scholarly journals Development of a Low-Cost, Wireless Smart Thermostat for Isothermal DNA Amplification in Lab-On-A-Chip Devices

Micromachines ◽  
2019 ◽  
Vol 10 (7) ◽  
pp. 437
Author(s):  
Pardy ◽  
Sink ◽  
Koel ◽  
Rang
Keyword(s):  
Dna Analysis ◽  
Low Cost ◽  
Lab On A Chip ◽  
Dna Amplification ◽  
Nucleic Acid Amplification ◽  
Battery Life ◽  
Worst Case ◽  
3D Printed ◽  
Living Organisms ◽  
Smart Thermostat

Nucleic acid amplification tests (NAAT) are widely used for the detection of living organisms, recently applied in Lab-on-a-Chip (LoC) devices to make portable DNA analysis platforms. While portable LoC-NAAT can provide definitive test results on the spot, it requires specialized temperature control equipment. This work focuses on delivering a generalized low-cost, wireless smart thermostat for isothermal NAAT protocols in 2 cm × 3 cm LoC cartridges. We report on the design, prototyping, and evaluation results of our smart thermostat. The thermostat was evaluated by experimental and simulated thermal analysis using 3D printed LoC cartridges, in order to verify its applicability to various isothermal NAAT protocols. Furthermore, it was tested at the boundaries of its operating ambient temperature range as well as its battery life was evaluated. The prototype thermostat was proven functional in 20–30 °C ambient range, capable of maintaining the required reaction temperature of 12 isothermal NAAT protocols with 0.7 °C steady-state error in the worst case.

A Simple Paper-Based Lab-on-a-Chip for the Detection of a Highly Pathogenic Strain of Porcine Reproductive and Respiratory Syndrome Virus

2014 ◽  
Vol 67 (10) ◽  
pp. 1434 ◽  
Author(s):  
Piyasak Chaumpluk ◽  
Annop Suriyasomboon
Keyword(s):  
Signal Detection ◽  
Limit Of Detection ◽  
Low Cost ◽  
Dna Amplification ◽  
High Specificity ◽  
Visual Observation ◽  
Nucleic Acid Amplification ◽  
Pathogenic Strain ◽  
Respiratory Syndrome Virus ◽  
Highly Pathogenic

A paper-based laboratory-on-a-chip assay for the rapid detection of a highly pathogenic strain of porcine reproductive and respiratory syndrome virus (HP-PRRSV) was developed for the first time. The single-unit chip was simply fabricated using Whatman filter paper and plastic lamination. The chip measured 2.5 × 3.0 cm2 and was divided into two parts, one for nucleic acid amplification and the other for signal detection. The HP-PRRSV assay was performed by specific ORF I Nsp 2 gene amplification via an isothermal reverse transcription loop-mediated DNA amplification platform, whereas the cDNA signal detection was performed by visual observation of colorimetric changes in blue silver nanoplates (AgNPls). Positive results caused non-aggregation of the blue AgNPls on the detection pad, whereas negative results induced colorimetric changes in the AgNPls from blue to colourless on the pad. The assay had a limit of detection of 100 copies of the target Nsp 2 gene and high specificity for other types of infectious viruses. The assay required only one hour to complete. This work demonstrates a simple and rapid assay for viruses using a simple, low-cost, paper-based chip.

scholarly journals Highly performing point-of-care molecular testing for SARS-CoV-2 with RNA extraction and isothermal amplification.

2020 ◽  
Author(s):  
Pierre Garneret ◽  
Etienne Coz ◽  
Elian Martin ◽  
Jean-Claude Manuguerra ◽  
Elodie Brient-Litzler ◽  
...  
Keyword(s):  
Point Of Care ◽  
Low Cost ◽  
Rna Extraction ◽  
Dna Amplification ◽  
Developed Countries ◽  
Temperature Cycling ◽  
Isothermal Amplification ◽  
Nucleic Acid Amplification ◽  
Clinical Samples ◽  
Molecular Testing

