Label-free single-cell isolation enabled by microfluidic impact printing and real-time cellular recognition

Wang, Yiming; Wang, Xiaojie; Pan, Tingrui; Li, Baoqing; Chu, Jiaru

doi:10.1039/d1lc00326g

Cited by 19 publications

(22 citation statements)

References 53 publications

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“…If the target single cell is identified, a piezoelectric actuator is triggered to strike a flexible membrane on the printer chamber to generate a droplet within the target single cell to the defined location. In the demonstration, this single-cell printer printed the HeLa cell with an efficiency of 90.3% at a throughput of 2 Hz, and the cell viability reached 96.6% after printing [41].…”

Section: Label-free Computer Vision-based Single-cell Printingmentioning

confidence: 98%

“…Furthermore, single-cell bioprinting with the inkjet printing method could be achieved only by diluting the cell suspension to reduce the cell density [80,82,83]. Therefore, to increase the reliability and efficiency of single-cell printing, various new techniques have been developed to improve the dispensing efficiency of inkjet-like single-cell printing [40][41][42][44][45][46]68,[84][85][86][87][88][89][90][91][92][93][94][95][96][97][98][99].…”

Section: Noncontact Printingmentioning

confidence: 99%

“…Besides the acoustic field-based inkjet-like single-cell printing, the other popular and useful strategy used to increase the reliability and efficiency of single-cell printing is integrating real-time monitoring techniques with a microfluidic dispenser or printer [32,[40][41][42]97,98,106]. These monitoring techniques can detect and monitor the cells to be printed and trigger the integrated microfluidic dispenser or printer to generate and print droplets containing single cells.…”

Section: Label-free Computer Vision-based Single-cell Printingmentioning

confidence: 99%

“…Recently, Chu and co-workers developed another computer vision-based highly efficient single-cell printer by integrating real-time cell recognition and microfluidic impact printing (Figure 3b) [41]. This single-cell printer consists of a printing module, a signal control module, and an imaging processing module (Figure 3b).…”

Section: Label-free Computer Vision-based Single-cell Printingmentioning

confidence: 99%

“…Second, the printed single cells or colonies can be easily recovered with addressability for subsequent analysis. Third, it is convenient to integrate the highly efficient single-cell printing with other techniques, such as imaging system [40,41], electric field [42][43][44], and acoustic field [45,46], and the single-cell encapsulation efficiency can reach more than 90%. By comparison, the theoretical limit of the single-cell capture efficiency of the widely used droplet-based microfluidic approach is only 37% according to Poisson's distribution.…”

Section: Introductionmentioning

confidence: 99%

See 4 more Smart Citations

Advances in Single-Cell Printing

Zhou¹,

Wu²,

Wen³

et al. 2022

Micromachines

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Single-cell analysis is becoming an indispensable tool in modern biological and medical research. Single-cell isolation is the key step for single-cell analysis. Single-cell printing shows several distinct advantages among the single-cell isolation techniques, such as precise deposition, high encapsulation efficiency, and easy recovery. Therefore, recent developments in single-cell printing have attracted extensive attention. We review herein the recently developed bioprinting strategies with single-cell resolution, with a special focus on inkjet-like single-cell printing. First, we discuss the common cell printing strategies and introduce several typical and advanced printing strategies. Then, we introduce several typical applications based on single-cell printing, from single-cell array screening and mass spectrometry-based single-cell analysis to three-dimensional tissue formation. In the last part, we discuss the pros and cons of the single-cell strategies and provide a brief outlook for single-cell printing.

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Section: Label-free Computer Vision-based Single-cell Printingmentioning

confidence: 98%

Section: Noncontact Printingmentioning

confidence: 99%

Section: Label-free Computer Vision-based Single-cell Printingmentioning

confidence: 99%

Section: Label-free Computer Vision-based Single-cell Printingmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Advances in Single-Cell Printing

Zhou¹,

Wu²,

Wen³

et al. 2022

Micromachines

View full text Add to dashboard Cite

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Harnessing the Manipulation of Single Cells to Construct Biological Structures: Tools and Applications

Liu,

Chen,

Tong

et al. 2024

Adv Funct Materials

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Artificial biological structures hold the promise for modeling cellular assembly in vitro and have advanced considerable studies in cell biology, disease modeling, drug testing, and regenerative medicine. Biological functions are derived from micro‐ and macroscale interactions of various cell types, and a structural property matching the tissue in vivo is required to enable precision biological function. Despite various types of tissues and organs are successfully constructed by conventional biofabrication technologies, they mostly only show a small fraction of structural features found in real tissues. Tools for single‐cell manipulation provide the approach to fabricate artificial tissues cell‐by‐cell, and have enabled the construction of biological structures with single‐cell and heterogeneous features, recapitulating the complexity in vivo. This review presents a comprehensive overview of the construction of biological structures through manipulating single cells, covering single‐cell technologies with operation principles and main advances, biological structures associated with informative explanations of single‐cell manipulation during construction, and representative applications mainly focusing on analysis and modeling. Current challenges and future perspectives in this field are also discussed.

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A Microfluidic Chip on a Robotic Manipulator for Loading and Reloading of Oocytes

Liang,

Amaya,

Sugiura

et al. 2024

Advanced Intelligent Systems

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Loading individual oocytes is a critical step for injecting RNA, expressing heterologous proteins, performing electrophysiological measurements, and so on. However, existing methods remain a challenge for automatically loading multiple single oocytes into different locations. Herein, a novel microfluidic chip on a robotic manipulator (chip‐on‐robot) with feedback control for flexible manipulation of multiple oocytes within a large spatial range is proposed. The manipulator automatically controls the microfluidic chip to reach different locations based on imaging feedback. The microfluidic chip then utilizes the hydrodynamic focusing effect of the main channel to separate oocytes for individual loading or reloading under capacitive sensor feedback. The separation distance reaches approximately 16 times the oocyte diameter. Moreover, capacitive signal feedback on the number of oocytes for flow direction control ensures the separation of all oocytes. For close‐loop control of the loading/reloading process, image‐based oocyte detection is combined using deep learning to calculate the target position of the oocyte. Finally, an automatic sequence is achieved to load multiple single oocytes into a well chip by using the chip‐on‐robot. As a demonstration, the oocytes are reloaded into a specified location based on the conditions. The proposed chip‐on‐robot with feedback control has significant advantages in the micromanipulation of oocytes.

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Label-free single-cell isolation enabled by microfluidic impact printing and real-time cellular recognition

Abstract: Analysis of cellular components at single-cell level is important to reveal cellular heterogeneity. However, current technologies to isolate individual cells are either label-based or with low performance. Here, we present...

Cited by 19 publications

References 53 publications

Advances in Single-Cell Printing

Advances in Single-Cell Printing

Harnessing the Manipulation of Single Cells to Construct Biological Structures: Tools and Applications

A Microfluidic Chip on a Robotic Manipulator for Loading and Reloading of Oocytes

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