scholarly journals Teflon-on-Glass Molding Enables High-Throughput Fabrication of Hydrophilic-in-Hydrophobic Microwells for Bead-Based Digital Bioassays

Materials ◽  
2018 ◽  
Vol 11 (11) ◽  
pp. 2154
Author(s):  
Lisa Tripodi ◽  
Karen Ven ◽  
Dries Kil ◽  
Iene Rutten ◽  
Robert Puers ◽  
...  

In recent years, Teflon-on-glass microwells have been successfully implemented in bead-based digital bioassays for the sensitive detection of single target molecules. Their hydrophilic-in-hydrophobic (HIH) nature enables the isolation and analysis of individual beads, carrying the target molecules, which can be further manipulated accurately through optical tweezer (OT) setups. However, these Teflon HIH-microwell platforms are conventionally fabricated through a complex, time-consuming and labor-intensive dry lift-off procedure which involves a series of major steps, limiting the up-scaling potential of these platforms. Alternative Teflon-based microwell fabrication methods have been extensively explored in literature but they preclude the generation of hydrophobic wells with hydrophilic bottom, thereby hampering the bioassay performance. Here, we present a new Teflon-on-glass molding method for the high throughput fabrication of hydrophilic-in-hydrophobic (HIH) microwell arrays, able to empower bead-based digital bioassays. Microwells 2.95 μm in depth and 3.86 μm in diameter were obtained to host individual beads. In these microwell arrays, sealing of reagents was demonstrated with an efficiency of 100% and seeding of superparamagnetic beads was achieved with an efficiency of 99.6%. The proposed method requires half as many steps when compared to the traditional dry lift-off process, is freely scalable and has the potential to be implemented in different bead-based bioassay applications.

2018 ◽  
Vol 26 (26) ◽  
pp. 34665 ◽  
Author(s):  
Zongbao Li ◽  
Jianxin Yang ◽  
Shaojing Liu ◽  
Xiaofang Jiang ◽  
Haiyan Wang ◽  
...  

2006 ◽  
Vol 46 (4) ◽  
pp. 1549-1562 ◽  
Author(s):  
Justin Klekota ◽  
Erik Brauner ◽  
Frederick P. Roth ◽  
Stuart L. Schreiber

2020 ◽  
Vol 6 (43) ◽  
pp. eabb7438
Author(s):  
Jihwan Lee ◽  
Zhuohe Liu ◽  
Peter H. Suzuki ◽  
John F. Ahrens ◽  
Shujuan Lai ◽  
...  

Unraveling the genetic and epigenetic determinants of phenotypes is critical for understanding and re-engineering biology and would benefit from improved methods to separate cells based on phenotypes. Here, we report SPOTlight, a versatile high-throughput technique to isolate individual yeast or human cells with unique spatiotemporal profiles from heterogeneous populations. SPOTlight relies on imaging visual phenotypes by microscopy, precise optical tagging of single target cells, and retrieval of tagged cells by fluorescence-activated cell sorting. To illustrate SPOTlight’s ability to screen cells based on temporal properties, we chose to develop a photostable yellow fluorescent protein for extended imaging experiments. We screened 3 million cells expressing mutagenesis libraries and identified a bright new variant, mGold, that is the most photostable yellow fluorescent protein reported to date. We anticipate that the versatility of SPOTlight will facilitate its deployment to decipher the rules of life, understand diseases, and engineer new molecules and cells.


2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Dae-Myeong Geum ◽  
SangHyeon Kim ◽  
Seong Kwang Kim ◽  
SooSeok Kang ◽  
JiHoon Kyhm ◽  
...  

AbstractIn this study, multicolor photodetectors (PDs) fabricated by using bulk p-i-n-based visible GaAs and near-infrared InGaAs structures were monolithically integrated through a high-throughput epitaxial lift-off (ELO) process. To perform multicolor detection in integrated structures, GaAs PDs were transferred onto InGaAs PDs by using a Y2O3 bonding layer to simultaneously detect visible and near-infrared photons and minimize the optical loss. As a result, it was found that the GaAs top PD and InGaAs bottom PD were vertically aligned without tilting in x-ray diffraction (XRD) measurement. A negligible change in the dark currents for each PD was observed in comparison with reference PDs through electrical characterization. Furthermore, through optical measurements and simulation, photoresponses were clearly revealed in the visible and near-infrared band for the material’s absorption region, respectively. Finally, we demonstrated the simultaneous multicolor detection of the visible and near-infrared region,which implies individual access to each PD without mutual interference. These results are a significant improvement for the fabrication of multicolor PDs that enables the formation of bulk-based multicolor PDs on a single substrate with a high pixel density and nearly perfect vertical alignment for high-resolution multicolor imaging.


2012 ◽  
Vol 27 (26) ◽  
pp. 3905-3921 ◽  
Author(s):  
Kaushal K. Garg ◽  
Suhas P. Wani ◽  
Jennie Barron ◽  
Louise Karlberg ◽  
Johan Rockstrom

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