In order to respond to the urgent request of massive testing, developed countries perform nucleic acid amplification tests (NAAT) of SARS-CoV-2 in centralized laboratories. Real-time RT - PCR (Reverse transcription - Polymerase Chain Reaction) is used to amplify the viral RNA and enable its detection. Although PCR is 37 years old, it is still considered, without dispute, as the gold standard. PCR is an efficient process, but the complex engineering required for automated RNA extraction and temperature cycling makes it incompatible for use in point of care settings. In the present work, by harnessing progress made in the past two decades in DNA amplification, microfluidics and membrane technologies, we succeeded to create a portable test, in which SARS-CoV-2 RNA is extracted, amplified isothermally by RT - LAMP (Loop-mediated Isothermal Amplification), and detected using intercalating dyes or highly fluorescent probes. Depending on the viral load, the detection takes between twenty minutes and one hour. Using pools of naso-pharyngal clinical samples, we estimated a sensitivity comparable to RT-qPCR (up to a Cycle threshold of 39, equivalent to <0.1 TCID50 per mL) and a 100% specificity, for other human coronaviruses and eight respiratory viruses currently circulating in Europe. We designed and fabricated an easy-to-use portable device called COVIDISC to carry out the test at the point of care. The low cost of the materials along with the absence of complex equipment paves the way towards a large dissemination of this device. The perspective of a reliable SARS-CoV-2 point of care detection, highly performing, that would deliver on-site results in less than one hour opens up a new efficient approach to manage the pandemics.

scholarly journals Bioluminescent detection of isothermal DNA amplification in microfluidic generated droplets and artificial cells

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Patrick Hardinge ◽  
Divesh K. Baxani ◽  
Thomas McCloy ◽  
James A. H. Murray ◽  
Oliver K. Castell
Keyword(s):  
Real Time ◽  
High Throughput ◽  
Soft Matter ◽  
Low Cost ◽  
Diagnostic Technique ◽  
Dna Amplification ◽  
Droplet Microfluidics ◽  
Nucleic Acid Amplification ◽  
Molecular Diagnostic ◽  
Artificial Cells

AbstractMicrofluidic droplet generation affords precise, low volume, high throughput opportunities for molecular diagnostics. Isothermal DNA amplification with bioluminescent detection is a fast, low-cost, highly specific molecular diagnostic technique that is triggerable by temperature. Combining loop-mediated isothermal nucleic acid amplification (LAMP) and bioluminescent assay in real time (BART), with droplet microfluidics, should enable high-throughput, low copy, sequence-specific DNA detection by simple light emission. Stable, uniform LAMP–BART droplets are generated with low cost equipment. The composition and scale of these droplets are controllable and the bioluminescent output during DNA amplification can be imaged and quantified. Furthermore these droplets are readily incorporated into encapsulated droplet interface bilayers (eDIBs), or artificial cells, and the bioluminescence tracked in real time for accurate quantification off chip. Microfluidic LAMP–BART droplets with high stability and uniformity of scale coupled with high throughput and low cost generation are suited to digital DNA quantification at low template concentrations and volumes, where multiple measurement partitions are required. The triggerable reaction in the core of eDIBs can be used to study the interrelationship of the droplets with the environment and also used for more complex chemical processing via a self-contained network of droplets, paving the way for smart soft-matter diagnostics.

scholarly journals Portable Molecular Diagnostics Device for Identification of Asini Corii Colla by Loop-Mediated Isothermal Amplification

Inventions ◽  
2021 ◽  
Vol 6 (3) ◽  
pp. 51
Author(s):  
Shyangchwen Sheu ◽  
Chungyu Huang ◽  
Jyhjian Chen
Keyword(s):  
Dna Analysis ◽  
Limit Of Detection ◽  
Low Cost ◽  
Dna Amplification ◽  
Thermal Control ◽  
Rechargeable Batteries ◽  
Isothermal Amplification ◽  
Portable Device ◽  
Chinese Medicines ◽  
Loop Mediated Isothermal Amplification

