion stabilization mechanism.Recently,many studies have reported the direct quantitative measuremen...The well understanding of interaction forces between single dispersed droplets is crucial to the understanding of emulsion stabilization mechanism.Recently,many studies have reported the direct quantitative measurements of interaction forces between 20-200μm single droplet coated polymers by atomic force microscope(AFM).These studies have revealed many important results about the relationship of the interaction forces and the droplet deformation.However,these studies of the quantitative relationship between the measured interaction forces and the separation distance of the front end of the droplet have rarely been reported.Optical tweezer instrument can make it possible to establish the quantitative relationship between the measured force and the separation distance of the front end of the droplet,which will make better understanding of the interaction mechanisms between droplets.Due to the differences of the measuring mechanism between atomic force microscopy(AFM)and optical tweezers,the theory model of AFM measurements cannot be fitted with the force measurement by optical tweezers.We have made an exhaustive comparison of the measuring differences between AFM and optical tweezer instrument in this work.Moreover,we built a numerical model to derive the repulsive pressure through the measured force curve in order to quantify the measured force of two micron-sized oil droplet coated polymers by optical tweezers.Furthermore,the novel method can be extended to other micron-sized emulsion systems,and these findings will be a vital progress on quantitative force measurements between micron-sized droplets.展开更多
The rotation control of particles in optical tweezers is often subject to the spin or orbit angular momentum induced optical torque,which is susceptible to the mechanical and morphological properties of individual par...The rotation control of particles in optical tweezers is often subject to the spin or orbit angular momentum induced optical torque,which is susceptible to the mechanical and morphological properties of individual particle.Here we report on a robust and high-speed rotation control in optical tweezers by using a novel linear polarization synthesis based on optical heterodyne interference between two circularly polarized lights with opposite handedness.The synthesized linear polarization can be rotated in a hopping-free scheme at arbitrary speed determined electronically by the heterodyne frequency between two laser fields.The experimental demonstration of a trapped vaterite particle in water shows that the precisely controlled rotation frequency of 300 Hz can be achieved.The proposed method will find promising applications in optically driven micro-gears,fluidic pumps and rotational micro-rheology.展开更多
Src SH3 protein domain is a typical two-state protein which has been confirmed by research of denaturant-induced unfolding dynamics.Force spectroscopy experiments by optical tweezers and atomic force microscopy have m...Src SH3 protein domain is a typical two-state protein which has been confirmed by research of denaturant-induced unfolding dynamics.Force spectroscopy experiments by optical tweezers and atomic force microscopy have measured the force-dependent unfolding rates with different kinds of pulling geometry.However,the equilibrium folding and unfolding dynamics at constant forces has not been reported.Here,using stable magnetic tweezers,we performed equilibrium folding and unfolding dynamic measurement and force-jump measurement of src SH3 domain with tethering points at its N-and C-termini.From the obtained force-dependent transition rates,a detailed two-state free energy landscape of src SH3 protein is constructed with quantitative information of folding free energy,transition state barrier height and position,which exemplifies the capability of magnetic tweezers to study protein folding and unfolding dynamics.展开更多
As studies on life sciences progress toward the single-molecule level,new experiments have put forward more requirements for simultaneously displaying the mechanical properties and conformational changes of biomolecul...As studies on life sciences progress toward the single-molecule level,new experiments have put forward more requirements for simultaneously displaying the mechanical properties and conformational changes of biomolecules.Optical tweezers and fluorescence microscopy have been combined to solve this problem.The combination of instruments forms a new generation of hybrid single-molecule technology that breaks through the limitations of traditional biochemical analysis.Powerfulmanipulation and fluorescence visualization have beenwidely used,and these techniques provide new possibilities for studying complex biochemical reactions at the singlemolecule level.This paper explains the features of this combined technique,including the application characteristics of single-trap and dual-traps,the anti-bleaching method,and optical tweezers combined with epifluorescence,confocal fluorescence,total internal reflection fluorescence,and other fluorescence methods.Using typical experiments,we analyze technical solutions and explain the factors and principles that instrument designers should consider.This review aims to give an introduction to this novel fusion technology process and describe important biological results.展开更多
Known as laser trapping,optical tweezers,with nanometer accuracy and pico-newton precision,plays a pivotal role in single bio-molecule measurements and controllable motions of micro-machines.In order to advance the fl...Known as laser trapping,optical tweezers,with nanometer accuracy and pico-newton precision,plays a pivotal role in single bio-molecule measurements and controllable motions of micro-machines.In order to advance the flourishing applications for those achievements,it is necessary to make clear the three-dimensional dynamic process of micro-particles stepping into an optical field.In this paper,we utilize the ray optics method to calculate the optical force and optical torque of a micro-sphere in optical tweezers.With the influence of viscosity force and torque taken into account,we numerically solve and analyze the dynamic process of a dielectric micro-sphere in optical tweezers on the basis of Newton mechanical equations under various conditions of initial positions and velocity vectors of the particle.The particle trajectory over time can demonstrate whether the particle can be successfully trapped into the optical tweezers center and reveal the subtle details of this trapping process.Even in a simple pair of optical tweezers,the dielectric micro-sphere exhibits abundant phases of mechanical motions including acceleration,deceleration,and turning.These studies will be of great help to understand the particle-laser trap interaction in various situations and promote exciting possibilities for exploring novel ways to control the mechanical dynamics of microscale particles.展开更多
