Microfluidic device
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Microfluidics refers to the behavior, precise control, and manipulation of
fluids In physics, a fluid is a liquid, gas, or other material that continuously deforms (''flows'') under an applied shear stress, or external force. They have zero shear modulus, or, in simpler terms, are substances which cannot resist any she ...
that are geometrically constrained to a small scale (typically sub-millimeter) at which surface forces dominate volumetric forces. It is a multidisciplinary field that involves
engineering Engineering is the use of scientific principles to design and build machines, structures, and other items, including bridges, tunnels, roads, vehicles, and buildings. The discipline of engineering encompasses a broad range of more speciali ...
,
physics Physics is the natural science that studies matter, its fundamental constituents, its motion and behavior through space and time, and the related entities of energy and force. "Physical science is that department of knowledge which r ...
, chemistry,
biochemistry Biochemistry or biological chemistry is the study of chemical processes within and relating to living organisms. A sub-discipline of both chemistry and biology, biochemistry may be divided into three fields: structural biology, enzymology and ...
, nanotechnology, and
biotechnology Biotechnology is the integration of natural sciences and engineering sciences in order to achieve the application of organisms, cells, parts thereof and molecular analogues for products and services. The term ''biotechnology'' was first used ...
. It has practical applications in the design of systems that process low volumes of fluids to achieve multiplexing, automation, and
high-throughput screening High-throughput screening (HTS) is a method for scientific experimentation especially used in drug discovery and relevant to the fields of biology, materials science and chemistry. Using robotics, data processing/control software, liquid handling ...
. Microfluidics emerged in the beginning of the 1980s and is used in the development of
inkjet Inkjet printing is a type of computer printing that recreates a digital image by propelling droplets of ink onto paper and plastic substrates. Inkjet printers were the most commonly used type of printer in 2008, and range from small inexpens ...
printheads, DNA chips,
lab-on-a-chip A lab-on-a-chip (LOC) is a device that integrates one or several laboratory functions on a single integrated circuit (commonly called a "chip") of only millimeters to a few square centimeters to achieve automation and high-throughput screening. ...
technology, micro-propulsion, and micro-thermal technologies. Typically, micro means one of the following features: * Small volumes (μL, nL, pL, fL) * Small size * Low energy consumption * Microdomain effects Typically microfluidic systems transport, mix, separate, or otherwise process fluids. Various applications rely on passive fluid control using
capillary force Capillary action (sometimes called capillarity, capillary motion, capillary rise, capillary effect, or wicking) is the process of a liquid flowing in a narrow space without the assistance of, or even in opposition to, any external forces l ...
s, in the form of capillary flow modifying elements, akin to flow resistors and flow accelerators. In some applications, external actuation means are additionally used for a directed transport of the media. Examples are rotary drives applying centrifugal forces for the fluid transport on the passive chips. Active microfluidics refers to the defined manipulation of the working fluid by active (micro) components such as
micropump Micropumps are devices that can control and manipulate small fluid volumes. Although any kind of small pump is often referred to as micropump, a more accurate definition restricts this term to pumps with functional dimensions in the micrometer ran ...
s or microvalves. Micropumps supply fluids in a continuous manner or are used for dosing. Microvalves determine the flow direction or the mode of movement of pumped liquids. Often, processes normally carried out in a lab are miniaturised on a single chip, which enhances efficiency and mobility, and reduces sample and reagent volumes.


Microscale behaviour of fluids

The behaviour of fluids at the microscale can differ from "macrofluidic" behaviour in that factors such as surface tension, energy dissipation, and fluidic resistance start to dominate the system. Microfluidics studies how these behaviours change, and how they can be worked around, or exploited for new uses. At small scales (channel size of around 100
nanometers 330px, Different lengths as in respect to the molecular scale. The nanometre (international spelling as used by the International Bureau of Weights and Measures; SI symbol: nm) or nanometer (American and British English spelling differences#-re ...
to 500
micrometers The micrometre ( international spelling as used by the International Bureau of Weights and Measures; SI symbol: μm) or micrometer (American spelling), also commonly known as a micron, is a unit of length in the International System of Unit ...
) some interesting and sometimes unintuitive properties appear. In particular, the Reynolds number (which compares the effect of the momentum of a fluid to the effect of
viscosity The viscosity of a fluid is a measure of its resistance to deformation at a given rate. For liquids, it corresponds to the informal concept of "thickness": for example, syrup has a higher viscosity than water. Viscosity quantifies the inte ...
) can become very low. A key consequence is co-flowing fluids do not necessarily mix in the traditional sense, as flow becomes
laminar Laminar means "flat". Laminar may refer to: Terms in science and engineering: * Laminar electronics or organic electronics, a branch of material sciences dealing with electrically conductive polymers and small molecules * Laminar armour or "band ...
rather than
turbulent In fluid dynamics, turbulence or turbulent flow is fluid motion characterized by chaotic changes in pressure and flow velocity. It is in contrast to a laminar flow, which occurs when a fluid flows in parallel layers, with no disruption between ...
; molecular transport between them must often be through
diffusion Diffusion is the net movement of anything (for example, atoms, ions, molecules, energy) generally from a region of higher concentration to a region of lower concentration. Diffusion is driven by a gradient in Gibbs free energy or chemica ...
. High specificity of chemical and physical properties (concentration, pH, temperature, shear force, etc.) can also be ensured resulting in more uniform reaction conditions and higher grade products in single and multi-step reactions.