Asini Corii Colla (ACC; donkey-hide glue) is one of the most valuable tonic traditional Chinese medicines. Because of the large demand for gelatinous Chinese medicines, bovine or swine skin was sometimes used to make adulterated gelatine in recent decades. Food chemicals can greatly harm people’s health, and detecting chemicals in foods is extremely important. A loop-mediated isothermal amplification (LAMP) device with smartphone detection is demonstrated in this study for detecting the DNA of Asini Corii Colla. The complete system is composed of a hand-held box equipped with a smartphone, a cartridge heater, an ultraviolet LED, a disposable reaction tube, and a homemade thermal module. All the processes are powered by a set of rechargeable batteries. Comprehensive experiments of measuring temperature profiles are presented, which showed the accuracy of temperature under thermal control is less than 0.5 °C. By implementing one heating module with an ATmega328p-au microcontroller in the device, the DNA mixture is heated directly up to the reaction temperature within 5 min. Next, a DNA segment of Asini Corii Colla is utilized to evaluate the sensitivity of the DNA amplification in the portable device. A limit of detection to a concentration of 10−4 ng/μL is achieved. Real-time detection of Asini Corii Colla by a smartphone camera can be achieved using this portable device. The unique architecture utilized in this device is ideal for a low-cost DNA analysis system.

scholarly journals 3D Printed Lab-on-a-Chip Platform for Chemical Stimulation and Parallel Analysis of Ion Channel Function

Micromachines ◽  
2019 ◽  
Vol 10 (8) ◽  
pp. 548 ◽  
Author(s):  
Daniel Aschenbrenner ◽  
Oliver Friedrich ◽  
Daniel F. Gilbert
Keyword(s):  
Ion Channels ◽  
Ion Channel ◽  
Low Cost ◽  
Lab On A Chip ◽  
Chemical Stimulation ◽  
Hek293 Cells ◽  
Solution Exchange ◽  
3D Printed

Functional imaging has been a widely established method for the assessment of ion channel function in vitro. Conventional infrastructure used for in vitro functional analysis of ion channels is typically proprietary, non-customizable, expensive, and requires a high level of skill to use and maintain. 3D desktop printing, which is employed in the rapid prototyping field, allows for quick engineering of alternatives to conventional imaging infrastructure that are customizable, low cost, and user friendly. Here, we describe an ultra-low-cost microfluidic lab-on-a-chip (LOC) device manufactured using acrylonitrile butadiene styrene (ABS) for in vitro functional imaging of ion channels that can quickly and easily be reconstructed using three-dimensional (3D) desktop printing. The device is light weight (<5 g), small (20 mm × 49 mm), and extremely low cost (<EUR 1). We simulate fluidics within the printed channels and assess the suitability of the engineered chamber to generate homogeneous mixtures during solution exchange. We demonstrate the usability of the 3D printed microfluidic device in a case study using Fluo-4-loaded human embryonal kidney-derived (HEK293) cells, recombinantly expressing the capsaicin receptor, transient receptor potential vanilloid receptor type 1 (TRPV1), as a model system. In the case study, we confirm its applicability to solution exchange for chemical stimulation and parallel functional time-lapse fluorescence microscopy-based calcium imaging. We assess the suitability of ABS for culturing HEK293 cells inside the microfluidic LOC, based on qualitative analysis of microscopic transmission light images of ABS-exposed HEK293 cells and confirm the previously reported biocompatibility of ABS. To highlight the versatility of the 3D printed microfluidic device, we provide an example for multiplication of the shown concept within a 3D printed multichannel microfluidic LOC to be used, for example, in a higher throughput format for parallelized functional analysis of ion channels. While this work focusses on Ca2+ imaging with TRPV1 channels, the device may also be useful for application with other ion channel types and in vitro models.

Low cost lab-on-a-chip prototyping with a consumer grade 3D printer

Lab on a Chip ◽  
2014 ◽  
Vol 14 (16) ◽  
pp. 2978-2982 ◽  
Author(s):  
Germán Comina ◽  
Anke Suska ◽  
Daniel Filippini
Keyword(s):  
Low Cost ◽  
Lab On A Chip ◽  
3D Printer ◽  
3D Printed

Versatile prototyping of 3D printed lab-on-a-chip devices, supporting different forms of sample delivery, transport, functionalization and readout, is demonstrated with a consumer grade printer, which centralizes all critical fabrication tasks.

