Inkjet-like printing of single-cells

Yusof, Azita Laily; Keegan, Helen; Spillane, Cathy; Sheils, Orla; Martín, C; O’Leary, John; Zengerle, Roland; Koltay, Peter

doi:10.1039/c1lc20176j

Cited by 132 publications

(98 citation statements)

References 22 publications

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“…Several approaches have been designed for this purpose, e.g., inkjet cell printing (3)(4)(5)(6), surface engineering (7)(8)(9)(10)(11)(12)(13)(14)(15), and physical constraints (16)(17)(18)(19)(20)(21)(22)(23). However, finding a method that completely satisfies the above requirements remains a challenge.…”

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confidence: 99%

Block-Cell-Printing for live single-cell printing

Zhang¹,

Chou

Xia

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

129

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A unique live-cell printing technique, termed "Block-Cell-Printing" (BloC-Printing), allows for convenient, precise, multiplexed, and high-throughput printing of functional single-cell arrays. Adapted from woodblock printing techniques, the approach employs microfluidic arrays of hook-shaped traps to hold cells at designated positions and directly transfer the anchored cells onto various substrates. BloC-Printing has a minimum turnaround time of 0.5 h, a maximum resolution of 5 μm, close to 100% cell viability, the ability to handle multiple cell types, and efficiently construct protrusion-connected single-cell arrays. The approach enables the large-scale formation of heterotypic cell pairs with controlled morphology and allows for material transport through gap junction intercellular communication. When six types of breast cancer cells are allowed to extend membrane protrusions in the BloC-Printing device for 3 h, multiple biophysical characteristics of cells-including the protrusion percentage, extension rate, and cell length-are easily quantified and found to correlate well with their migration levels. In light of this discovery, BloC-Printing may serve as a rapid and high-throughput cell protrusion characterization tool to measure the invasion and migration capability of cancer cells. Furthermore, primary neurons are also compatible with BloC-Printing.cell array | cell communication | protrusion profiling | neuron patterning C urrent high-throughput screening of cell function and heterogeneity and in vitro cell-cell communication studies requires routine generation of large-scale single-cell arrays with high precision and efficiency, single-cell resolution, multiple cell types, and maintenance of cell viability and function (1, 2). Several approaches have been designed for this purpose, e.g., inkjet cell printing (3-6), surface engineering (7-15), and physical constraints (16-23). However, finding a method that completely satisfies the above requirements remains a challenge. Potentially useful and convenient tools may be available by adapting traditional printing tools to cell printing. In particular, woodblock printing is an efficient and convenient technology that revolutionized the printing world more than 1,800 y ago and was extended to microcontact molecular printing ∼20 y ago (24). However, application of the block-printing concept to cells has never been achieved. The main challenges are (i) inking cells to their molds with precision and maintaining viability, (ii) evenly and gently applying and transferring the cells to a substrate and successfully lifting off the mold without detaching the cells, and (iii) maintaining cell functions after printing.We report here the development and testing of a technology called "Block-Cell-Printing" (BloC-Printing), which involves directly inking cells to a predesigned mold and then transferring the cells to a substrate. We overcome the challenges described above by flow patterning, instead of pressing, the ink objects, as in woodblock or microcontact printing. By ...

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mentioning

confidence: 99%

Block-Cell-Printing for live single-cell printing

Zhang¹,

Chou

Xia

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

129

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“…To date, contact spotting has not been used to array live mammalian cells due to rapid spot evaporation and consequent cell death. Therefore, arraying of live mammalian cells has been limited to inkjet printing, which lacks the ability to handle a large number of different samples 8,9 .Large collections of mammalian cell lines have recently become available, including primary tumor and cancer cell lines 10 , stably transfected expression cell lines, and GFP-fusion libraries 11 . Novel approaches are required to efficiently assemble complex arrays of hundreds to thousands of genetically diverse cells.…”

