Analysis of Existing Thermodynamic Models of the Liquid Drop Deposited on the Substrate—A Sufficient Condition of the Minimum Free Energy of the System

On the basis of the principles of non-equilibrium thermodynamics, the following condition was determined: necessary and sufficient for the occurrence of a minimum free energy of a liquid droplet deposited on a solid substrate in a gaseous environment in an isothermal and isochoric system. Only for positive values of the energy of three-phase tension line (shrinking the wetting circumference) for small and large contact angles can the system not reach this minimum. Without exceeding a certain free energy limit, it is not possible for the drop to spontaneously spread over the surface. For zero and negative energy of three-phase tension line (stretching the wetting circumference), the system can always reach a minimum of free energy. The developed equations allow determining the change of free energy occurring between any two stationary states when the droplet volume and physicochemical parameters characterizing energies at the interfaces are known. For a known set of such parameters, the equations allow determining the trajectory of free energy changes in the system as a function of the contact angle from the moment the drop comes into contact with the substrate. The application of the principles of non-equilibrium thermodynamics makes it possible to treat a real system as one in which the drops do not evaporate. However, the system has to be isothermal

Droplet Microfluidic Technique for the Study of Fermentation

We demonstrate a technique that uses microdroplets for culturing and selecting bacterial cultures in a model biotechnological application. We propose an assay for determination of ethanol concentration that provides increased dynamic range and is compatible with droplet microfluidic screening technologies. The assay comprises two enzymes—alcohol oxidase (AOX) and horseradish peroxidase (HRP)—and a colorimetric readout system of phenol-4-sulfonic acid (PSA) and 4-aminoantipyrine (4-AAP). The microdroplet method provides high repeatability (a relative error of measured ethanol concentration < 5%), high specificity for ethanol, low consumption of reagents and wide dynamic range (1–70 g·L-1) compared to existing assays. We report the use of this method in a screen of ethanol generation efficiency of Zymomonas mobilis (strain 3881) against the concentration of glucose in the culture media

Generation of Nanoliter Droplets on Demand at Hundred-Hz Frequencies

We describe a precision micropump for generation of precisely metered micro-aliquots of liquid at high rates. The use of custom designed piezoelectric valves positioned externally to the microfluidic chip allows for on-demand formation of micro-droplets with online control of their individual volumes from nLs to μLs at frequencies up to 400 Hz. The system offers precision of administering volumes of 1% and of time of emission of <0.5 ms. The use of a piezoelectric actuator provides two distinct vistas for controlling the volume of the droplets—either by digital control of the “open” interval or by analogue tuning of the lumen of the valve. Fast and precise generation of droplets make this system a perfect constituent module for microfluidic high-speed combinatorial screening schemes

An Automated Microfluidic System for the Generation of Droplet Interface Bilayer Networks

Networks of droplets, in which aqueous compartments are separated by lipid bilayers, have shown great potential as a model for biological transmembrane communication. We present a microfluidic system which allows for on-demand generation of droplets that are hydrodynamically locked in a trapping structure. As a result, the system enables the formation of a network of four droplets connected via lipid bilayers and the positions of each droplet in the network can be controlled thanks to automation of microfluidic operations. We perform electrophysiological measurements of ionic currents indicating interactions between nanopores and small molecules to prove the potential of the device in screening of the inhibitors acting on membrane proteins. We also demonstrate, for the first time, a microfluidic droplet interface bilayer (DIB) system in which the testing of inhibitors can be performed without direct contact between the tested sample and the electrodes recording picoampere currents

Optical Biosensing System for the Detection of Survivin mRNA in Colorectal Cancer Cells Using a Graphene Oxide Carrier-Bound Oligonucleotide Molecular Beacon

The anti-apoptotic protein survivin is one of the most promising cancer biomarkers owing to its high expression in human cancers and rare occurrence in normal adult tissues. In this work, we have investigated the role of supramolecular interactions between a graphene oxide (GO) nanosheet nanocarrier and a survivin molecular beacon (SurMB), functionalized by attaching fluorophore Joe and quencher Dabcyl (SurMB-Joe). Molecular dynamics simulations revealed hydrogen bonding of Joe moiety and Dabcyl to GO carriers that considerably increase the SurMB-GO bonding strength. This was confirmed in experimental work by the reduced fluorescence background in the OFF state, thereby increasing the useful analytical signal range for mRNA detection. A new mechanism of hairpin–hairpin interaction of GO@SurMB with target oligonucleotides has been proposed. A low limit of detection, LOD = 16 nM (S/N = 3), has been achieved for complementary tDNA using GO@SurMB-Joe nanocarriers. We have demonstrated an efficient internalization of SurMB-Joe-loaded GO nanocarriers in malignant SW480 cells. The proposed tunability of the bonding strength in the attached motifs for MBs immobilized on nanocarriers, via structural modifications, should be useful in gene delivery systems to enhance the efficacy of gene retention, cell transfection and genomic material survivability in the cellular environment

Continuous Recirculation of Microdroplets in a Closed Loop Tailored for Screening of Bacteria Cultures

Emerging microfluidic technology has introduced new precision controls over reaction conditions. Owing to the small amount of reagents, microfluidics significantly lowers the cost of carrying a single reaction. Moreover, in two-phase systems, each part of a dispersed fluid can be treated as an independent chemical reactor with a volume from femtoliters to microliters, increasing the throughput. In this work, we propose a microfluidic device that provides continuous recirculation of droplets in a closed loop, maintaining low consumption of oil phase, no cross-contamination, stabilized temperature, a constant condition of gas exchange, dynamic feedback control on droplet volume, and a real-time optical characterization of bacterial growth in a droplet. The channels (tubing) and junction cubes are made of Teflon fluorinated ethylene propylene (FEP) to ensure non-wetting conditions and to prevent the formation of biofilm, which is particularly crucial for biological experiments. We show the design and operation of a novel microfluidic loop with the circular motion of microdroplet reactors monitored with optical sensors and precision temperature controls. We have employed the proposed system for long term monitoring of bacterial growth during the antibiotic chloramphenicol treatment. The proposed system can find applications in a broad field of biomedical diagnostics and therapy