Contactless acoustic manipulation of micro/nanoscale particles has attracted considerable attention owing to its near independence of the physical and chemical properties of the targets,making it universally applicabl...Contactless acoustic manipulation of micro/nanoscale particles has attracted considerable attention owing to its near independence of the physical and chemical properties of the targets,making it universally applicable to almost all biological systems.Thin-film bulk acoustic wave(BAW)resonators operating at gigahertz(GHz)frequencies have been demonstrated to generate localized high-speed microvortices through acoustic streaming effects.Benefitting from the strong drag forces of the high-speed vortices,BAW-enabled GHz acoustic streaming tweezers(AST)have been applied to the trapping and enrichment of particles ranging in size from micrometers to less than 100 nm.However,the behavior of particles in such 3D microvortex systems is still largely unknown.In this work,the particle behavior(trapping,enrichment,and separation)in GHz AST is studied by theoretical analyses,3D simulations,and microparticle tracking experiments.It is found that the particle motion in the vortices is determined mainly by the balance between the acoustic streaming drag force and the acoustic radiation force.This work can provide basic design principles for AST-based lab-on-a-chip systems for a variety of applications.展开更多
Cells actively modulate mechanobiological circuitry against external perturbations to stabilize whole cell/tissue physiology.The dynamic adaption of cells to mechanical force is critical for cells to perform vital bio...Cells actively modulate mechanobiological circuitry against external perturbations to stabilize whole cell/tissue physiology.The dynamic adaption of cells to mechanical force is critical for cells to perform vital biological functions,from single cell migration to embryonic development.Dysregulation of such dynamics has been associated with pathophysiological conditions in cardiovascular diseases,cancer,aging,and developmental disorders[1].Therefore,a direct understanding of cell’s biomechanical adaptive/maladaptive behaviors and the trigger factors causing the transformation of healthy adaption to maladaptation can help reveal the regulatory role of single cell mechanosensitive dynamics in the progression of various degenerative diseases and aging.However,current efforts for uncovering fundamental associations between disease and cell architecture have been focusing on'static'measurements of biophysical properties,which is limited by the requirement of large sample sizes to obtain statistically significant data.We therefore developed a single and highly integrated platform with mechanical stimulation and fine spatiotemporal sensing functions to probe the single cell mechanical dynamics at subcellular level to determine cell’s mechanophenotypes in healthy and disease conditions.We developed an integrated micromechanical system composed of an’ultrasound tweezer’stimulator[2]and a PDMS micropillar array [3] cellular force sensor to in situ noninvasively probe and monitor single cell mechanical dynamics.Vascular smooth muscle cells(VSMCs)from healthy mouse and mouse with induced abdominal aorta aneurysm(AAA)were used for cell mechanobiological study.An ultrasound transducer(V312-SM,Olympus)was used to generate ultrasound pulses to excite lipid-encapsulated microbubbles(Targeson)binding to cell membrane through an RGD-integrin linkage to apply a transient nanonewton force to VSMCs seeded on the PDMS micropillar array.PDMS micropillar array was fabricated and functionalized as previously described [3] and acts as the mechanical force sensor in our platform.Upon a 1 HZ and 10-second ultrasound stimulation,calcium influx was clearly detected in both healthy and AAA-VSMCs by using the fluo-4 calcium sensor,suggesting the microbubble-integrin-actin cytoskeleton(CSK)linkage can serve as a mechanosensory to sense the ultrasound stimulation.We then examined how healthy and AAA VSMCs would exhibit adaptions to mechanical stimulation at a global cellular scale.After the onset of a 10-second ultrasound stimulation,control and AAA-VSMCs displayed distinct dynamics of CSK tension within 30 mins,in which the CSK tension of healthy VSMCs increased within the reinforcement period(0-5 min)and restored to their ground state with the relaxation period(5-10 min);yet AAA-VSMCs displayed compromised dynamics of such CSK tension upon calcium influx.Quantitative analysis and theoretical modelling revealed the critical roles of myosin motor contraction,F-actin filament polymerization in regulating cell mechanosensitive dynamics in response to a transient mechanical perturbation.The distinct force and CSK dynamics in healthy and AAA conditions indicates that the force-dependent CSK molecular kinetics is a critical factor governing the distinct mechanosensitive dynamics of cells under pathologically dysfunctional conditions.Our results reveal that the mechanical adaptive process of cells to mechanical stimulus can measure the cellular mechanobiological phenotypes featured in both pathologically healthy and diseased context.We demonstrated that an altered mechanobiological phenotype,i.e.AAA-VSMCs with distinct actomyosin-CSK properties potentiates a mechanical maladaptation that reflects progressive accumulation of cellular damage and dysfunction.This may further reveal the pathogenic contexts and their physical mediators featuring biophysical dysregulation in cardiovascular diseases.展开更多
A series of new chiral molecular tweezers have been designed and synthesized by using deoxycholic acid as spacer and aromatic amines as arms. Instead of using toxic phosgene, the triphosgene was employed in synthesis ...A series of new chiral molecular tweezers have been designed and synthesized by using deoxycholic acid as spacer and aromatic amines as arms. Instead of using toxic phosgene, the triphosgene was employed in synthesis of the molecular tweezers receptors. These chiral molecular tweezers showed good enantioselectivity for D-amino acid methyl esters.展开更多
