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Freiburg, Germany

Leibacher I.,ETH Zurich | Schoendube J.,Albert Ludwigs University of Freiburg | Schoendube J.,Cytena GmbH | Dual J.,ETH Zurich | And 3 more authors.
Biomicrofluidics | Year: 2015

Recent years have witnessed a strong trend towards analysis of single-cells. To access and handle single-cells, many new tools are needed and have partly been developed. Here, we present an improved version of a single-cell printer which is able to deliver individual single cells and beads encapsulated in free-flying picoliter droplets at a single-bead efficiency of 96% and with a throughput of more than 10 beads per minute. By integration of acoustophoretic focusing, the cells could be focused in x and y direction. This way, the cells were lined-up in front of a 40??m nozzle, where they were analyzed individually by an optical system prior to printing. In agreement with acoustic simulations, the focusing of 10??m beads and Raji cells has been achieved with an efficiency of 99% (beads) and 86% (Raji cells) to a 40??m wide center region in the 1?mm wide microfluidic channel. This enabled improved optical analysis and reduced bead losses. The loss of beads that ended up in the waste (because printing them as single beads arrangements could not be ensured) was reduced from 52% ±?6% to 28% ± 1%. The piezoelectric transducer employed for cell focusing could be positioned on an outer part of the device, which proves the acoustophoretic focusing to be versatile and adaptable. © 2015 AIP Publishing LLC.

Gross A.,Albert Ludwigs University of Freiburg | Gross A.,Cytena GmbH | Schoendube J.,Albert Ludwigs University of Freiburg | Schoendube J.,Cytena GmbH | And 5 more authors.
International Journal of Molecular Sciences | Year: 2015

The handling of single cells is of great importance in applications such as cell line development or single-cell analysis, e.g., for cancer research or for emerging diagnostic methods. This review provides an overview of technologies that are currently used or in development to isolate single cells for subsequent single-cell analysis. Data from a dedicated online market survey conducted to identify the most relevant technologies, presented here for the first time, shows that FACS (fluorescence activated cell sorting) respectively Flow cytometry (33% usage), laser microdissection (17%), manual cell picking (17%), random seeding/dilution (15%), and microfluidics/lab-on-a-chip devices (12%) are currently the most frequently used technologies. These most prominent technologies are described in detail and key performance factors are discussed. The survey data indicates a further increasing interest in single-cell isolation tools for the coming years. Additionally, a worldwide patent search was performed to screen for emerging technologies that might become relevant in the future. In total 179 patents were found, out of which 25 were evaluated by screening the title and abstract to be relevant to the field. © 2015 by the authors; licensee MDPI, Basel, Switzerland.

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