Various kinds of microfluidic flows

Microfluidic flows need only be constrained by geometrical length scale - the modalities and methods used to achieve such a geometrical constraint are highly dependent on the targeted application. Traditionally, microfluidic flows have been generated inside closed channels with the channel cross section being in the order of 10 μm x 10 μm. Each of these methods has its own associated techniques to maintain robust fluid flow which have matured over several years.


Open microfluidics

The behavior of fluids and their control in open microchannels was pioneered around 2005 and applied in air-to-liquid sample collection and chromatography. In
open microfluidics Microfluidics refers to the flow of fluid in channels or networks with at least one dimension on the micron scale. In open microfluidics, also referred to as open surface microfluidics or open-space microfluidics, at least one boundary confining th ...
, at least one boundary of the system is removed, exposing the fluid to air or another interface (i.e. liquid). Advantages of open microfluidics include accessibility to the flowing liquid for intervention, larger liquid-gas surface area, and minimized bubble formation. Another advantage of open microfluidics is the ability to integrate open systems with surface-tension driven fluid flow, which eliminates the need for external pumping methods such as peristaltic or syringe pumps. Open microfluidic devices are also easy and inexpensive to fabricate by milling, thermoforming, and hot embossing. In addition, open microfluidics eliminates the need to glue or bond a cover for devices, which could be detrimental to capillary flows. Examples of open microfluidics include open-channel microfluidics, rail-based microfluidics, paper-based, and thread-based microfluidics. Disadvantages to open systems include susceptibility to evaporation, contamination, and limited flow rate.


Continuous-flow microfluidics

Continuous flow microfluidics rely on the control of a steady state liquid flow through narrow channels or porous media predominantly by accelerating or hindering fluid flow in capillary elements. In paper based microfluidics, capillary elements can be achieved through the simple variation of section geometry. In general, the actuation of liquid flow is implemented either by external
pressure Pressure (symbol: ''p'' or ''P'') is the force applied perpendicular to the surface of an object per unit area over which that force is distributed. Gauge pressure (also spelled ''gage'' pressure)The preferred spelling varies by country and e ...
sources, external mechanical pumps, integrated mechanical
micropump Micropumps are devices that can control and manipulate small fluid volumes. Although any kind of small pump is often referred to as micropump, a more accurate definition restricts this term to pumps with functional dimensions in the micrometer ran ...
s, or by combinations of capillary forces and electrokinetic mechanisms. Continuous-flow microfluidic operation is the mainstream approach because it is easy to implement and less sensitive to protein fouling problems. Continuous-flow devices are adequate for many well-defined and simple biochemical applications, and for certain tasks such as chemical separation, but they are less suitable for tasks requiring a high degree of flexibility or fluid manipulations. These closed-channel systems are inherently difficult to integrate and scale because the parameters that govern flow field vary along the flow path making the fluid flow at any one location dependent on the properties of the entire system. Permanently etched microstructures also lead to limited reconfigurability and poor fault tolerance capability. Computer-aided design automation approaches for continuous-flow microfluidics have been proposed in recent years to alleviate the design effort and to solve the scalability problems. Process monitoring capabilities in continuous-flow systems can be achieved with highly sensitive microfluidic flow sensors based on
MEMS Microelectromechanical systems (MEMS), also written as micro-electro-mechanical systems (or microelectronic and microelectromechanical systems) and the related micromechatronics and microsystems constitute the technology of microscopic devices, ...
technology, which offers resolutions down to the nanoliter range.


Droplet-based microfluidics

Droplet-based microfluidics is a subcategory of microfluidics in contrast with continuous microfluidics; droplet-based microfluidics manipulates discrete volumes of fluids in immiscible phases with low Reynolds number and laminar flow regimes. Interest in droplet-based microfluidics systems has been growing substantially in past decades. Microdroplets allow for handling miniature volumes (μl to fl) of fluids conveniently, provide better mixing, encapsulation, sorting, and sensing, and suit high throughput experiments. Exploiting the benefits of droplet-based microfluidics efficiently requires a deep understanding of droplet generation to perform various logical operations such as droplet manipulation, droplet sorting, droplet merging, and droplet breakup.


