Living Cell Microarrays: An Overview of Concepts

Jonczyk, Rebecca; Kurth, Tracy; Lavrentieva, Antonina; Walter, Johanna‐Gabriela; Scheper, Thomas; Stahl, Frank

doi:10.3390/microarrays5020011

Cited by 34 publications

(33 citation statements)

References 188 publications

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“…Additionally, the cell number and density of a given area or volume can be controlled, which allows monitoring of a high spatial and temporal resolution and observation of the dynamic behavior of many cells [93,94]. Parallellisation of experimental conditions and automatization in microfluidic chips has resulted in designs for high-throughput cell screening [95], such as living cell microarrays [96][97][98][99][100][101]. Microfluidic parallelization and single-cell monitoring allows the measurement and averaging of parameters on hundreds of individual cells, compared to measuring parameters of a whole cell population.…”

Section: Microfluidic High-throughput Antifungal Drug Discoverymentioning

confidence: 99%

Micro- and Nanoscale Approaches in Antifungal Drug Discovery

Willaert

2018

Fermentation

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Clinical needs for novel antifungal agents have increased due to the increase of people with a compromised immune system, the appearance of resistant fungi, and infections by unusual yeasts. The search for new molecular targets for antifungals has generated considerable research, especially using modern omics methods (genomics, genome-wide collections of mutants, and proteomics) and bioinformatics approaches. Recently, micro-and nanoscale approaches have been introduced in antifungal drug discovery. Microfluidic platforms have been developed, since they have a number of advantages compared to traditional multiwell-plate screening, such as low reagent consumption, the manipulation of a large number of cells simultaneously and independently, and ease of integrating numerous analytical standard operations and large-scale integration. Automated high-throughput antifungal drug screening is achievable by massive parallel processing. Various microfluidic antimicrobial susceptibility testing (AST) methods have been developed, since they can provide the result in a short time-frame, which is necessary for personalized medicine in the clinic. New nanosensors, based on detecting the nanomotions of cells, have been developed to further decrease the time to test antifungal susceptibility to a few minutes. Finally, nanoparticles (especially, silver nanoparticles) that demonstrated antifungal activity are reviewed.Fermentation 2018, 4, 43 2 of 23 antifungal classes have been reported since 2006 [7]. Current antifungal drugs show some limitations: Amphothericin B (a polyene antibiotic) displays a considerable toxicity and undesirable side effects [8,9], issues with pharmacokinetic properties (such as a short half-life of echinocandins) and activity spectrum, a small number of targets [10,11], and they can interact with other drugs, such as chemotherapy agents and immunosuppressants [12,13]. The last approved antifungal (i.e., anidulafungin) by the European Medicines Agency and the Food and Drug Administration (FDA) dates back to 2006 [14]. There is an urgent need for safer and more effective antifungal drugs. Multiple types of antifungal compounds are in clinical development and these new agents have been recently reviewed [15][16][17].Over the last 20 years, several approaches to antifungal discovery have been explored. The traditional approach seeks, first, to identify active compounds from large compound libraries using a panel of fungal pathogens in standardised assays when possible. In the genetic, genomic, or bioinformatics approach, the objective is, initially, to identify broadly represented targets in fungal pathogens and non-pathogens [18]. In this "target-centric" genomic approach, up-front genetic, bioinformatics, and biochemical target prioritisation is performed and, subsequently, an in vitro-based screening of individual targets is achieved. However, an exceedingly high rate of failure has been observed when applying this approach [19]. Additionally, not all bioactive compounds act through a target-specific mechan...

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Section: Microfluidic High-throughput Antifungal Drug Discoverymentioning

confidence: 99%

Micro- and Nanoscale Approaches in Antifungal Drug Discovery

Willaert

2018

Fermentation

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“…Their methodology uses chemicals to convert the protective groups and at each step of synthesis, nucleotides are fired onto the target spot using nozzles, the same ones found in inkjet printers [29,30]. A major advantage of this technology over the Affymetrix one is that a computer input system controls the synthesis of the oligonucleotides in each microarray.…”

Section: In Situ Synthesized Microarraysmentioning

confidence: 99%

“…A major advantage of this technology over the Affymetrix one is that a computer input system controls the synthesis of the oligonucleotides in each microarray. Although this is very flexible, it is less efficient when making great number of identical microarrays [29,30].…”

Section: In Situ Synthesized Microarraysmentioning

confidence: 99%

DNA Microarray and Bioinformatics Technologies: A mini-review

Missoum¹

2018

Proc. Nat. Res. Soc.

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This bibliographic review gives a concise overview of how DNA microarrays became one of the most important recent technologies. In brief, DNA sequences acting as probes are fixed on tiny slides in arrays to identify the presence of tagged fluorescently nucleic acids from a certain sample. The nucleic acids having complementary sequences with probes will hybridize on the array slides and because they are labeled, they are detected easily using specialized scanner and software. According to the way DNA microarrays are manufactured, there are three main types including spotted arrays, in situ synthesized arrays, and self-assembled arrays. Bioinformatics, which is the use of mathematical and computational methods, plays a crucial role in this technology as it is used for designing specific primers and probes suitable for each sample that is investigated. Moreover, although DNA microarrays are widely used to determine gene expression levels, they are also used in single nucleotide polymorphism (SNP) genotyping. For this application itself, more than one approach can be used including allele discrimination using hybridization, array primer extension assay (APEX), and Illumina infinium assay. The APEX approach was discussed in more detail as it was found that to be useful in screening for inherited genetic diseases such as Stargardt disease. Finally, this review was ended with short discussion of some DNA microarray limitations arising hybridization kinetics as well as difficulties in designing arrays for highly variable genomes.

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“…Cell microarrays provide an attractive solution, as they could increase the throughput significantly [1][2][3]. A cellular microarray consists of a solid support wherein small volumes of different biomolecules and cells can be displayed in defined locations, allowing the multiplexed interrogation of living cells and the analysis of cellular responses [4,5]. Living cell microarrays have been combined with microfluidic bioreactors, which provide multiple advantages for multiplex dynamic analyses and high-throughput screening [4,6].…”

Section: Introductionmentioning

confidence: 99%

Robotic Cell Printing for Constructing Living Yeast Cell Microarrays in Microfluidic Chips

2020

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Living cell microarrays in microfluidic chips allow the non-invasive multiplexed molecular analysis of single cells. Here, we developed a simple and affordable perfusion microfluidic chip containing a living yeast cell array composed of a population of cell variants (green fluorescent protein (GFP)-tagged Saccharomyces cerevisiae clones). We combined mechanical patterning in 102 microwells and robotic piezoelectric cell dispensing in the microwells to construct the cell arrays. Robotic yeast cell dispensing of a yeast collection from a multiwell plate to the microfluidic chip microwells was optimized. The developed microfluidic chip and procedure were validated by observing the growth of GFP-tagged yeast clones that are linked to the cell cycle by time-lapse fluorescence microscopy over a few generations. The developed microfluidic technology has the potential to be easily upscaled to a high-density cell array allowing us to perform dynamic proteomics and localizomics experiments.

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Living Cell Microarrays: An Overview of Concepts

Cited by 34 publications

References 188 publications

Micro- and Nanoscale Approaches in Antifungal Drug Discovery

Micro- and Nanoscale Approaches in Antifungal Drug Discovery

DNA Microarray and Bioinformatics Technologies: A mini-review

Robotic Cell Printing for Constructing Living Yeast Cell Microarrays in Microfluidic Chips

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