Label-free purification and characterization of optogenetically engineered cells using optically-induced dielectrophoresis

Yang, Jia; Gu, Yanyu; Zhang, Chuang; Zhang, Yuzhao; Hao, Liqiang; Zhao, Ying; Liu, Lianqing; Wang, Wenxue

doi:10.1039/d2lc00512c

Cited by 8 publications

(3 citation statements)

References 61 publications

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“…[ 19 ] The OET system has been utilized to extract and purify circulating tumor cells (CTCs) from blood samples, [ 20 ] separate live gastric cancer cells from patients' complete ascites, [ 21 ] and purify optogenetically engineered cells. [ 22 ] However, recent OET‐based devices have predominantly focused on direct cell sorting, with few instances of forming high‐throughput single‐cell arrays assisted by a fixed structure (micro‐well array). Our newly designed OET chip is an integrated, multifunctional platform capable of performing single‐cell sorting and retrieval.…”

Section: Introductionmentioning

confidence: 99%

Optoelectronic Tweezers Micro‐Well System for Highly Efficient Single‐Cell Trapping, Dynamic Sorting, and Retrieval

Gan,

Zhang,

Chen

et al. 2024

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Single‐cell arrays have emerged as a versatile method for executing single‐cell manipulations across an array of biological applications. In this paper, an innovative microfluidic platform is unveiled that utilizes optoelectronic tweezers (OETs) to array and sort individual cells at a flow rate of 20 µL min−1. This platform is also adept at executing dielectrophoresis (DEP)‐based, light‐guided single‐cell retrievals from designated micro‐wells. This presents a compelling non‐contact method for the rapid and straightforward sorting of cells that are hard to distinguish. Within this system, cells are individually confined to micro‐wells, achieving an impressive high single‐cell capture rate exceeding 91.9%. The roles of illuminating patterns, flow velocities, and applied electrical voltages are delved into in enhancing the single‐cell capture rate. By integrating the OET system with the micro‐well arrays, the device showcases adaptability and a plethora of functions. It can concurrently trap and segregate specific cells, guided by their dielectric signatures. Experimental results, derived from a mixed sample of HepG2 and L‐O2 cells, reveal a sorting accuracy for L‐O2 cells surpassing 91%. Fluorescence markers allow for the identification of sequestered, fluorescence‐tagged HepG2 cells, which can subsequently be selectively released within the chip. This platform's rapidity in capturing and releasing individual cells augments its potential for future biological research and applications.

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Section: Introductionmentioning

confidence: 99%

Optoelectronic Tweezers Micro‐Well System for Highly Efficient Single‐Cell Trapping, Dynamic Sorting, and Retrieval

Gan,

Zhang,

Chen

et al. 2024

Small

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“…To manipulate cells in a liquid environment, various micromanipulation techniques, including optical tweezers [ 37 – 39 ], optoelectronic tweezers [ 40 , 41 ], acoustic tweezers [ 42 – 44 ], dielectrophoresis (DEP) [ 45 , 46 ], and magnetic tweezers [ 47 – 49 ], have been developed. For example, optical tweezers generate a local light field gradient on the surface or inside the cell by focusing the laser beam.…”

Section: Introductionmentioning

confidence: 99%

Integrated Cross-Scale Manipulation and Modulable Encapsulation of Cell-Laden Hydrogel for Constructing Tissue-Mimicking Microstructures

