Digital Lamp Research Articles

A microfluidic chip capable of generating and trapping emulsion droplets for digital loop-mediated isothermal amplification analysis.

Loop-mediated isothermal amplification (LAMP) is a nucleic acid amplification technique that rapidly amplifies specific DNA molecules at high yield. In this study, a microfluidic droplet array chip was designed to execute the digital LAMP process. The novel device was capable of 1) creating emulsion droplets, 2) sorting them into a 30 × 8 droplet array, and 3) executing LAMP across the 240 trapped and separated droplets (with a volume of 0.22 nL) after only 40 min of reaction at 56 °C. Nucleic acids were accurately quantified across a dynamic range of 50 to 2.5 × 103 DNA copies per μL, and the limit of detection was a single DNA molecule. This is the first time that an arrayed emulsion droplet microfluidic device has been used for digital LAMP analysis. When compared to microwell digital nucleic acid amplification assays, this droplet array-based digital LAMP assay eliminates the constraint on the size of the digitized target, which was determined by the dimension of the microwells for its counterparts. Moreover, the capacity for hydrodynamic droplet trapping allows the chip to operate in a one-droplet-to-one-trap manner. This microfluidic chip may therefore become a promising device for digital LAMP-based diagnostics in the near future.

Rapid pathogen-specific phenotypic antibiotic susceptibility testing using digital LAMP quantification in clinical samples.

Rapid antimicrobial susceptibility testing (AST) is urgently needed for informing treatment decisions and preventing the spread of antimicrobial resistance resulting from the misuse and overuse of antibiotics. To date, no phenotypic AST exists that can be performed within a single patient visit (30 min) directly from clinical samples. We show that AST results can be obtained by using digital nucleic acid quantification to measure the phenotypic response of Escherichia coli present within clinical urine samples exposed to an antibiotic for 15 min. We performed this rapid AST using our ultrafast (~7 min) digital real-time loop-mediated isothermal amplification (dLAMP) assay [area under the curve (AUC), 0.96] and compared the results to a commercial (~2 hours) digital polymerase chain reaction assay (AUC, 0.98). The rapid dLAMP assay can be used with SlipChip microfluidic devices to determine the phenotypic antibiotic susceptibility of E. coli directly from clinical urine samples in less than 30 min. With further development for additional pathogens, antibiotics, and sample types, rapid digital AST (dAST) could enable rapid clinical decision-making, improve management of infectious diseases, and facilitate antimicrobial stewardship.

Simple Digital D1S Lamp System with a Power Structure of Flyback Converter and Full-Bridge Inverter

Simple Digital D1S Lamp System with a Power Structure of Flyback Converter and Full-Bridge Inverter

Digital droplet LAMP as a microfluidic app on standard laboratory devices

Digital droplet LAMP is performed on a disposable chip (DropChip) with the size of a microscope slide using only standard laboratory devices.

Comparative Study of Sensitivity, Linearity, and Resistance to Inhibition of Digital and Nondigital Polymerase Chain Reaction and Loop Mediated Isothermal Amplification Assays for Quantification of Human Cytomegalovirus

Performing nucleic acid amplification techniques (NAATs) in digital format using limiting dilution provides potential advantages that have recently been demonstrated with digital polymerase chain reaction (dPCR). Key benefits that have been claimed are the ability to quantify nucleic acids without the need of an external calibrator and a greater resistance to inhibitors than real-time quantitative PCR (qPCR). In this study, we evaluated the performance of four NAATs, qPCR, dPCR, real-time quantitative loop mediated isothermal amplification (qLAMP), and digital LAMP (dLAMP), for the detection and quantification of human cytomegalovirus (hCMV). We used various DNA templates and inhibitors to compare the performance of these methods using a conventional real-time thermocycler platform (Bio-Rad CFX96) and a chip based digital platform (Fluidigm Biomark 12.765 Digital Array). dPCR performed well and demonstrated greater resistance to inhibitors than the other methods although this resistance did not apply equally to all inhibitors tested. dLAMP was found to be less sensitive than dPCR, but its quantitative performance was better than qLAMP, the latter being unable to quantify below 1000 copies. dLAMP was also more resistant to inhibitors than qLAMP. Unlike qPCR, both digital methods were able to quantify viral genomes without requiring a calibrator; however, neither can currently compete with the large reaction volumes, and thus the greater absolute sensitivity, of qPCR. With the introduction of digital instrumentation that will enable larger reaction volumes, digital amplification methods such as those evaluated in this study could potentially offer a robust alternative to qPCR for nucleic acid quantification.

Digital Lamp Balance System With Individual Lamp Current Control Capability for an LCD TV

A digital lamp balance system that equalizes lamp current of liquid crystal display (LCD) backlight is implemented in this letter. The proposed digital balance circuit consists of a balance switch which is in series with the lamp and a dc-blocking capacitor which also acts as a ballast capacitor for parallel lighting. The parasitic capacitance of the balance switch is used as parallel impedance which makes the lamp current decrease. Microcontroller unit controls each lamp current by changing the pulsewidth modulation duty ratio of a balance switch. The specified circuit, analysis, balance principles, and experimental results of the proposed system will be presented for a 46-in LCD TV.

