ABSTRACT
This article unveiled the development of an inexpensive, lightweight, easy-to-use, and portable fluorescence imaging device for paper-based analytical applications. We used commercial fluorescent dyes, as proof of concept, to verify the feasibility of our fluorescence imaging device for bioanalysis. This approach may provide an alternative method for nucleotide detection and semen analysis, using a miniaturized fluorescence reader that is more compact and portable than conventional analytical equipment.
ABSTRACT
This paper describes a simple design for increasing the tolerance of reagent dislocation on a paper-based platform using a combination of wax-treated paper and a vortex mixer. To date, massive budgetary funds are required in the biotechnological industry to develop new applications; a large part of that cost is attributable to the screening of specific chemical compounds. Here, we propose using a liquid-handling robot to automatically deposit selected reagents on a paper-based platform. We also present a preliminary concept approach for developing a reagent placing device with simple and inexpensive features. A defect of inaccuracy was observed between droplet location and test well location after viewing the performance of the liquid-handling robot on our paper-based platform. Because of dislocation error resulting from robotic reagent placement, we decided to apply an external, rotational force following droplet placement in order to compensate for the distance of reagent dislocation. Note, the largest distance of reagent dislocation was determined by examining the results of altering applied reagent volume, but not concentration, in volumes from 5 µL to 30 µL in a series of experiments. As a result of these experiments, we observed that dislocation was positively affected by an increase in applied volume. A colorimetric assay for nitrite detection was also performed to confirm the feasibility of this method. This work, we believe, can minimize the cost of chemical compound screening for the biotechnological industry.
Subject(s)
Chemistry Techniques, Analytical/instrumentation , Paper , Chemistry Techniques, Analytical/economics , Colorimetry , Hydrophobic and Hydrophilic Interactions , Nitrites/chemistry , RoboticsABSTRACT
This review describes the microfluidic techniques developed for the analysis of a single cell. The characteristics of microfluidic (e.g., little sample amount required, high-throughput performance) make this tool suitable to answer and to solve biological questions of interest about a single cell. This review aims to introduce microfluidic related techniques for the isolation, trapping and manipulation of a single cell. The major approaches for detection in single-cell analysis are introduced; the applications of single-cell analysis are then summarized. The review concludes with discussions of the future directions and opportunities of microfluidic systems applied in analysis of a single cell.
Subject(s)
Microfluidics/methods , Single-Cell Analysis/methods , Animals , Cell Separation , Humans , Lab-On-A-Chip DevicesABSTRACT
This paper describes the development of nucleic acid detection in paper using a combination of commercial fluorescent probes and DNA ladders, and provides us with a better understanding of the interactions between double-stranded DNA (the amplified products in this study), fiber structures in paper, and fluorescent probes. The amplified products (the reverse-transcription and amplification of dengue virus serotype-2 RNA via RT-LAMP) in this study were subsequently fluorescently labeled in paper-based test zones (on our paper-based diagnostic device), thus fluorescent probes were used to perform the diagnosis of dengue fever, specific to serotype-2.
Subject(s)
Dengue Virus/isolation & purification , Dengue/virology , Molecular Diagnostic Techniques/methods , Nucleic Acid Amplification Techniques/methods , Paper , DNA, Viral/genetics , DNA, Viral/isolation & purification , Dengue Virus/classification , Dengue Virus/genetics , Equipment Design , Fluorescent Dyes/chemistry , Humans , Image Processing, Computer-Assisted , Molecular Diagnostic Techniques/instrumentation , Nucleic Acid Amplification Techniques/instrumentation , Oligonucleotide Probes/genetics , RNA, Viral/genetics , RNA, Viral/isolation & purification , Reverse Transcriptase Polymerase Chain Reaction/instrumentation , Reverse Transcriptase Polymerase Chain Reaction/methods , Sensitivity and SpecificityABSTRACT
The development of optofluidic-based technology has ushered in a new era of lab-on-a-chip functionality, including miniaturization of biomedical devices, enhanced sensitivity for molecular detection, and multiplexing of optical measurements. While having great potential, optofluidic devices have only begun to be exploited in many biotechnological applications. Here, we highlight the potential of integrating optofluidic devices with synthetic biological systems, which is a field focusing on creating novel cellular systems by engineering synthetic gene and protein networks. First, we review the development of synthetic biology at different length scales, ranging from single-molecule, single-cell, to cellular population. We emphasize light-sensitive synthetic biological systems that would be relevant for the integration with optofluidic devices. Next, we propose several areas for potential applications of optofluidics in synthetic biology. The integration of optofluidics and synthetic biology would have a broad impact on point-of-care diagnostics and biotechnology.
Subject(s)
Microfluidic Analytical Techniques/methods , Optics and Photonics/methods , Bacteria/metabolism , Fluorescence Resonance Energy Transfer , Fluorescent Dyes/chemistry , Microfluidic Analytical Techniques/instrumentation , Miniaturization , Optics and Photonics/instrumentation , Proteins/metabolism , RNA/metabolism , Synthetic BiologyABSTRACT
Most animals exhibit innate auditory behaviors driven by genetically hardwired neural circuits. In Drosophila, acoustic information is relayed by Johnston organ neurons from the antenna to the antennal mechanosensory and motor center (AMMC) in the brain. Here, by using structural connectivity analysis, we identified five distinct types of auditory projection neurons (PNs) interconnecting the AMMC, inferior ventrolateral protocerebrum (IVLP), and ventrolateral protocerebrum (VLP) regions of the central brain. These auditory PNs are also functionally distinct; AMMC-B1a, AMMC-B1b, and AMMC-A2 neurons differ in their responses to sound (i.e., they are narrowly tuned or broadly tuned); one type of audioresponsive IVLP commissural PN connecting the two hemispheres is GABAergic; and one type of IVLP-VLP PN acts as a generalist responding to all tested audio frequencies. Our findings delineate an auditory processing pathway involving AMMCâIVLPâVLP in the Drosophila brain.