Zhao,

Dong,

et al. 2024

Research

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Engineered microstructures that mimic in vivo tissues have demonstrated great potential for applications in regenerative medicine, drug screening, and cell behavior exploration. However, current methods for engineering microstructures that mimic the multi-extracellular matrix and multicellular features of natural tissues to realize tissue-mimicking microstructures in vitro remain insufficient. Here, we propose a versatile method for constructing tissue-mimicking heterogeneous microstructures by orderly integration of macroscopic hydrogel exchange, microscopic cell manipulation, and encapsulation modulation. First, various cell-laden hydrogel droplets are manipulated at the millimeter scale using electrowetting on dielectric to achieve efficient hydrogel exchange. Second, the cells are manipulated at the micrometer scale using dielectrophoresis to adjust their density and arrangement within the hydrogel droplets. Third, the photopolymerization of these hydrogel droplets is triggered in designated regions by dynamically modulating the shape and position of the excitation ultraviolet beam. Thus, heterogeneous microstructures with different extracellular matrix geometries and components were constructed, including specific cell densities and patterns. The resulting heterogeneous microstructure supported long-term culture of hepatocytes and fibroblasts with high cell viability (over 90%). Moreover, the density and distribution of the 2 cell types had significant effects on the cell proliferation and urea secretion. We propose that our method can lead to the construction of additional biomimetic heterogeneous microstructures with unprecedented potential for use in future tissue engineering applications.

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“…In addition to its high-throughput characteristics [ 22 ] (being able to form up to 50,000 virtual electrodes simultaneously [ 18 ]), OETs merely require 1/500 of optical power to generate traps with stiffness similar to OTs [ 23 ], which demonstrates its high efficiency and noninvasive features for intricate cell manipulation tasks. Furthermore, the mechanism of ODEP has been extensively utilized for modular assembly [ 24 ], cell rotation [ 25 ], actuation and control of microrobot [ 26 , 27 ], cell purification and characterization [ 28 ], etc., which are comprehensively discussed in [ 29 , 30 ]. In recent years, researchers have attempted to utilize programmable schemes [ 19 , 31 – 33 ] based on the ODEP mechanism to facilitate the efficiency and accuracy of micromanipulation.…”

Section: Introductionmentioning

confidence: 99%

POMDP-Based Real-Time Path Planning for Manipulation of Multiple Microparticles via Optoelectronic Tweezers

et al. 2022

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With high throughput and high flexibility, optoelectronic tweezers (OETs) hold huge potential for massively parallel micromanipulation. However, the trajectory of the virtual electrode has been planned in advance in most synchronous manipulations for multiple targets based on an optically induced dielectrophoresis (ODEP) mechanism, which is insufficient to ensure the stability and efficiency in an environment with potential collision risk. In this paper, a synchronously discretized manipulation method based on a centralized and decoupled path planner is proposed for transporting microparticles of different types with an OET platform. An approach based on the Kuhn-Munkres algorithm is utilized to achieve the goal assignment between target microparticles and goal positions. With the assistance of a visual feedback module, a path planning approach based on the POMDP algorithm dynamically determines the motion strategies of the particle movement to avoid potential collisions. The geometrical parameters of the virtual electrodes are optimized for different types of particles with the goal of maximum transport speed. The experiments of micropatterning with different morphologies and transporting multiple microparticles (e.g., polystyrene microspheres and 3T3 cells) to goal positions are performed. These results demonstrate that the proposed manipulation method based on optoelectronic tweezers is effective for multicell transport and promises to be used in biomedical manipulation tasks with high flexibility and efficiency.

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Label-free purification and characterization of optogenetically engineered cells using optically-induced dielectrophoresis

Abstract: Optogenetically engineered cell population obtained by heterogenous gene expression plays a vital role in life science, medicine, and biohybrid robotics, and the purification and characterization are essential to enhance its...

Cited by 8 publications

References 61 publications

Optoelectronic Tweezers Micro‐Well System for Highly Efficient Single‐Cell Trapping, Dynamic Sorting, and Retrieval

Optoelectronic Tweezers Micro‐Well System for Highly Efficient Single‐Cell Trapping, Dynamic Sorting, and Retrieval

Integrated Cross-Scale Manipulation and Modulable Encapsulation of Cell-Laden Hydrogel for Constructing Tissue-Mimicking Microstructures

POMDP-Based Real-Time Path Planning for Manipulation of Multiple Microparticles via Optoelectronic Tweezers

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