Digital LAMP in a sample self-digitization (SD) chip

This paper describes the realization of digital loop-mediated DNA amplification (dLAMP) in a sample self-digitization (SD) chip. Digital DNA amplification has become an attractive technique to quantify absolute concentrations of DNA in a sample. While digital polymerase chain reaction is still the most widespread implementation, its use in resource-limited settings is impeded by the need for thermal cycling and robust temperature control. In such situations, isothermal protocols that can amplify DNA or RNA without thermal cycling are of great interest. Here, we accomplished the successful amplification of single DNA molecules in a stationary droplet array using isothermal digital loop-mediated DNA amplification. Unlike most (if not all) existing methods for sample discretization, our design allows for automated, loss-less digitization of sample volumes on-chip. We demonstrated accurate quantification of relative and absolute DNA concentrations with sample volumes of less than 2 μl. We assessed the homogeneity of droplet size during sample self-digitization in our device, and verified that the size variation was small enough such that straightforward counting of LAMP-active droplets sufficed for data analysis. We anticipate that the simplicity and robustness of our SD chip make it attractive as an inexpensive and easy-to-operate device for DNA amplification, for example in point-of-care settings.

Restricted post-trabeculectomy bleb formation by conjunctival scarring

To evaluate the effects of radial conjunctival incision during fornix-based trabeculectomy, and conjunctival scarring associated with previous ocular surgeries, for the restriction of bleb formation. Digital photo-slit lamp images of filtering blebs, which had undergone fornix-based trabeculectomy with mitomycin C previously, were analysed. The area and the height of the bleb in the conjunctivae with radial incision (Ain and Hin, respectively) were compared with the area and the height of the bleb in the conjunctivae without radial incision (Ano and Hno, respectively). We compared the parameters of bleb area (total bleb area, Ain and Ano) and bleb height (the bleb height at the centre of the scleral flap; Hc, Hin and Hno) in eyes without previous ocular surgeries, with those in eyes that underwent previous ocular surgeries. The study population consisted of 51 eyes. The Ano and the Hno were significantly larger than the Ain (p < 0.001) and the Hin (p < 0.001) respectively. Subgroup analyses in eyes without previous ocular surgeries and eyes with phacoemulsification cataract surgery also showed that the Ano was significantly larger than the Ain. However, there was no significant difference between the Ano and the Ain in eyes with previous trabeculectomy. A comparison of bleb formation between eyes without previous ocular surgeries and eyes with cataract surgery showed no significant differences in each of the bleb area and bleb height parameters. By contrast, the total bleb area (p = 0.021), the Ano (p = 0.017) and the Hc (p = 0.045) were significantly smaller in eyes with previous trabeculectomy than in eyes without previous ocular surgeries. Our data demonstrate that bleb formation depends on radial conjunctival incision during fornix-based trabeculectomy and surgical conjunctival scarring from previous trabeculectomy.

Electronic Ballast Control IC With Digital Phase Control and Lamp Current Regulation

A digital control architecture is presented for electronic ballasts that provides a phase sweep for reliable, soft lamp ignition and a smooth transition to lamp current regulation mode. The controller is based on an inner phase loop for fast regulation of the resonant tank operating point and an outer current loop for lamp current regulation. The inner loop operates on a simple digital control law that computes the required gate timing relative to the inductor current positive zero crossing. Phase control provides reliable drive of the resonant converter in the presence of large dynamic changes in the load impedance during lamp ignition and warm up and natural tracking of component variations with temperature and time. The primarily digital approach provides programmability for broad application, insensitivity to process and temperature variations, realization in low cost CMOS processes and few external components. Experimental results are presented for an integrated ballast controller fabricated in a 0.8 mum CMOS process used in a 400 W, 150 kHz HID electronic ballast.

Bovine serum albumin reverses inhibition of RT-PCR by melanin.

Bovine serum albumin reverses inhibition of RT-PCR by melanin.

Instructional display device operated responsive to the playing of stringed musical instruments

Separate areas of a display board are selectively activated either separately or in various combinations to illustrate a chord diagram, a fingerboard diagram, and/or a musical staff diagram. Lamps in each of the three diagrams are illuminated responsive to the playing of a specially constructed stringed instrument by the instructor to illustrate finger positions in the case of the chord diagram and the fingerboard diagram, and to illustrate musical position of the notes played on the scale diagram. One or more lock-in switches cause the activated lamps to remain lighted after the strings of the musical instrument are released by the instructor. Finally a digital readout lamp associated with each string on the chord diagram illustrates the finger which should be used to depress the corresponding string at the location indicated by the lamp.