Mammalian cell surfaces consist of the plasma membrane supported by an underneath cortical cytoskeleton.Together,these structures can control not only the shape of cells but also a series of cellular functions ranging...Mammalian cell surfaces consist of the plasma membrane supported by an underneath cortical cytoskeleton.Together,these structures can control not only the shape of cells but also a series of cellular functions ranging from migration and division to exocytosis,endocytosis and differentiation.Furthermore,the cell surface is capable of exerting and reacting to mechanical forces.Its viscoelastic properties,especially membrane tension and bending modulus,are fundamental parameters involved in these responses.This viewpoint summarizes our current knowledge on how to measure the viscoelastic properties of cell surfaces employing optical tweezers-based tether assays,paving the way for a better understanding of how cells react to external mechanical forces,with a glance on their remodeling dynamics and possible consequences on downstream cellular processes.展开更多
In recent years, extensive research has been carried out on red blood cells in order to investigate their mechanical properties. The interest in these studies has been possible thanks to the technological innovations ...In recent years, extensive research has been carried out on red blood cells in order to investigate their mechanical properties. The interest in these studies has been possible thanks to the technological innovations made in the field of micro or nano manipulation of biological and non-biological particles without physical contact. In the present project, we have developed a new approach to study the deformation of red blood cells moving against a trapped microbead by applying a sinusoidal voltage (DC offset 3.5 Vpp) to the stage at 0.4 Hz frequencies. The oscillating movement imposed on the stage highlights the indentation test and the tensile test known for the study of mechanical behavior of materials. The mechanical properties found are: the modulus of elasticity (Young Modulus), the shear modulus, the coefficient of hardening and erythrocyte resistance coefficient. The axial shear modulus 25.00 ± 1.5 μN/m and the transversal shear modulus 15.7 ± 4.63 μN/m were compared to those in the literature. These values were respectively determined by Hooke’s law and the Hertz model.展开更多
Several attempts have appeared recently to control optical trapping systems via touch tablets and cameras instead of a mouse and joystick. Our approach is based on a modern low-cost hardware combined with fingertips a...Several attempts have appeared recently to control optical trapping systems via touch tablets and cameras instead of a mouse and joystick. Our approach is based on a modern low-cost hardware combined with fingertips and speech recognition software. Positions of operator's hands or fingertips control the positions of trapping beams in holographic optical tweezers that provide optical manipulation with microobjects. We tested and adapted two systems for hands position detection and gestures recognition – Creative Interactive Gesture Camera and Leap Motion. We further enhanced the system of Holographic Raman tweezers (HRT) by voice commands controlling the micropositioning stage and acquisition of Raman spectra. Interface communicates with HRT either directly by which requires adaptation of HRT firmware, or indirectly by simulating mouse and keyboard messages. Its utilization in real experiments speeded up the operator’s communication with the system cca. Two times in comparison with the traditional control by the mouse and the keyboard.展开更多
Optical tweezers that rely on laser irradiation to capture and manipulate nanoparticles have provided powerful tools for biological and biochemistry studies.However,the existence of optical diffraction-limit and the t...Optical tweezers that rely on laser irradiation to capture and manipulate nanoparticles have provided powerful tools for biological and biochemistry studies.However,the existence of optical diffraction-limit and the thermal damage caused by high laser power hinder the wider application of optical tweezers in the biological field.For the past decade,the emergence of optothermal tweezers has solved the above problems to a certain extent,while the auxiliary agents used in optothermal tweezers still limit their biocompatibility.Here,we report a kind of nanotweezers based on the sign transformation of the thermophoresis coefficient of colloidal particles in low-temperature environment.Using a self-made microfluidic refrigerator to reduce the ambient temperature to around 0℃in the microfluidic cell,we can control a single nanoparticle at lower laser power without adding additional agent solute in the solution.This novel optical tweezering scheme has provided a new path for the manipulation of inorganic nanoparticles as well as biological particles.展开更多
Sensing sensitivity is the key performance of optical tweezers.By adjusting the frequency and magnitude of an applied Coulomb force as an input of optical tweezers,we directly measured the sensitivity and signal-to-no...Sensing sensitivity is the key performance of optical tweezers.By adjusting the frequency and magnitude of an applied Coulomb force as an input of optical tweezers,we directly measured the sensitivity and signal-to-noise ratio(SNR)of a system and indirectly calculated the actual noise magnitude.Combined with an output filter,the relationship between the SNR and bandwidths was studied.We established the simulation model of a system using Simulink and simulated the relationship between the SNR and magnitude of the input forces and filter bandwidths.In addition,we built an experimental system to determine the relationship between the SNR and the magnitude of the input forces and filter bandwidths.The actual minimum detectable force was measured as 1.8275×10^(-17)N at a 1Hz bandwidth.The experimental results were correlated with the simulation and theoretical results,confirming the effectiveness of the proposed method and demonstrating the high sensitivity of vacuum optical tweezers as mechanical sensors.We proposed a novel method of calibration and measurement of system sensing parameters by applying an actual force that was more direct and precise than the theoretical calculation method that requires accurate fitting parameters,such as the particle radius and density.This method can be employed to analyze the system noise and phase characteristics to confirm and improve the real performance of the system.展开更多
