Journal: Micromachines

Article Title: Module-Fluidics: Building Blocks for Spatio-Temporal Microenvironment Control

doi: 10.3390/mi13050774

Figure Lengend Snippet: The Module-Fluidic generates the desired concentration profile that enables downstream microfluidic devices’ function.

Article Snippet: All the valves are controlled by a microfluidic valve control matrix (MUX QUAKE VALVE, Elveflow) with sixteen independent pressure outlets that can provide a constant pressure input when they are switched on.

Techniques: Concentration Assay

Journal: Micromachines

Article Title: Module-Fluidics: Building Blocks for Spatio-Temporal Microenvironment Control

doi: 10.3390/mi13050774

Figure Lengend Snippet: Structures of the Spatio-Temporal Concentration Controller. ( A ) 3D structure of Signal generator (Oscillator) and Module Connector (Integrator), the Oscillator consists of three layers, the PDMS structure layer has flow channels patterned. The channel is molded using positive photo resist which produces a semi-cylindrical cross-section for complete sealing of the micro valve. The second layer is a thin PDMS membrane with controlling valves when the air pressure (∼20 psi) is applied, valve will seal the channel. The third layer is glass slide to support the entire structure. All layers are bounded to each other using plasma bonding. The Integrator is a single microfluidic channel with depth 25 µm. The PDMS layer is also bounded to a glass slide; ( B ) Drawings of the oscillator and integrator ( C ) Concentration signal generation achieved by combination of valve status; ( D ) Function generated by the oscillator.

Article Snippet: All the valves are controlled by a microfluidic valve control matrix (MUX QUAKE VALVE, Elveflow) with sixteen independent pressure outlets that can provide a constant pressure input when they are switched on.

Techniques: Concentration Assay, Membrane, Clinical Proteomics, Generated

Journal: International journal of molecular sciences

Article Title: A Dual-Function "TRE-Lox" System for Genetic Deletion or Reversible, Titratable, and Near-Complete Downregulation of Cathepsin D.

doi: 10.3390/ijms24076745

Figure Lengend Snippet: Figure 1. Overview of the design and dual-functionality of the TRE-Lox system. (a) Overall structure of the 5′ end of the murine cathepsin D (CatD) gene (CTSD) and its promoter region (PCTSD, dark gray), indicating the relative position of the two gRNAs (black arrows) used for CRISPR/Cas9- assisted homologous recombination. Note the placement of the TATA box (TATA) very close to the main transcription start site (TSS) (right-angle arrow), the presence of the initiation codon (ATG, dashed white line) within Exon 1 (Ex 1, light gray), and the presence of a splice donor (SD) and splice acceptor (SA) flanking Intron 1 (black line). (b) Structure of the TRE-Lox knock-in (KI) insert, illustrating the relative positions of the two tet-operons (tetO2, green) and one LoxP site (LoxP, light blue) within the 5′ untranslated region (5′UTR) and, within Intron 1, a tetracycline response element (TRE) comprised of seven tetO repeats (tetO7) and the second LoxP site. The relative placement of the puromycin resistance cassette (Puror, purple) flanked by two FRT sites (FRT, dark blue), which is excisable by Flp recombinase, is depicted using a curly bracket. (c) Downregulation of CTSD via the action of rtTRKRAB acting on the TRE-Lox insert. In the presence of Dox (red triangles), rtTRKRAB binds to the tetO repeats within both the 5′UTR and Intron 1, triggering methylation of histones in a radius of 2–3 kb, thereby remodeling the chromatin and silencing the CTSD gene. (d) Genetic deletion of CTSD via the action of Cre recombinase on the TRE-Lox insert. The figure depicts the end result of Cre-mediated recombination of the TRE-Lox KI insert, which causes removal of the initiation codon, the first portion of the coding region of Exon 1 encoding the signal peptide of CatD, and the 5′ end of Intron 1.

Article Snippet: Relevant regions (and their sources) were as follows: the 3′ end of Exon 1 and the 5′ portion of Intron 1 of murine CTSD (from C57Bl6/J mouse tail DNA); tetO2 (from Addgene plasmid #113892 [41]); TRE 3G and, separately, an FRT-flanked puromycin resistance cassette (both from Addgene plasmid #156430 [42]).

Techniques: CRISPR, Homologous Recombination, Knock-In, Methylation