Digital microfluidics

Alternatives to the above closed-channel continuous-flow systems include novel open structures, where discrete, independently controllable droplets are manipulated on a substrate using
electrowetting Electrowetting is the modification of the wetting properties of a surface (which is typically hydrophobic) with an applied electric field. History The electrowetting of mercury and other liquids on variably charged surfaces was probably first ex ...
. Following the analogy of digital microelectronics, this approach is referred to as digital microfluidics. Le Pesant et al. pioneered the use of electrocapillary forces to move droplets on a digital track. The "fluid transistor" pioneered by Cytonix also played a role. The technology was subsequently commercialised by Duke University. By using discrete unit-volume droplets, a microfluidic function can be reduced to a set of repeated basic operations, i.e., moving one unit of fluid over one unit of distance. This "digitisation" method facilitates the use of a hierarchical and cell-based approach for microfluidic biochip design. Therefore, digital microfluidics offers a flexible and scalable system architecture as well as high
fault-tolerance Fault tolerance is the property that enables a system to continue operating properly in the event of the failure of one or more faults within some of its components. If its operating quality decreases at all, the decrease is proportional to the ...
capability. Moreover, because each droplet can be controlled independently, these systems also have dynamic reconfigurability, whereby groups of unit cells in a microfluidic array can be reconfigured to change their functionality during the concurrent execution of a set of bioassays. Although droplets are manipulated in confined microfluidic channels, since the control on droplets is not independent, it should not be confused as "digital microfluidics". One common actuation method for digital microfluidics is
electrowetting Electrowetting is the modification of the wetting properties of a surface (which is typically hydrophobic) with an applied electric field. History The electrowetting of mercury and other liquids on variably charged surfaces was probably first ex ...
-on-dielectric ( EWOD). Many lab-on-a-chip applications have been demonstrated within the digital microfluidics paradigm using electrowetting. However, recently other techniques for droplet manipulation have also been demonstrated using magnetic force,
surface acoustic wave A surface acoustic wave (SAW) is an acoustic wave traveling along the surface of a material exhibiting elasticity, with an amplitude that typically decays exponentially with depth into the material, such that they are confined to a depth of about ...
s,
optoelectrowetting Optoelectrowetting (OEW) is a method of liquid droplet manipulation used in microfluidics applications. This technique builds on the principle of electrowetting, which has proven useful in liquid actuation due to fast switching response times and lo ...
, mechanical actuation, etc.


Paper-based microfluidics

Paper-based microfluidic devices fill a growing niche for portable, cheap, and user-friendly medical diagnostic systems. Paper based microfluidics rely on the phenomenon of capillary penetration in porous media. To tune fluid penetration in porous substrates such as paper in two and three dimensions, the pore structure, wettability and geometry of the microfluidic devices can be controlled while the viscosity and evaporation rate of the liquid play a further significant role. Many such devices feature hydrophobic barriers on hydrophilic paper that passively transport aqueous solutions to outlets where biological reactions take place. Paper-based microfluidics are considered as portable point-of-care biosensors used in a remote setting where advanced medical diagnostic tools are not accessible. Current applications include portable glucose detection and environmental testing, with hopes of reaching areas that lack advanced medical diagnostic tools.


Particle detection microfluidics

One application area that has seen significant academic effort and some commercial effort is in the area of particle detection in fluids. Particle detection of small fluid-borne particles down to about 1 μm in diameter is typically done using a
Coulter counter A Coulter counter is an apparatus for counting and sizing particles suspended in electrolytes. The Coulter counter is the commercial term for the technique known as resistive pulse sensing or electrical zone sensing, the apparatus is based on ...
, in which electrical signals are generated when a weakly-conducting fluid such as in saline water is passed through a small (~100 μm diameter) pore, so that an electrical signal is generated that is directly proportional to the ratio of the particle volume to the pore volume. The physics behind this is relatively simple, described in a classic paper by DeBlois and Bean, and the implementation first described in Coulter's original patent. This is the method used to e.g. size and count erythrocytes (red blood cells
iki IKI may refer to: * Internationales Kulturinstitut in Vienna * Iodine potassium-iodide, a chemical compound * Russian Space Research Institute originally known as IKI RAN * Iki Airport, IATA code Iki or iki may refer to: * Iki Island, a Japanese ...
as well as leukocytes ( white blood cells) for standard blood analysis. The generic term for this method is
resistive pulse sensing Resistive pulse sensing (RPS) is the generic, non-commercial term given for the well-developed technology used to detect, and measure the size of, individual particles in fluid. First invented by Wallace H. Coulter in 1953, the RPS technique is the ...
(RPS); Coulter counting is a trademark term. However, the RPS method does not work well for particles below 1 μm diameter, as the signal-to-noise ratio falls below the reliably detectable limit, set mostly by the size of the pore in which the analyte passes and the input noise of the first-stage
amplifier An amplifier, electronic amplifier or (informally) amp is an electronic device that can increase the magnitude of a signal (a time-varying voltage or current). It may increase the power significantly, or its main effect may be to boost t ...
. The limit on the pore size in traditional RPS Coulter counters is set by the method used to make the pores, which while a trade secret, most likely uses traditional mechanical methods. This is where microfluidics can have an impact: The
lithography Lithography () is a planographic method of printing originally based on the immiscibility of oil and water. The printing is from a stone (lithographic limestone) or a metal plate with a smooth surface. It was invented in 1796 by the German a ...
-based production of microfluidic devices, or more likely the production of reusable molds for making microfluidic devices using a molding process, is limited to sizes much smaller than traditional machining. Critical dimensions down to 1 μm are easily fabricated, and with a bit more effort and expense, feature sizes below 100 nm can be patterned reliably as well. This enables the inexpensive production of pores integrated in a microfluidic circuit where the pore diameters can reach sizes of order 100 nm, with a concomitant reduction in the minimum particle diameters by several orders of magnitude. As a result there has been some university-based development of microfluidic particle counting and sizing with the accompanying commercialization of this technology. This method has been termed microfluidic
resistive pulse sensing Resistive pulse sensing (RPS) is the generic, non-commercial term given for the well-developed technology used to detect, and measure the size of, individual particles in fluid. First invented by Wallace H. Coulter in 1953, the RPS technique is the ...
(MRPS).