In recent years,levitated particles of optical traps in vacuum have shown the enormous potential for precision sensor development and new physics exploration.However,the accuracy of the sensor is still hampered by the...In recent years,levitated particles of optical traps in vacuum have shown the enormous potential for precision sensor development and new physics exploration.However,the accuracy of the sensor is still hampered by the uncertainty of the calibration factor relating the detected signal to the absolute displacement of the trapped particle.In this paper,we suggest and experimentally demonstrate a novel calibration method for optical tweezers based on free-falling particles in vacuum,where the gravitational acceleration is introduced as an absolute reference.Our work provides a calibration protocol with a great certainty and traceability,which is significant in improving the accuracy of precision sensing based on levitated optomechanical systems.展开更多
Acoustic radiation force(ARF), as an important particle manipulation method, has been extensively studied in recent years. With the introduction of the concept of “acoustic tweezers”, negative acoustic radiation has...Acoustic radiation force(ARF), as an important particle manipulation method, has been extensively studied in recent years. With the introduction of the concept of “acoustic tweezers”, negative acoustic radiation has become a research hotspot. In this paper, a scheme of realizing negative ARF based on the multiple-layered spherical structure design is proposed. The specific structure and design idea are presented. Detailed theoretical calculation analysis is carried out.Numerical simulations have been performed to verify the correctness of this prediction. The conjecture that the suppression of backscattering can achieve negative ARF is verified concretely, which greatly expands the application prospect and design ideas of the ARF. This work has laid a theoretical foundation for realizing precise control of the structure.展开更多
Optical manipulation of micro/nanoscale objects is of importance in life sciences,colloidal science,and nanotechnology.Optothermal tweezers exhibit superior manipulation capability at low optical intensity.However,our...Optical manipulation of micro/nanoscale objects is of importance in life sciences,colloidal science,and nanotechnology.Optothermal tweezers exhibit superior manipulation capability at low optical intensity.However,our implicit understanding of the working mechanism has limited the further applications and innovations of optothermal tweezers.Herein,we present an atomistic view of opto-thermo-electro-mechanic coupling in optothermal tweezers,which enables us to rationally design the tweezers for optimum performance in targeted applications.Specifically,we have revealed that the non-uniform temperature distribution induces water polarization and charge separation,which creates the thermoelectric field dominating the optothermal trapping.We further design experiments to systematically verify our atomistic simulations.Guided by our new model,we develop new types of optothermal tweezers of high performance using low-concentrated electrolytes.Moreover,we demonstrate the use of new tweezers in opto-thermophoretic separation of colloidal particles of the same size based on the difference in their surface charge,which has been challenging for conventional optical tweezers.With the atomistic understanding that enables the performance optimization and function expansion,optothermal tweezers will further their impacts.展开更多
文摘In the last two decades the study of red blood cell elasticity using optical tweezers has known a rise appearing in the scientific research with regard to the various works carried out. Despite the various work done, no study has been done so far to study the influence of friction on the red blood cell indentation response using optical tweezers. In this study, we have developed a new approach to determine the coefficient of friction as well as the frictional forces of the red blood cell. This approach therefore allowed us to simultaneously carry out the indentation and traction test, which allowed us to extract the interfacial properties of the microbead red blood cell couple, among other things, the friction coefficient. This property would be extremely important to investigate the survival and mechanical features of cells, which will be of great physiological and pathological significance. But taking into account the hypothesis of friction as defined by the isotropic Coulomb law. The experiment performed for this purpose is the Brinell Hardness Test (DB).
基金Project supported by the National Natural Science Foundation of China(Grant No.11804399)the Special Funds for Basic Scientific Research at the Central University of South-Central University for Nationalities(Grant No.CZQ20018)Special Funds for Basic Scientific Research at Central Universities(Grant No.YZZ17005)。
文摘With the advantages of noncontact,high accuracy,and high flexibility,optical tweezers hold huge potential for micro-manipulation and force measurement.However,the majority of previous research focused on the state of the motion of particles in the optical trap,but paid little attention to the early dynamic process between the initial state of the particles and the optical trap.Note that the viscous forces can greatly affect the motion of micro-spheres.In this paper,based on the equations of Newtonian mechanics,we investigate the dynamics of laser-trapped micro-spheres in the surrounding environment with different viscosity coefficients.Through the calculations,over time the particle trajectory clearly reveals the subtle details of the optical capture process,including acceleration,deceleration,turning,and reciprocating oscillation.The time to equilibrium mainly depends on the corresponding damping coefficient of the surrounding environment and the oscillation frequency of the optical tweezers.These studies are essential for understanding various mechanisms to engineer the mechanical motion behavior of molecules or microparticles in liquid or air.
