Mechanical Characterization of Vesicles and Cells: A Review

Adnan Morshed Buddini Iroshika Karawdeniya Y.M. Nuwan D.Y. Bandara Min Jun Kim Prashanta Dutta

Vesicles perform many essential functions in all living organisms. They respond like a transducer to mechanical stress in converting the applied force into mechanical and biological responses. At the same time, both biochemical and biophysical signals influence the vesicular response in bearing mechanical loads. In recent years, liposomes, artificial lipid vesicles, have gained substantial attention from the pharmaceutical industry as a prospective drug carrier which can also serve as an artificial cell‐mimetic system. The ability of these vesicles to enter through pores of even smaller size makes them ideal candidates for therapeutic agents to reach the infected sites effectively. Engineering of vesicles with desired mechanical properties that can encapsulate drugs and release as required is the prime challenge in this field. This requirement has led to the modifications of the composition of the bilayer membrane by adding cholesterol, sphingomyelin, etc. In this paper, we review the manufacturing and characterization techniques of various artificial/synthetic vesicles. We particularly focus on the electric field‐driven characterization techniques to determine different properties of vesicle and its membranes such as bending rigidity, viscosity, capacitance, conductance, etc., which are indicators of their content and mobility. Similarities and differences between artificial vesicles, natural vesicles, and cells are highlighted throughout the manuscript since most of these artificial vesicles are intended for cell mimetic functions.

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On‐chip technology for single‐cell arraying, electrorotation‐based analysis and selective release

Kevin Keim, Mohamed Z. Rashed, Samuel C. Kilchenmann, Aurélien Delattre, António F. Gonçalves, Paul Éry, Carlotta Guiducci

This paper reports a method for label‐free single‐cell biophysical analysis of multiple cells trapped in suspension by electrokinetic forces. Tri‐dimensional pillar electrodes arranged along the width of a microfluidic chamber define actuators for single cell trapping and selective release by electrokinetic force. Moreover, a rotation can be induced on the cell in combination with a negative DEP force to retain the cell against the flow. The measurement of the rotation speed of the cell as a function of the electric field frequency define an electrorotation spectrum that allows to study the dielectric properties of the cell. The system presented here shows for the first time the simultaneous electrorotation analysis of multiple single cells in separate micro cages that can be selectively addressed to trap and/or release the cells. Chips with 39 micro‐actuators of different interelectrode distance were fabricated to study cells with different sizes. The extracted dielectric properties of Henrietta Lacks, human embryonic kidney 293, and human immortalized T lymphocytes cells were found in agreements with previous findings. Moreover, the membrane capacitance of M17 neuroblastoma cells was investigated and found to fall in in the range of 7.49 ± 0.39 mF/m2.

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Laser interference‐based technique for dynamic measurement of single cell deformation manipulated by optical tweezers

Jiaqi Liu, Fan Zhang, Lianqing Zhu, Xinghua Qu, Daping Chu

A laser interference‐based method was proposed to measure the deformation response of cell manipulated by optical tweezers. This method was implemented experimentally by integrating a laser illuminating system and optical tweezers with an inverted microscope. Interference fringes generated by the transmitted and reflected lights were recorded by a complementary metal oxide semiconductor camera. From the acquired images, cell height was calculated and cell morphology was constructed. To further validate this method, the morphological analyses of HeLa cells were performed in static state and during detachment process. Subsequently, the dynamic deformation responses of red blood cells were measured during manipulation with optical tweezers. Collectively, this laser interference‐based method precludes the requirement of complex optical alignment, allows easy integration with optical tweezers, and enables dynamic measurement of cell deformation response by using a conventional inverted microscope.

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Hybrid microfluidics combined with active and passive approaches for continuous cell separation

Sheng Yan, Jun Zhang, Dan Yuan, Weihua Li

Microfluidics, which is classified as either active or passive, is capable of separating cells of interest from a complex and heterogeneous sample. Active methods utilise external fields such as electric, magnetic, acoustic, and optical to drive cells for separation, while passive methods utilise channel structures, intrinsic hydrodynamic forces, and steric hindrances to manipulate cells. However, when processing complex biological samples such as whole blood with rare cells, separation with a single module microfluidic device is difficult. Hybrid microfluidics is an emerging technique which utilises active and passive methods whilst fulfilling higher requirements for stable performance, versatility, and convenience, including (i) the ability to process multi-target cells, (ii) enhanced ability for multiplexed separation, (iii) higher sensitivity, and (iv) tunability for a wider operational range. This review introduces the fundamental physics and typical formats for subclasses of hybrid microfluidic devices based on their different physical fields; presents current examples of cell sorting to highlight the advantage and usefulness of hybrid microfluidics on biomedicine, and then discusses the challenges and perspective of future development and the promising direction of research in this field.

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Joule heating monitoring in a microfluidic channel by observing the Brownian motion of an optically trapped microsphere

Toon Brans, Filip Strubbe, Caspar Schreuer, Stijn Vandewiele, Kristiaan Neyts and Filip Beunis

Electric fields offer a variety of functionalities to Lab-on-a-Chip devices. The use of these fields often results in significant Joule heating, affecting the overall performance of the system. Precise knowledge of the temperature profile inside a microfluidic device is necessary to evaluate the implications of heat dissipation. This article demonstrates how an optically trapped microsphere can be used as a temperature probe to monitor Joule heating in these devices. The Brownian motion of the bead at room temperature is compared with the motion when power is dissipated in the system. This gives an estimate of the temperature increase at a specific location in a microfluidic channel. We demonstrate this method with solutions of different ionic strengths, and establish a precision of 0.9 K and an accuracy of 15%. Furthermore it is demonstrated that transient heating processes can be monitored with this technique, albeit with a limited time resolution.