Microfluidic-assisted magnetophoresis

One major area of application for microfluidic devices is the separation and sorting of different fluids or cell types. Recent developments in the microfluidics field have seen the integration of microfluidic devices with magnetophoresis: the migration of particles by a magnetic field. This can be accomplished by sending a fluid containing at least one magnetic component through a microfluidic channel that has a
magnet A magnet is a material or object that produces a magnetic field. This magnetic field is invisible but is responsible for the most notable property of a magnet: a force that pulls on other ferromagnetic materials, such as iron, steel, nicke ...
positioned along the length of the channel. This creates a magnetic field inside the microfluidic channel which draws
magnetically Magnetism is the class of physical attributes that are mediated by a magnetic field, which refers to the capacity to induce attractive and repulsive phenomena in other entities. Electric currents and the magnetic moments of elementary particles ...
active substances towards it, effectively separating the magnetic and non-magnetic components of the fluid. This technique can be readily utilized in
industrial Industrial may refer to: Industry * Industrial archaeology, the study of the history of the industry * Industrial engineering, engineering dealing with the optimization of complex industrial processes or systems * Industrial city, a city dominate ...
settings where the fluid at hand already contains magnetically active material. For example, a handful of metallic impurities can find their way into certain consumable liquids, namely
milk Milk is a white liquid food produced by the mammary glands of mammals. It is the primary source of nutrition for young mammals (including breastfed human infants) before they are able to digest solid food. Immune factors and immune-modula ...
and other dairy products. Conveniently, in the case of milk, many of these metal contaminants exhibit
paramagnetism Paramagnetism is a form of magnetism whereby some materials are weakly attracted by an externally applied magnetic field, and form internal, induced magnetic fields in the direction of the applied magnetic field. In contrast with this behavior, ...
. Therefore, before packaging, milk can be flowed through channels with magnetic gradients as a means of purifying out the metal contaminants. Other, more research-oriented applications of microfluidic-assisted magnetophoresis are numerous and are generally targeted towards
cell Cell most often refers to: * Cell (biology), the functional basic unit of life Cell may also refer to: Locations * Monastic cell, a small room, hut, or cave in which a religious recluse lives, alternatively the small precursor of a monastery ...
separation. The general way this is accomplished involves several steps. First, a paramagnetic substance (usually micro/
nanoparticle A nanoparticle or ultrafine particle is usually defined as a particle of matter that is between 1 and 100 nanometres (nm) in diameter. The term is sometimes used for larger particles, up to 500 nm, or fibers and tubes that are less than 10 ...
s or a paramagnetic fluid) needs to be functionalized to target the cell type of interest. This can be accomplished by identifying a transmembranal protein unique to the cell type of interest and subsequently functionalizing magnetic particles with the complementary
antigen In immunology, an antigen (Ag) is a molecule or molecular structure or any foreign particulate matter or a pollen grain that can bind to a specific antibody or T-cell receptor. The presence of antigens in the body may trigger an immune respons ...
or
antibody An antibody (Ab), also known as an immunoglobulin (Ig), is a large, Y-shaped protein used by the immune system to identify and neutralize foreign objects such as pathogenic bacteria and viruses. The antibody recognizes a unique molecule of the ...
. Once the magnetic particles are functionalized, they are dispersed in a cell mixture where they bind to only the cells of interest. The resulting cell/particle mixture can then be flowed through a microfluidic device with a magnetic field to separate the targeted cells from the rest. Conversely, microfluidic-assisted magnetophoresis may be used to facilitate efficient mixing within microdroplets or plugs. To accomplish this, microdroplets are injected with paramagnetic nanoparticles and are flowed through a straight channel which passes through rapidly alternating magnetic fields. This causes the magnetic particles to be quickly pushed from side to side within the droplet and results in the mixing of the microdroplet contents. This eliminates the need for tedious engineering considerations that are necessary for traditional, channel-based droplet mixing. Other research has also shown that the label-free separation of cells may be possible by suspending cells in a paramagnetic fluid and taking advantage of the magneto-Archimedes effect. While this does eliminate the complexity of particle functionalization, more research is needed to fully understand the magneto-Archimedes phenomenon and how it can be used to this end. This is not an exhaustive list of the various applications of microfluidic-assisted magnetophoresis; the above examples merely highlight the versatility of this separation technique in both current and future applications.