文摘A facile,rapid and efficient method for the synthesis of molecular tweezers containing chiral unsymmetrical urea unit in solvent- free conditions using microwave was reported.
基金the supports of the National Natural Science Foundation of China(2147612121322604)for this work。
文摘The well understanding of interaction forces between single dispersed droplets is crucial to the understanding of emulsion stabilization mechanism.Recently,many studies have reported the direct quantitative measurements of interaction forces between 20-200μm single droplet coated polymers by atomic force microscope(AFM).These studies have revealed many important results about the relationship of the interaction forces and the droplet deformation.However,these studies of the quantitative relationship between the measured interaction forces and the separation distance of the front end of the droplet have rarely been reported.Optical tweezer instrument can make it possible to establish the quantitative relationship between the measured force and the separation distance of the front end of the droplet,which will make better understanding of the interaction mechanisms between droplets.Due to the differences of the measuring mechanism between atomic force microscopy(AFM)and optical tweezers,the theory model of AFM measurements cannot be fitted with the force measurement by optical tweezers.We have made an exhaustive comparison of the measuring differences between AFM and optical tweezer instrument in this work.Moreover,we built a numerical model to derive the repulsive pressure through the measured force curve in order to quantify the measured force of two micron-sized oil droplet coated polymers by optical tweezers.Furthermore,the novel method can be extended to other micron-sized emulsion systems,and these findings will be a vital progress on quantitative force measurements between micron-sized droplets.
基金the National Natural Science Foundation of China(91750203 and 91850111)State Key Laboratory of Applied Optics,Changchun Institute of Optics,Fine Mechanics and Physics,Chinese Academy of Sciences and the High-performance Computing Platform of Peking University.
文摘The rotation control of particles in optical tweezers is often subject to the spin or orbit angular momentum induced optical torque,which is susceptible to the mechanical and morphological properties of individual particle.Here we report on a robust and high-speed rotation control in optical tweezers by using a novel linear polarization synthesis based on optical heterodyne interference between two circularly polarized lights with opposite handedness.The synthesized linear polarization can be rotated in a hopping-free scheme at arbitrary speed determined electronically by the heterodyne frequency between two laser fields.The experimental demonstration of a trapped vaterite particle in water shows that the precisely controlled rotation frequency of 300 Hz can be achieved.The proposed method will find promising applications in optically driven micro-gears,fluidic pumps and rotational micro-rheology.
基金the National Natural Science Foundation of China(Grant Nos.11874309 and 11474237)the 111 Project(Grant No.B16029)。
文摘Src SH3 protein domain is a typical two-state protein which has been confirmed by research of denaturant-induced unfolding dynamics.Force spectroscopy experiments by optical tweezers and atomic force microscopy have measured the force-dependent unfolding rates with different kinds of pulling geometry.However,the equilibrium folding and unfolding dynamics at constant forces has not been reported.Here,using stable magnetic tweezers,we performed equilibrium folding and unfolding dynamic measurement and force-jump measurement of src SH3 domain with tethering points at its N-and C-termini.From the obtained force-dependent transition rates,a detailed two-state free energy landscape of src SH3 protein is constructed with quantitative information of folding free energy,transition state barrier height and position,which exemplifies the capability of magnetic tweezers to study protein folding and unfolding dynamics.
基金supported by the National Key Research and Development Program of China [grant numbers 2016YFB1102203, 2017YFF0107003]
文摘As studies on life sciences progress toward the single-molecule level,new experiments have put forward more requirements for simultaneously displaying the mechanical properties and conformational changes of biomolecules.Optical tweezers and fluorescence microscopy have been combined to solve this problem.The combination of instruments forms a new generation of hybrid single-molecule technology that breaks through the limitations of traditional biochemical analysis.Powerfulmanipulation and fluorescence visualization have beenwidely used,and these techniques provide new possibilities for studying complex biochemical reactions at the singlemolecule level.This paper explains the features of this combined technique,including the application characteristics of single-trap and dual-traps,the anti-bleaching method,and optical tweezers combined with epifluorescence,confocal fluorescence,total internal reflection fluorescence,and other fluorescence methods.Using typical experiments,we analyze technical solutions and explain the factors and principles that instrument designers should consider.This review aims to give an introduction to this novel fusion technology process and describe important biological results.
基金This work is supported by the National Natural Science Foundation of China(Grant No.11974119 and No.11804399)the Guangdong Innovative and Entrepreneurial Research Team Program(Grant No.2016ZT06C594)+1 种基金the Fundamental Research Funds for the Central Universities,South-Central University for Nationalities(Grant No.CZQ20018)National Key R&D Program of China(No.2018YFA 0306200).
