Cao Y, Zhang W, Liang Y, Feng Z, Jiang C, Chen Z, and Jiang X
Computer Assisted Surgery (Abingdon, England) [Comput Assist Surg (Abingdon)] 2019 Dec; Vol. 24 (1), pp. 1-6. Date of Electronic Publication: 2019 Jan 21.
It is technically demanding and requires rich experience to insert the translaminar facet screw(TFS) via the paramedian mini-incision approach. It seems that it is easy to place the TFS using computer-assisted design and rapid prototyping(RP) techniques. However, the accuracy and safety of these techniques is still unknown. The aim of this study is to assess the accuracy and safety of translaminar facet screw placement in multilevel unilateral transforaminal lumbar interbody fusion using a rapid prototyping drill guide template system. A patient-matched rapid prototyping translaminar facet screw guide was examined in fourteen cadaveric lumbar spine specimens. A three-dimensional (3D) preoperative screw trajectory was constructed using spinal computed tomography scans, from which individualized guides were developed for the placement of translaminar facet screws. Following bone tunnel establishment, the 3D positioning of the entry point and trajectory of the screws was compared to the preoperative plan as found in the Mimics software.Among 60 trajectories eligible for assessment, no cases of clinically significant laminar perforation were found. The mean deviation between the planned and the actual starting points on spinous process was 1.22 mm. The mean tail and submergence angle deviation was found to be 0.68°and 1.46°, respectively. Among all the deviations, none were found to have any statistical significance. These results indicate that translaminar facet screw placement using the guide system is both accurate and safe.
ACS Synthetic Biology [ACS Synth Biol] 2020 Jan 17; Vol. 9 (1), pp. 144-156. Date of Electronic Publication: 2020 Jan 03.
The field of mammalian synthetic biology is expanding quickly, and technologies for engineering large synthetic gene circuits are increasingly accessible. However, for mammalian cell engineering, traditional tissue culture methods are slow and cumbersome, and are not suited for high-throughput characterization measurements. Here we have utilized mammalian cell-free protein synthesis (CFPS) assays using HeLa cell extracts and liquid handling automation as an alternative to tissue culture and flow cytometry-based measurements. Our CFPS assays take a few hours, and we have established optimized protocols for small-volume reactions using automated acoustic liquid handling technology. As a proof-of-concept, we characterized diverse types of genetic regulation in CFPS, including T7 constitutive promoter variants, internal ribosomal entry sites (IRES) constitutive translation-initiation sequence variants, CRISPR/dCas9-mediated transcription repression, and L7Ae-mediated translation repression. Our data shows simple regulatory elements for use in mammalian cells can be quickly prototyped in a CFPS model system.
ACS Applied Materials & Interfaces [ACS Appl Mater Interfaces] 2020 Jan 08; Vol. 12 (1), pp. 1817-1824. Date of Electronic Publication: 2019 Dec 19.
This paper presents a novel method of rapidly customizing microfluidic systems using a consumer-grade inkjet printer and a commercially available superhydrophobic spray. By casting polydimethylsiloxane (PDMS) on liquid templates that are defined by inkjet-printed hydrophilic patterns on superhydrophobically-coated PDMS substrates, microfluidic devices can be directly fabricated. Utilizing the interfacial properties of the superhydrophobic coating and the template liquid, the fabrication of microfluidics could be done with minimum effort and expertise, and unlike previously reported works, no mask and bonding process is necessary. As a proof of concept, we created different microfluidic devices for various applications, like gradient generation and pneumatic control of fluid. Appealing in its simplicity and rapidness, the newly proposed technique could provide an easy-to-use microfluidic platform for front-line researchers with different backgrounds to quickly customize microfluidic devices.
Lehr FX, Hanst M, Vogel M, Kremer J, Göringer HU, Suess B, and Koeppl H
ACS Synthetic Biology [ACS Synth Biol] 2019 Sep 20; Vol. 8 (9), pp. 2163-2173. Date of Electronic Publication: 2019 Aug 27.
RNA-based devices controlling gene expression bear great promise for synthetic biology, as they offer many advantages such as short response times and light metabolic burden compared to protein-circuits. However, little work has been done regarding their integration to multilevel regulated circuits. In this work, we combined a variety of small transcriptional activator RNAs (STARs) and toehold switches to build highly effective AND-gates. To characterize the components and their dynamic range, we used an Escherichia coli (E. coli) cell-free transcription-translation (TX-TL) system dispensed via nanoliter droplets. We analyzed a prototype gate in vitro as well as in silico, employing parametrized ordinary differential equations (ODEs), for which parameters were inferred via parallel tempering, a Markov chain Monte Carlo (MCMC) method. On the basis of this analysis, we created nine additional AND-gates and tested them in vitro. The functionality of the gates was found to be highly dependent on the concentration of the activating RNA for either the STAR or the toehold switch. All gates were successfully implemented in vivo, offering a dynamic range comparable to the level of protein circuits. This study shows the potential of a rapid prototyping approach for RNA circuit design, using cell-free systems in combination with a model prediction.
De Vos J, Dams M, Broeckhoven K, Desmet G, Horstkotte B, and Eeltink S
Analytical Chemistry [Anal Chem] 2020 Jan 13. Date of Electronic Publication: 2020 Jan 13.
A novel multilayer modulator chip offering a robust miniaturized interface for multidimensional liquid chromatography has been developed. The thermoplastic microfluidic device comprises five tailor-made functional layers, and the chip is compatible with commercially available switching-valve technology. The modulator chip allows for robust ultrahigh-pressure operation up to 65 MPa. Peak-dispersion characteristics of system peaks were assessed directly at the valve outlet by monitoring fluorescein injection profiles with laser-induced fluorescence detection. Integration of a microporous monolithic mixing entity in the microchannels significantly narrows the resulting peak profile. Proof-of-concept of the applicability of the microfluidic modulator chip is demonstrated in a heart-cut multidimensional strong-cation-exchange-reversed-phase liquid chromatography proteomics analysis workflow coupled to nanoelectrospray mass spectrometry for the target analysis of Glu-1-Fibrinopeptide B spiked in a protein digest mixture of bovine serum albumin.
Micromachines [Micromachines (Basel)] 2020 Jan 10; Vol. 11 (1). Date of Electronic Publication: 2020 Jan 10.
Microfluidic devices are gaining increasing popularity due to their wide applications in various research areas. Herein, we propose a two-layer multi-channel microfluidic device allowing for direct-contact cell-vessel co-culture. Using the device, we built a co-culture model of the outer blood-retina barrier (oBRB), mimicking the in vivo retinal pigment epithelial cells-Bruch membrane-fenestrated choroids. To demonstrate the versatility of the design, we further modified the device by inserting platinum electrodes for trans-epithelial electrical resistance (TEER) measurement, demonstrating the feasibility of on-chip assessment of the epithelial barrier integrity. Our proposed design allows for direct-contact co-culture of cell-cell or cell-vessel, modifiable for real-time evaluation of the state of the epithelial monolayers.