Key application areas

Microfluidic structures include micropneumatic systems, i.e. microsystems for the handling of off-chip fluids (liquid pumps, gas valves, etc.), and microfluidic structures for the on-chip handling of nanoliter (nl) and picoliter (pl) volumes. To date, the most successful commercial application of microfluidics is the inkjet printhead. Additionally, microfluidic manufacturing advances mean that makers can produce the devices in low-cost plastics and automatically verify part quality. Advances in microfluidics technology are revolutionizing
molecular biology Molecular biology is the branch of biology that seeks to understand the molecular basis of biological activity in and between cells, including biomolecular synthesis, modification, mechanisms, and interactions. The study of chemical and physi ...
procedures for enzymatic analysis (e.g.,
glucose Glucose is a simple sugar with the molecular formula . Glucose is overall the most abundant monosaccharide, a subcategory of carbohydrates. Glucose is mainly made by plants and most algae during photosynthesis from water and carbon dioxide, u ...
and lactate assays), DNA analysis (e.g.,
polymerase chain reaction The polymerase chain reaction (PCR) is a method widely used to rapidly make millions to billions of copies (complete or partial) of a specific DNA sample, allowing scientists to take a very small sample of DNA and amplify it (or a part of it) ...
and high-throughput sequencing), proteomics, and in chemical synthesis. The basic idea of microfluidic biochips is to integrate assay operations such as detection, as well as sample pre-treatment and sample preparation on one chip. An emerging application area for biochips is
clinical pathology Clinical pathology is a medical specialty that is concerned with the diagnosis of disease based on the laboratory analysis of bodily fluids, such as blood, urine, and tissue homogenates or extracts using the tools of chemistry, microbiology, h ...
, especially the immediate
point-of-care Clinical point of care (POC) is the point in time when clinicians deliver healthcare products and services to patients at the time of care. Clinical documentation Clinical documentation is a record of the critical thinking and judgment of a heal ...
diagnosis of diseases. In addition, microfluidics-based devices, capable of continuous sampling and real-time testing of air/water samples for biochemical toxins and other dangerous
pathogens In biology, a pathogen ( el, πάθος, "suffering", "passion" and , "producer of") in the oldest and broadest sense, is any organism or agent that can produce disease. A pathogen may also be referred to as an infectious agent, or simply a ger ...
, can serve as an always-on "bio-smoke alarm" for early warning. Microfluidic technology has led to the creation of powerful tools for biologists to control the complete cellular environment, leading to new questions and discoveries. Many diverse advantages of this technology for microbiology are listed below: * General single cell studies including growth * Cellular aging: microfluidic devices such as the "mother machine" allow tracking of thousands of individual cells for many generations until they die * Microenvironmental control: ranging from mechanical environment to chemical environment * Precise spatiotemporal concentration gradients by incorporating multiple chemical inputs to a single device * Force measurements of adherent cells or confined chromosomes: objects trapped in a microfluidic device can be directly manipulated using optical tweezers or other force-generating methods * Confining cells and exerting controlled forces by coupling with external force-generation methods such as Stokes flow, optical tweezer, or controlled deformation of the PDMS (
Polydimethylsiloxane Polydimethylsiloxane (PDMS), also known as dimethylpolysiloxane or dimethicone, belongs to a group of polymeric organosilicon compounds that are commonly referred to as silicones. PDMS is the most widely used silicon-based organic polymer, as its ...
) device * Electric field integration * Plant on a chip and plant tissue culture * Antibiotic resistance: microfluidic devices can be used as heterogeneous environments for microorganisms. In a heterogeneous environment, it is easier for a microorganism to evolve. This can be useful for testing the acceleration of evolution of a microorganism / for testing the development of antibiotic resistance. Some of these areas are further elaborated in the sections below:


DNA chips (microarrays)

Early biochips were based on the idea of a DNA microarray, e.g., the GeneChip DNAarray from Affymetrix, which is a piece of glass, plastic or silicon substrate, on which pieces of DNA (probes) are affixed in a microscopic array. Similar to a DNA microarray, a protein array is a miniature array where a multitude of different capture agents, most frequently monoclonal antibodies, are deposited on a chip surface; they are used to determine the presence and/or amount of
protein Proteins are large biomolecules and macromolecules that comprise one or more long chains of amino acid residues. Proteins perform a vast array of functions within organisms, including catalysing metabolic reactions, DNA replication, res ...
s in biological samples, e.g.,
blood Blood is a body fluid in the circulatory system of humans and other vertebrates that delivers necessary substances such as nutrients and oxygen to the cells, and transports metabolic waste products away from those same cells. Blood in the cir ...
. A drawback of DNA and protein arrays is that they are neither reconfigurable nor scalable after manufacture. Digital microfluidics has been described as a means for carrying out
Digital PCR Digital polymerase chain reaction (digital PCR, DigitalPCR, dPCR, or dePCR) is a biotechnological refinement of conventional polymerase chain reaction methods that can be used to directly quantify and clonally amplify nucleic acids strands includ ...
.


Molecular biology

In addition to microarrays, biochips have been designed for two-dimensional electrophoresis,
transcriptome The transcriptome is the set of all RNA transcripts, including coding and non-coding, in an individual or a population of cells. The term can also sometimes be used to refer to all RNAs, or just mRNA, depending on the particular experiment. The t ...
analysis, and PCR amplification. Other applications include various electrophoresis and
liquid chromatography In chemical analysis, chromatography is a laboratory technique for the separation of a mixture into its components. The mixture is dissolved in a fluid solvent (gas or liquid) called the ''mobile phase'', which carries it through a system (a ...
applications for proteins and DNA, cell separation, in particular, blood cell separation, protein analysis, cell manipulation and analysis including cell viability analysis and
microorganism A microorganism, or microbe,, ''mikros'', "small") and ''organism'' from the el, ὀργανισμός, ''organismós'', "organism"). It is usually written as a single word but is sometimes hyphenated (''micro-organism''), especially in olde ...
capturing.