文摘Known as laser trapping,optical tweezers,with nanometer accuracy and pico-newton precision,plays a pivotal role in single bio-molecule measurements and controllable motions of micro-machines.In order to advance the flourishing applications for those achievements,it is necessary to make clear the three-dimensional dynamic process of micro-particles stepping into an optical field.In this paper,we utilize the ray optics method to calculate the optical force and optical torque of a micro-sphere in optical tweezers.With the influence of viscosity force and torque taken into account,we numerically solve and analyze the dynamic process of a dielectric micro-sphere in optical tweezers on the basis of Newton mechanical equations under various conditions of initial positions and velocity vectors of the particle.The particle trajectory over time can demonstrate whether the particle can be successfully trapped into the optical tweezers center and reveal the subtle details of this trapping process.Even in a simple pair of optical tweezers,the dielectric micro-sphere exhibits abundant phases of mechanical motions including acceleration,deceleration,and turning.These studies will be of great help to understand the particle-laser trap interaction in various situations and promote exciting possibilities for exploring novel ways to control the mechanical dynamics of microscale particles.
基金The authors gratefully acknowledge financial support from the National Key R&D Program of China(2018YFE0118700)the Natural Science Foundation of China(NSFC No.62174119)+1 种基金Tianjin Applied Basic Research and Advanced Technology(17JCJQJC43600)the 111 Project(B07014).
文摘Contactless acoustic manipulation of micro/nanoscale particles has attracted considerable attention owing to its near independence of the physical and chemical properties of the targets,making it universally applicable to almost all biological systems.Thin-film bulk acoustic wave(BAW)resonators operating at gigahertz(GHz)frequencies have been demonstrated to generate localized high-speed microvortices through acoustic streaming effects.Benefitting from the strong drag forces of the high-speed vortices,BAW-enabled GHz acoustic streaming tweezers(AST)have been applied to the trapping and enrichment of particles ranging in size from micrometers to less than 100 nm.However,the behavior of particles in such 3D microvortex systems is still largely unknown.In this work,the particle behavior(trapping,enrichment,and separation)in GHz AST is studied by theoretical analyses,3D simulations,and microparticle tracking experiments.It is found that the particle motion in the vortices is determined mainly by the balance between the acoustic streaming drag force and the acoustic radiation force.This work can provide basic design principles for AST-based lab-on-a-chip systems for a variety of applications.
基金the financial support from the American Heart Association ( 16SDG31020038)
文摘Cells actively modulate mechanobiological circuitry against external perturbations to stabilize whole cell/tissue physiology.The dynamic adaption of cells to mechanical force is critical for cells to perform vital biological functions,from single cell migration to embryonic development.Dysregulation of such dynamics has been associated with pathophysiological conditions in cardiovascular diseases,cancer,aging,and developmental disorders[1].Therefore,a direct understanding of cell’s biomechanical adaptive/maladaptive behaviors and the trigger factors causing the transformation of healthy adaption to maladaptation can help reveal the regulatory role of single cell mechanosensitive dynamics in the progression of various degenerative diseases and aging.However,current efforts for uncovering fundamental associations between disease and cell architecture have been focusing on'static'measurements of biophysical properties,which is limited by the requirement of large sample sizes to obtain statistically significant data.We therefore developed a single and highly integrated platform with mechanical stimulation and fine spatiotemporal sensing functions to probe the single cell mechanical dynamics at subcellular level to determine cell’s mechanophenotypes in healthy and disease conditions.We developed an integrated micromechanical system composed of an’ultrasound tweezer’stimulator[2]and a PDMS micropillar array [3] cellular force sensor to in situ noninvasively probe and monitor single cell mechanical dynamics.Vascular smooth muscle cells(VSMCs)from healthy mouse and mouse with induced abdominal aorta aneurysm(AAA)were used for cell mechanobiological study.An ultrasound transducer(V312-SM,Olympus)was used to generate ultrasound pulses to excite lipid-encapsulated microbubbles(Targeson)binding to cell membrane through an RGD-integrin linkage to apply a transient nanonewton force to VSMCs seeded on the PDMS micropillar array.PDMS micropillar array was fabricated and functionalized as previously described [3] and acts as the mechanical force sensor in our platform.Upon a 1 HZ and 10-second ultrasound stimulation,calcium influx was clearly detected in both healthy and AAA-VSMCs by using the fluo-4 calcium sensor,suggesting the microbubble-integrin-actin cytoskeleton(CSK)linkage can serve as a mechanosensory to sense the ultrasound stimulation.We then examined how healthy and AAA VSMCs would exhibit adaptions to mechanical stimulation at a global cellular scale.After the onset of a 10-second ultrasound stimulation,control and AAA-VSMCs displayed distinct dynamics of CSK tension within 30 mins,in which the CSK tension of healthy VSMCs increased within the reinforcement period(0-5 min)and restored to their ground state with the relaxation period(5-10 min);yet AAA-VSMCs displayed compromised dynamics of such CSK tension upon calcium influx.Quantitative analysis and theoretical modelling revealed the critical roles of myosin motor contraction,F-actin filament polymerization in regulating cell mechanosensitive dynamics in response to a transient mechanical perturbation.The distinct force and CSK dynamics in healthy and AAA conditions indicates that the force-dependent CSK molecular kinetics is a critical factor governing the distinct mechanosensitive dynamics of cells under pathologically dysfunctional conditions.Our results reveal that the mechanical adaptive process of cells to mechanical stimulus can measure the cellular mechanobiological phenotypes featured in both pathologically healthy and diseased context.We demonstrated that an altered mechanobiological phenotype,i.e.AAA-VSMCs with distinct actomyosin-CSK properties potentiates a mechanical maladaptation that reflects progressive accumulation of cellular damage and dysfunction.This may further reveal the pathogenic contexts and their physical mediators featuring biophysical dysregulation in cardiovascular diseases.