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Optical manipulation of charged microparticles in polar fluids

Giuseppe Pesce, Vincenzo Lisbino, Giulia Rusciano, Antonio Sasso

In this study, we report a systematic study of the response of a charged microparticle confined in an optical trap and driven by electric fields. The particle is embedded in a polar fluid, hence, the role of ions and counterions forming a double layer around the electrodes and the particle surface itself has been taken into account. We analyze two different cases: (i) electrodes energized by a step-wise voltage (DC mode) and (ii) electrodes driven by a sinusoidal voltage (AC mode). The experimental outcomes are analyzed in terms of a model that combines the electric response of the electrolytic cell and the motion of the trapped particle. In particular, for the DC mode we analyze the transient particle motion and correlate it with the electric current flowing in the cell. For the AC mode, the stochastic and deterministic motion of the trapped particle is analyzed either in the frequency domain (power spectral density, PSD) or in the time domain (autocorrelation function). Moreover, we will show how these different approaches (DC and AC modes) allow us, assuming predictable the applied electric field (here generated by plane parallel electrodes), to provide accurate estimation (3%) of the net charge carried by the microparticle. Vice versa, we also demonstrate how, once predetermined the charge, the trapped particle acts as a sensitive probe to reveal locally electric fields generated by arbitrary electrode geometries (in this work, wire-tip geometry).
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Mapping AC Electroosmotic Flow at the Dielectrophoresis Crossover Frequency of a Colloidal Probe

Jingyu Wang, Ming-Tzo Wei, Joel A. Cohen, H. Daniel Ou-Yang
AC electroosmotic (ACEO) flow above the gap between coplanar electrodes is mapped by the measurement of Stokes forces on an optically-trapped polystyrene colloidal particle. E2-dependent forces on the probe particle are selected by amplitude modulation (AM) of the ACEO electric field (E) and lock-in detection at twice the AM frequency. E2-dependent dielectrophoresis (DEP) of the probe is eliminated by driving the ACEO at the probe's DEP crossover frequency. The location-independent DEP crossover frequency is determined, in a separate experiment, as the limiting frequency of zero horizontal force as the probe is moved toward the midpoint between the electrodes. The ACEO velocity field, uncoupled from probe DEP effects, was mapped in the region 1–9 μm above a 28 μm gap between the electrodes. By use of variously-sized probes, each at its DEP crossover frequency, the frequency dependence of the ACEO flow was determined at a point 3 μm above the electrode gap and 4 μm from an electrode tip. At this location the ACEO flow was maximal at ∼117 kHz for a low-salt solution. This optical trapping method, by eliminating DEP forces on the probe, provides unambiguous mapping of the ACEO velocity field.
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Orthogonal optical force separation simulation of particle and molecular species mixtures under direct current electroosmotic driven flow for applications in biological sample preparation

Sarah J. R. Staton, Alex Terray, Greg E. Collins, Sean J. Hart

Presented here are the results from numerical simulations applying optical forces orthogonally to electroosmotically induced flow containing both molecular species and particles. Simulations were conducted using COMSOL v4.2a Multiphysics® software including the particle tracking module. The study addresses the application of optical forces to selectively remove particulates from a mixed sample stream that also includes molecular species in a pinched flow microfluidic device. This study explores the optimization of microfluidic cell geometry, magnitude of the applied direct current electric field, electroosmotic flow rate, diffusion, and magnitude of the applied optical forces. The optimized equilibrium of these various contributing factors aids in the development of experimental conditions and geometry for future experimentation as well as directing experimental expectations, such as diffusional losses, separation resolution, and percent yield. The result of this work generated an optimized geometry with flow conditions leading to negligible diffusional losses of the molecular species while also being able to produce particle removal at near 100% levels. An analytical device, such as the one described herein with the capability to separate particulate and molecular species in a continuous, high-throughput fashion would be valuable by minimizing sample preparation and integrating gross sample collection seamlessly into traditional analytical detection methods.

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Dielectrophoresis force spectroscopy for colloidal clusters

Hyunjoo Park, Ming-Tzo Wei, H. Daniel Ou-Yang

Optical trapping-based force spectroscopy was used to measure the frequency dependent dielectrophoresis (DEP) forces and DEP crossover frequencies of colloidal PMMA spheres and clusters. A single sphere or cluster, held by an optical tweezer, was positioned near the center of a pair of gold-film electrodes where ACEO flow was negligible. Use of amplitude modulation and phase-sensitive lock-in detection for accurate measurement of the DEP force yielded new insight into dielectric relaxation mechanisms near the crossover frequencies. On one hand, the size dependence of the DEP force near the crossover frequencies indicates that the dominant polarization mechanism is a volume effect. On the other, the power-law dependence of the crossover frequency on the particle radius with an exponent -2 indicates the dielectric relaxation is more likely due to ionic diffusion across the particle surface, suggesting the dominant polarization mechanism may be a surface polarization effect. Better theories are needed to explain the experiment. Nevertheless, the strong size dependence of the crossover frequencies suggests the use of DEP for size sorting of micron-sized particles.

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Light at work: The use of optical forces for particle manipulation, sorting, and analysis

Alexandr Jonás, Pavel Zemánek

We review the combinations of optical micro-manipulation with other techniques and their classical and emerging applications to non-contact optical separation and sorting of micro- and nanoparticle suspensions, compositional and structural analysis of specimens, and quantification of force interactions at the microscopic scale. The review aims at inspiring researchers, especially those working outside the optical micro-manipulation field, to find new and interesting applications of these methods.

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