Evolutionary biology

By combining microfluidics with
landscape ecology Landscape ecology is the science of studying and improving relationships between ecological processes in the environment and particular ecosystems. This is done within a variety of landscape scales, development spatial patterns, and organizatio ...
and
nanofluidics Nanofluidics is the study of the behavior, manipulation, and control of fluids that are confined to structures of nanometer (typically 1–100 nm) characteristic dimensions (1 nm = 10−9 m). Fluids confined in these structures exhibit p ...
, a nano/micro fabricated fluidic landscape can be constructed by building local patches of
bacterial Bacteria (; singular: bacterium) are ubiquitous, mostly free-living organisms often consisting of one biological cell. They constitute a large domain of prokaryotic microorganisms. Typically a few micrometres in length, bacteria were amon ...
habitat In ecology, the term habitat summarises the array of resources, physical and biotic factors that are present in an area, such as to support the survival and reproduction of a particular species. A species habitat can be seen as the physical ...
and connecting them by dispersal corridors. The resulting landscapes can be used as physical implementations of an
adaptive landscape Adaptation, in biology, is the process or trait by which organisms or population better match their environment Adaptation may also refer to: Arts * Adaptation (arts), a transfer of a work of art from one medium to another ** Film adaptation, a ...
, by generating a spatial mosaic of patches of opportunity distributed in space and time. The patchy nature of these fluidic landscapes allows for the study of adapting bacterial cells in a metapopulation system. The evolutionary ecology of these bacterial systems in these synthetic ecosystems allows for using
biophysics Biophysics is an interdisciplinary science that applies approaches and methods traditionally used in physics to study biological phenomena. Biophysics covers all scales of biological organization, from molecular to organismic and populations. ...
to address questions in
evolutionary biology Evolutionary biology is the subfield of biology that studies the evolutionary processes ( natural selection, common descent, speciation) that produced the diversity of life on Earth. It is also defined as the study of the history of life ...
.


Cell behavior

The ability to create precise and carefully controlled chemoattractant gradients makes microfluidics the ideal tool to study motility, chemotaxis and the ability to evolve / develop resistance to antibiotics in small populations of microorganisms and in a short period of time. These microorganisms including
bacteria Bacteria (; singular: bacterium) are ubiquitous, mostly free-living organisms often consisting of one Cell (biology), biological cell. They constitute a large domain (biology), domain of prokaryotic microorganisms. Typically a few micrometr ...
and the broad range of organisms that form the marine
microbial loop The microbial loop describes a trophic pathway where, in aquatic systems, dissolved organic carbon (DOC) is returned to higher trophic levels via its incorporation into bacterial biomass, and then coupled with the classic food chain formed by phy ...
, responsible for regulating much of the oceans' biogeochemistry. Microfluidics has also greatly aided the study of
durotaxis Durotaxis is a form of cell migration in which cells are guided by rigidity gradients, which arise from differential structural properties of the extracellular matrix (ECM). Most normal cells migrate up rigidity gradients (in the direction of great ...
by facilitating the creation of durotactic (stiffness) gradients.


Cellular biophysics

By rectifying the motion of individual swimming bacteria, microfluidic structures can be used to extract mechanical motion from a population of motile bacterial cells. This way, bacteria-powered rotors can be built.


Optics

The merger of microfluidics and optics is typical known as optofluidics. Examples of optofluidic devices are tunable microlens arrays and optofluidic microscopes. Microfluidic flow enables fast sample throughput, automated imaging of large sample populations, as well as 3D capabilities. or superresolution.


High Performance Liquid Chromatography (HPLC)

HPLC in the field of microfluidics comes in two different forms. Early designs included running liquid through the HPLC column then transferring the eluted liquid to microfluidic chips and attaching HPLC columns to the microfluidic chip directly. The early methods had the advantage of easier detection from certain machines like those that measure fluorescence. More recent designs have fully integrated HPLC columns into microfluidic chips. The main advantage of integrating HPLC columns into microfluidic devices is the smaller form factor that can be achieved, which allows for additional features to be combined within one microfluidic chip. Integrated chips can also be fabricated from multiple different materials, including glass and polyimide which are quite different from the standard material of PDMS used in many different droplet-based microfluidic devices. This is an important feature because different applications of HPLC microfluidic chips may call for different pressures. PDMS fails in comparison for high-pressure uses compared to glass and polyimide. High versatility of HPLC integration ensures robustness by avoiding connections and fittings between the column and chip. The ability to build off said designs in the future allows the field of microfluidics to continue expanding its potential applications. The potential applications surrounding integrated HPLC columns within microfluidic devices have proven expansive over the last 10–15 years. The integration of such columns allows for experiments to be run where materials were in low availability or very expensive, like in biological analysis of proteins. This reduction in reagent volumes allows for new experiments like single-cell protein analysis, which due to size limitations of prior devices, previously came with great difficulty. The coupling of HPLC-chip devices with other spectrometry methods like mass-spectrometry allow for enhanced confidence in identification of desired species, like proteins. Microfluidic chips have also been created with internal delay-lines that allow for gradient generation to further improve HPLC, which can reduce the need for further separations. Some other practical applications of integrated HPLC chips include the determination of drug presence in a person through their hair and the labeling of peptides through reverse phase liquid chromatography.


Acoustic droplet ejection (ADE)

Acoustic droplet ejection uses a pulse of
ultrasound Ultrasound is sound waves with frequencies higher than the upper audible limit of human hearing. Ultrasound is not different from "normal" (audible) sound in its physical properties, except that humans cannot hear it. This limit varies ...
to move low volumes of
fluids In physics, a fluid is a liquid, gas, or other material that continuously deforms (''flows'') under an applied shear stress, or external force. They have zero shear modulus, or, in simpler terms, are substances which cannot resist any she ...
(typically nanoliters or picoliters) without any physical contact. This technology focuses acoustic energy into a fluid sample to eject droplets as small as a millionth of a millionth of a litre (picoliter = 10−12 litre). ADE technology is a very gentle process, and it can be used to transfer proteins, high molecular weight DNA and live cells without damage or loss of viability. This feature makes the technology suitable for a wide variety of applications including proteomics and cell-based assays.