基金We are very grateful to the National Natural Science Foundation of China(project:No.29772024)for the financial support.
文摘A series of new chiral molecular tweezers have been designed and synthesized by using deoxycholic acid as spacer and aromatic amines as arms. Instead of using toxic phosgene, the triphosgene was employed in synthesis of the molecular tweezers receptors. These chiral molecular tweezers showed good enantioselectivity for D-amino acid methyl esters.
基金This work was supported by the Brazilian agencies Conselho Nacional de Desenvolvimento Científico e Tecnológico(CNPq)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior(CAPES)–Financial Code 001,Fundação de AmparoàPesquisa do Estado do Rio de Janeiro(FAPERJ)+1 种基金Fundação de AmparoàPesquisa do Estado do Amazonas(FAPEAM)Instituto Nacional de Ciência e Tecnologia de Fluidos Complexos(INCT-FCx)together with Fundação de AmparoàPesquisa do Estado de São Paulo(FAPESP).BP was supported by a JCNE grant from FAPERJ.
文摘Mammalian cell surfaces consist of the plasma membrane supported by an underneath cortical cytoskeleton.Together,these structures can control not only the shape of cells but also a series of cellular functions ranging from migration and division to exocytosis,endocytosis and differentiation.Furthermore,the cell surface is capable of exerting and reacting to mechanical forces.Its viscoelastic properties,especially membrane tension and bending modulus,are fundamental parameters involved in these responses.This viewpoint summarizes our current knowledge on how to measure the viscoelastic properties of cell surfaces employing optical tweezers-based tether assays,paving the way for a better understanding of how cells react to external mechanical forces,with a glance on their remodeling dynamics and possible consequences on downstream cellular processes.
文摘In recent years, extensive research has been carried out on red blood cells in order to investigate their mechanical properties. The interest in these studies has been possible thanks to the technological innovations made in the field of micro or nano manipulation of biological and non-biological particles without physical contact. In the present project, we have developed a new approach to study the deformation of red blood cells moving against a trapped microbead by applying a sinusoidal voltage (DC offset 3.5 Vpp) to the stage at 0.4 Hz frequencies. The oscillating movement imposed on the stage highlights the indentation test and the tensile test known for the study of mechanical behavior of materials. The mechanical properties found are: the modulus of elasticity (Young Modulus), the shear modulus, the coefficient of hardening and erythrocyte resistance coefficient. The axial shear modulus 25.00 ± 1.5 μN/m and the transversal shear modulus 15.7 ± 4.63 μN/m were compared to those in the literature. These values were respectively determined by Hooke’s law and the Hertz model.
文摘Several attempts have appeared recently to control optical trapping systems via touch tablets and cameras instead of a mouse and joystick. Our approach is based on a modern low-cost hardware combined with fingertips and speech recognition software. Positions of operator's hands or fingertips control the positions of trapping beams in holographic optical tweezers that provide optical manipulation with microobjects. We tested and adapted two systems for hands position detection and gestures recognition – Creative Interactive Gesture Camera and Leap Motion. We further enhanced the system of Holographic Raman tweezers (HRT) by voice commands controlling the micropositioning stage and acquisition of Raman spectra. Interface communicates with HRT either directly by which requires adaptation of HRT firmware, or indirectly by simulating mouse and keyboard messages. Its utilization in real experiments speeded up the operator’s communication with the system cca. Two times in comparison with the traditional control by the mouse and the keyboard.
基金the National Natural Science Foundation of China(Nos.62275164,61905145,and 62275168)National Key Research and Development Program of China(No.2022YFA1200116)+1 种基金Guangdong Natural Science Foundation and Province Project(No.2021A1515011916)Shenzhen Science and Technology Planning Project(No.ZDSYS20210623092006020).
文摘Optical tweezers that rely on laser irradiation to capture and manipulate nanoparticles have provided powerful tools for biological and biochemistry studies.However,the existence of optical diffraction-limit and the thermal damage caused by high laser power hinder the wider application of optical tweezers in the biological field.For the past decade,the emergence of optothermal tweezers has solved the above problems to a certain extent,while the auxiliary agents used in optothermal tweezers still limit their biocompatibility.Here,we report a kind of nanotweezers based on the sign transformation of the thermophoresis coefficient of colloidal particles in low-temperature environment.Using a self-made microfluidic refrigerator to reduce the ambient temperature to around 0℃in the microfluidic cell,we can control a single nanoparticle at lower laser power without adding additional agent solute in the solution.This novel optical tweezering scheme has provided a new path for the manipulation of inorganic nanoparticles as well as biological particles.