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confidence: 99%

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confidence: 99%

Live mammalian cell arrays

et al. 2013

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High-content assays have the potential to drastically increase throughput in cell biology and drug discovery, but handling and culturing large libraries of cells such as primary tumor or cancer cell lines requires expensive, dedicated robotic equipment. We have developed a simple, yet powerful method that uses contact spotting to generate highdensity nanowell arrays of live mammalian cells for the culture and interrogation of cell libraries. High-content assays have the potential to drastically increase throughput in cell biology and drug discovery, but handling and culturing large libraries of cells such as primary tumor or cancer cell lines requires expensive, dedicated robotic equipment. We have developed a simple, yet powerful method that uses contact spotting to generate highdensity nanowell arrays of live mammalian cells for the culture and interrogation of cell libraries.Cell-based assays and the tools used to perform them are constantly undergoing improvements toward higher experimental throughput, reduced reagent consumption, and advanced control of the cell microenvironment 1,2 . There are currently two main approaches for conducting highcontent cell culture experiments. In the first, cells are cultured in microtiter plates and robotic equipment is used to perform all necessary fluidic operations 3 . Although effective, this method remains unavailable to many laboratories due to the requirement for expensive robotics. A second approach that can be used for high-throughput cell studies is reverse transfection [4][5][6] . Here, cells are seeded onto an array of DNA, RNA, or small molecules. Large gene expression and silencing studies can be conducted in this manner, but complex cell libraries cannot be investigated since each array is limited to a single cell type.As an alternative to these approaches, contact spotting offers an affordable yet high-throughput platform. This technique is extensively used to generate high-density DNA and protein arrays and has been adapted to a variety of other purposes. For example, Hart et al. created a high-content immunoassay by spotting fixed mammalian cells that were cultured under different conditions 7 . To date, contact spotting has not been used to array live mammalian cells due to rapid spot evaporation and consequent cell death. Therefore, arraying of live mammalian cells has been limited to inkjet printing, which lacks the ability to handle a large number of different samples 8,9 .Large collections of mammalian cell lines have recently become available, including primary tumor and cancer cell lines 10 , stably transfected expression cell lines, and GFP-fusion libraries 11 . Novel approaches are required to efficiently assemble complex arrays of hundreds to thousands of genetically diverse cells. To address this arising need, we developed a simple, fast, and scalable method that uses standard microarray printing tools to generate high-density nanowell arrays. A minimal sample requirement of 500 cells enables the interrogation of cells that are availab...

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“…In the meantime, several works have succeeded in cell ejection using inkjet technology. [10][11][12][13][14][15] Yusof et al 16 have outlined a non-contact method for the controlled separation of single cells confined in a droplet that can be printed inkjet-like onto predefined locations. This method is a suitable platform for printing single cells of various types.…”

Section: Introductionmentioning

confidence: 99%

Accurate dispensing system for single oocytes using air ejection

et al. 2013

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In this study, we propose a new approach to increase the success rate of singleoocyte dispensing and investigate the subsequent viability of the dispensed oocytes. We used a pair of capacitance sensors placed in a microfluidic chip to detect the oocyte, and custom-designed a special buffer zone in the microchannel to decelerate the flow velocity and reduce the hydraulic pressure acting on the oocyte. In the buffer zone, a semicircular bay, formed by equally spaced micropillars, is used to stop the oocyte at the dispensing nozzle hole. Finally, the oocyte is ejected by airflow to the culture array. The novel feature of the developed microfluidic system is that the extraordinary improvement in success rate is accompanied by a lack of change in oocyte survival rate (as assessed by a comparison of survival rates before and after the dispensing procedure). By using this device, we achieved a highly accurate single-oocyte dispensing process with a success rate of 100%. The oocyte survival rate is approximately 70%, regardless of whether or not the oocyte is dispensed. The newly proposed system has the advantages of high operation speed and potential usage for two-dimensional micropatterning. V C 2013 AIP Publishing LLC.

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Inkjet-like printing of single-cells

Cited by 132 publications

References 22 publications

Block-Cell-Printing for live single-cell printing

Block-Cell-Printing for live single-cell printing

Live mammalian cell arrays

Accurate dispensing system for single oocytes using air ejection

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