Fuel cells

Microfluidic fuel cells can use laminar flow to separate the fuel and its oxidant to control the interaction of the two fluids without the physical barrier that conventional fuel cells require.


Astrobiology

To understand the prospects for life to exist elsewhere in the universe, astrobiologists are interested in measuring the chemical composition of extraplanetary bodies. Because of their small size and wide-ranging functionality, microfluidic devices are uniquely suited for these remote sample analyses. From an extraterrestrial sample, the organic content can be assessed using microchip
capillary electrophoresis Capillary electrophoresis (CE) is a family of electrokinetic separation methods performed in submillimeter diameter capillaries and in micro- and nanofluidic channels. Very often, CE refers to capillary zone electrophoresis (CZE), but other elect ...
and selective fluorescent dyes. These devices are capable of detecting
amino acid Amino acids are organic compounds that contain both amino and carboxylic acid functional groups. Although hundreds of amino acids exist in nature, by far the most important are the alpha-amino acids, which comprise proteins. Only 22 alpha a ...
s,
peptide Peptides (, ) are short chains of amino acids linked by peptide bonds. Long chains of amino acids are called proteins. Chains of fewer than twenty amino acids are called oligopeptides, and include dipeptides, tripeptides, and tetrapeptides. ...
s,
fatty acid In chemistry, particularly in biochemistry, a fatty acid is a carboxylic acid with an aliphatic chain, which is either saturated or unsaturated. Most naturally occurring fatty acids have an unbranched chain of an even number of carbon atoms, ...
s, and simple
aldehyde In organic chemistry, an aldehyde () is an organic compound containing a functional group with the structure . The functional group itself (without the "R" side chain) can be referred to as an aldehyde but can also be classified as a formyl grou ...
s, ketones, and
thiol In organic chemistry, a thiol (; ), or thiol derivative, is any organosulfur compound of the form , where R represents an alkyl or other organic substituent. The functional group itself is referred to as either a thiol group or a sulfhydryl gro ...
s. These analyses coupled together could allow powerful detection of the key components of life, and hopefully inform our search for functioning extraterrestrial life.


Food Science

Microfluidic techniques such as droplet microfluidics, paper microfluidics, and
lab-on-a-chip A lab-on-a-chip (LOC) is a device that integrates one or several laboratory functions on a single integrated circuit (commonly called a "chip") of only millimeters to a few square centimeters to achieve automation and high-throughput screening. ...
are used in the realm of food science in a variety of categories. Research in nutrition, food processing, and food safety benefit from microfluidic technique because experiments can be done with less reagents. Food processing requires the ability to enable shelf stability in foods, such as emulsions or additions of preservatives. Techniques such as droplet microfluidics are used to create emulsions that are more controlled and complex than those created by traditional homogenization due to the precision of droplets that is achievable. Using microfluidics for emulsions is also more energy efficient compared to homogenization in which “only 5% of the supplied energy is used to generate the emulsion, with the rest dissipated as heat” . Although these methods have benefits, they currently lack the ability to be produced at large scale that is needed for commercialization. Microfluidics are also used in research as they allow for innovation in food chemistry and food processing. An example in food engineering research is a novel micro-3D-printed device fabricated to research production of droplets for potential food processing industry use, particularly in work with enhancing emulsions. Paper and droplet microfluidics allow for devices that can detect small amounts of unwanted bacteria or chemicals, making them useful in food safety and analysis.Harmon JB, Gray HK, Young CC, Schwab KJ (2020) Microfluidic droplet application for bacterial surveillance in fresh-cut produce wash waters. PLoS ONE 15(6): e0233239. https://doi.org/10.1371/journal.pone.0233239 Paper-based microfluidic devices are often referred to as microfluidic paper-based analytical devices (µPADs) and can detect such things as nitrate, preservatives, or antibiotics in meat by a colorimetric reaction that can be detected with a smartphone. These methods are being researched because they use less reactants, space, and time compared to traditional techniques such as liquid chromatography. µPADs also make home detection tests possible , which is of interest to those with allergies and intolerances. In addition to paper-based methods, research demonstrates droplet-based microfluidics shows promise in drastically shortening the time necessary to confirm viable bacterial contamination in agricultural waters in the domestic and international food industry.