Pioneering Low-Cost 3D-Printed Transtibial Prosthetics to Serve a Rural Population in Sierra Leone

2020 ◽  
Author(s):  
Merel van der Stelt ◽  
Martin P. Grobusch ◽  
Abdul R. Koroma ◽  
Marco Papenburg ◽  
Ismaila Kebbie ◽  
...  
Keyword(s):  
Sierra Leone ◽  
Rural Population ◽  
Low Cost ◽  
3D Printed

scholarly journals 3D printed microfluidic lab-on-a-chip device for fiber-based dual beam optical manipulation

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Haoran Wang ◽  
Anton Enders ◽  
John-Alexander Preuss ◽  
Janina Bahnemann ◽  
Alexander Heisterkamp ◽  
...  
Keyword(s):  
Optical Fibers ◽  
Single Cells ◽  
Lab On A Chip ◽  
Cost Effective ◽  
Optical Manipulation ◽  
Hydrodynamic Focusing ◽  
Trapping Region ◽  
Dual Beam ◽  
3D Printed ◽  
On Chip

Abstract3D printing of microfluidic lab-on-a-chip devices enables rapid prototyping of robust and complex structures. In this work, we designed and fabricated a 3D printed lab-on-a-chip device for fiber-based dual beam optical manipulation. The final 3D printed chip offers three key features, such as (1) an optimized fiber channel design for precise alignment of optical fibers, (2) an optically clear window to visualize the trapping region, and (3) a sample channel which facilitates hydrodynamic focusing of samples. A square zig–zag structure incorporated in the sample channel increases the number of particles at the trapping site and focuses the cells and particles during experiments when operating the chip at low Reynolds number. To evaluate the performance of the device for optical manipulation, we implemented on-chip, fiber-based optical trapping of different-sized microscopic particles and performed trap stiffness measurements. In addition, optical stretching of MCF-7 cells was successfully accomplished for the purpose of studying the effects of a cytochalasin metabolite, pyrichalasin H, on cell elasticity. We observed distinct changes in the deformability of single cells treated with pyrichalasin H compared to untreated cells. These results demonstrate that 3D printed microfluidic lab-on-a-chip devices offer a cost-effective and customizable platform for applications in optical manipulation.

scholarly journals UV Inscription and Pressure Induced Long-Period Gratings through 3D Printed Amplitude Masks

Sensors ◽  
2021 ◽  
Vol 21 (6) ◽  
pp. 1977
Author(s):  
Ricardo Oliveira ◽  
Liliana M. Sousa ◽  
Ana M. Rocha ◽  
Rogério Nogueira ◽  
Lúcia Bilro
Keyword(s):  
State Of The Art ◽  
Low Cost ◽  
Resonance Wavelength ◽  
Complex Structures ◽  
Pressing Method ◽  
Long Period ◽  
Long Period Gratings ◽  
3D Printed ◽  
Mechanical Pressing ◽  
First Time

In this work, we demonstrate for the first time the capability to inscribe long-period gratings (LPGs) with UV radiation using simple and low cost amplitude masks fabricated with a consumer grade 3D printer. The spectrum obtained for a grating with 690 µm period and 38 mm length presented good quality, showing sharp resonances (i.e., 3 dB bandwidth < 3 nm), low out-of-band loss (~0.2 dB), and dip losses up to 18 dB. Furthermore, the capability to select the resonance wavelength has been demonstrated using different amplitude mask periods. The customization of the masks makes it possible to fabricate gratings with complex structures. Additionally, the simplicity in 3D printing an amplitude mask solves the problem of the lack of amplitude masks on the market and avoids the use of high resolution motorized stages, as is the case of the point-by-point technique. Finally, the 3D printed masks were also used to induce LPGs using the mechanical pressing method. Due to the better resolution of these masks compared to ones described on the state of the art, we were able to induce gratings with higher quality, such as low out-of-band loss (0.6 dB), reduced spectral ripples, and narrow bandwidths (~3 nm).

scholarly journals BoSL FAL Pump: a small, low-cost, easily constructed, 3D-printed peristaltic pump for sampling of waters

HardwareX ◽  
2021 ◽  
pp. e00214
Author(s):  
David T. McCarthy ◽  
Baiqian Shi ◽  
Miao Wang ◽  
Stephen Catsamas
Keyword(s):  
Low Cost ◽  
Peristaltic Pump ◽  
3D Printed
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