基金supported by the National Natural Science Foundation of China(Grant Nos.62075193,11304282,and 61601405)Joint Fund of Ministry ofEducation,China(Grant No.6141A02011604)+2 种基金Major Scientific Research Project of Zhejiang Lab,China(Grant No.2019MB0AD01)National Program for Special Support of Top-Notch Young Professionals,China(Grant No.W02070390)Fundamental Research Funds for the Central Universities,China(Grant Nos.2016XZZX004-01 and 2018XZZX001-08).
文摘Sensing sensitivity is the key performance of optical tweezers.By adjusting the frequency and magnitude of an applied Coulomb force as an input of optical tweezers,we directly measured the sensitivity and signal-to-noise ratio(SNR)of a system and indirectly calculated the actual noise magnitude.Combined with an output filter,the relationship between the SNR and bandwidths was studied.We established the simulation model of a system using Simulink and simulated the relationship between the SNR and magnitude of the input forces and filter bandwidths.In addition,we built an experimental system to determine the relationship between the SNR and the magnitude of the input forces and filter bandwidths.The actual minimum detectable force was measured as 1.8275×10^(-17)N at a 1Hz bandwidth.The experimental results were correlated with the simulation and theoretical results,confirming the effectiveness of the proposed method and demonstrating the high sensitivity of vacuum optical tweezers as mechanical sensors.We proposed a novel method of calibration and measurement of system sensing parameters by applying an actual force that was more direct and precise than the theoretical calculation method that requires accurate fitting parameters,such as the particle radius and density.This method can be employed to analyze the system noise and phase characteristics to confirm and improve the real performance of the system.
文摘In recent years,levitated particles of optical traps in vacuum have shown the enormous potential for precision sensor development and new physics exploration.However,the accuracy of the sensor is still hampered by the uncertainty of the calibration factor relating the detected signal to the absolute displacement of the trapped particle.In this paper,we suggest and experimentally demonstrate a novel calibration method for optical tweezers based on free-falling particles in vacuum,where the gravitational acceleration is introduced as an absolute reference.Our work provides a calibration protocol with a great certainty and traceability,which is significant in improving the accuracy of precision sensing based on levitated optomechanical systems.
基金Project supported by the National Key Research and Development Program of China (Grant No.2020YFA0211400)the State Key Program of the National Natural Science Foundation of China (Grant No.11834008)+3 种基金the National Natural Science Foundation of China (Grant Nos.12174192 and 12204119)the Fund from the State Key Laboratory of Acoustics,Chinese Academy of Sciences (Grant No.SKLA202210)the Fund from the Key Laboratory of Underwater Acoustic Environment,Chinese Academy of Sciences (Grant No.SSHJ-KFKT-1701)the Science and Technology Foundation of Guizhou Province,China (Grant No.ZK[2023]249)。
文摘Acoustic radiation force(ARF), as an important particle manipulation method, has been extensively studied in recent years. With the introduction of the concept of “acoustic tweezers”, negative acoustic radiation has become a research hotspot. In this paper, a scheme of realizing negative ARF based on the multiple-layered spherical structure design is proposed. The specific structure and design idea are presented. Detailed theoretical calculation analysis is carried out.Numerical simulations have been performed to verify the correctness of this prediction. The conjecture that the suppression of backscattering can achieve negative ARF is verified concretely, which greatly expands the application prospect and design ideas of the ARF. This work has laid a theoretical foundation for realizing precise control of the structure.
基金the National Science Foundation(No.NSF-CMMI-1761743)the National Aeronautics and Space Administration Early Career Faculty Award(No.80NSSC17K0520)+2 种基金the National Institute of General Medical Sciences of the National Institutes of Health(No.DP2GM128446)L.H.L.acknowledges financial support from the National Natural Science Foundation of China(No.62075111)the State Key Laboratory of Precision Measurement Technology and Instruments.The authors are grateful to Prof.Brian A.Korgel and Dr.Taizhi Jiang for providing Si particles.They also thank Yaoran Liu,Jingang Li,Kan Yao and Zhihan Chen for useful discussions。
文摘Optical manipulation of micro/nanoscale objects is of importance in life sciences,colloidal science,and nanotechnology.Optothermal tweezers exhibit superior manipulation capability at low optical intensity.However,our implicit understanding of the working mechanism has limited the further applications and innovations of optothermal tweezers.Herein,we present an atomistic view of opto-thermo-electro-mechanic coupling in optothermal tweezers,which enables us to rationally design the tweezers for optimum performance in targeted applications.Specifically,we have revealed that the non-uniform temperature distribution induces water polarization and charge separation,which creates the thermoelectric field dominating the optothermal trapping.We further design experiments to systematically verify our atomistic simulations.Guided by our new model,we develop new types of optothermal tweezers of high performance using low-concentrated electrolytes.Moreover,we demonstrate the use of new tweezers in opto-thermophoretic separation of colloidal particles of the same size based on the difference in their surface charge,which has been challenging for conventional optical tweezers.With the atomistic understanding that enables the performance optimization and function expansion,optothermal tweezers will further their impacts.