Future directions


Microfluidics for personalized cancer treatment

Personalized cancer treatment is an tuned method based on the patient's diagnosis and background. Microfluidic technology offers sensitive detection with higher throughput, as well as reduced time and costs. For personalized cancer treatment, tumor composition and drug sensitivities are very important. A patient's drug response can be predicted based on the status of
biomarker In biomedical contexts, a biomarker, or biological marker, is a measurable indicator of some biological state or condition. Biomarkers are often measured and evaluated using blood, urine, or soft tissues to examine normal biological processes, p ...
s, or the severity and progression of the disease can be predicted based on the atypical presence of specific cells. Drop-
qPCR A real-time polymerase chain reaction (real-time PCR, or qPCR) is a laboratory technique of molecular biology based on the polymerase chain reaction (PCR). It monitors the amplification of a targeted DNA molecule during the PCR (i.e., in real ...
is a droplet microfluidic technology in which droplets are transported in a reusable capillary and alternately flow through two areas maintained at different constant temperatures and fluorescence detection. It can be efficient with a low contamination risk to detect
Her2 Receptor tyrosine-protein kinase erbB-2 is a protein that in humans is encoded by the ''ERBB2'' gene. ERBB is abbreviated from erythroblastic oncogene B, a gene originally isolated from the avian genome. The human protein is also frequently refer ...
. A digital droplet‐based PCR method can be used to detect the
KRAS ''KRAS'' ( Kirsten rat sarcoma virus) is a gene that provides instructions for making a protein called K-Ras, a part of the RAS/MAPK pathway. The protein relays signals from outside the cell to the cell's nucleus. These signals instruct the cel ...
mutations with TaqMan probes, to enhance detection of the mutative gene ratio. In addition, accurate prediction of postoperative disease progression in
breast The breast is one of two prominences located on the upper ventral region of a primate's torso. Both females and males develop breasts from the same embryological tissues. In females, it serves as the mammary gland, which produces and sec ...
 or prostate cancer patients is essential for determining post-surgery treatment. A simple microfluidic chamber, coated with a carefully formulated extracellular matrix mixture. is used for cells obtained from tumor
biopsy A biopsy is a medical test commonly performed by a surgeon, interventional radiologist, or an interventional cardiologist. The process involves extraction of sample cells or tissues for examination to determine the presence or extent of a dise ...
after 72 hours of growth and a thorough evaluation of cells by imaging. Microfluidics is also suitable for circulating tumor cells (CTCs) and non- CTCs
liquid biopsy A liquid biopsy, also known as fluid biopsy or fluid phase biopsy, is the sampling and analysis of non-solid biological tissue, primarily blood. Like traditional biopsy, this type of technique is mainly used as a diagnostic and monitoring tool for ...
analysis. Beads conjugate to anti‐ epithelial cell adhesion molecule (EpCAM) antibodies for positive selection in the CTCs isolation chip (iCHIP). CTCs can also be detected by using the acidification of the
tumor microenvironment The tumor microenvironment (TME) is the environment around a tumor, including the surrounding blood vessels, immune cells, fibroblasts, signaling molecules and the extracellular matrix (ECM). The tumor and the surrounding microenvironment are ...
and the difference in membrane capacitance. CTCs are isolated from blood by a microfluidic device, and are cultured on-chip, which can be a method to capture more biological information in a single analysis. For example, it can be used to test the cell survival rate of 40 different drugs or drug combinations. Tumor‐derived
extracellular vesicle Extracellular vesicles (EVs) are lipid bilayer-delimited particles that are naturally released from almost all types of cell but, unlike a cell, cannot replicate. EVs range in diameter from near the size of the smallest physically possible unil ...
s can be isolated from urine and detected by an integrated double‐filtration microfluidic device; they also can be isolated from blood and detected by electrochemical sensing method with a two‐level amplification enzymatic assay. Tumor materials can directly be used for detection through microfluidic devices. To screen
primary cell A primary battery or primary cell is a battery (a galvanic cell) that is designed to be used once and discarded, and not recharged with electricity and reused like a secondary cell (rechargeable battery). In general, the electrochemical reaction ...
s for drugs, it is often necessary to distinguish cancerous cells from non-cancerous cells. A microfluidic chip based on the capacity of cells to pass small constrictions can sort the cell types,
metastases Metastasis is a pathogenic agent's spread from an initial or primary site to a different or secondary site within the host's body; the term is typically used when referring to metastasis by a cancerous tumor. The newly pathological sites, then ...
. Droplet‐based microfluidic devices have the potential to screen different drugs or combinations of drugs, directly on the
primary tumor A primary tumor is a tumor growing at the anatomical site where tumor progression began and proceeded to yield a cancerous mass. Most cancers develop at their primary site but then go on to metastasize or spread to other parts of the body. These fur ...
sample with high accuracy. To improve this strategy, the microfluidic program with a sequential manner of drug cocktails, coupled with fluorescent barcodes, is more efficient. Another advanced strategy is detecting growth rates of single-cell by using suspended microchannel resonators, which can predict drug sensitivities of rare CTCs. Microfluidics devices also can simulate the
tumor microenvironment The tumor microenvironment (TME) is the environment around a tumor, including the surrounding blood vessels, immune cells, fibroblasts, signaling molecules and the extracellular matrix (ECM). The tumor and the surrounding microenvironment are ...
, to help to test anticancer drugs. Microfluidic devices with 2D or
3D cell culture 3-D, 3D, or 3d may refer to: Science, technology, and mathematics Relating to three-dimensionality * Three-dimensional space ** 3D computer graphics, computer graphics that use a three-dimensional representation of geometric data ** 3D film, a ...
s can be used to analyze spheroids for different cancer systems (such as
lung cancer Lung cancer, also known as lung carcinoma (since about 98–99% of all lung cancers are carcinomas), is a malignant lung tumor characterized by uncontrolled cell growth in tissues of the lung. Lung carcinomas derive from transformed, malign ...
and
ovarian cancer Ovarian cancer is a cancerous tumor of an ovary. It may originate from the ovary itself or more commonly from communicating nearby structures such as fallopian tubes or the inner lining of the abdomen. The ovary is made up of three different c ...
), and are essential for multiple anti-cancer drugs and toxicity tests. This strategy can be improved by increasing the throughput and production of spheroids. For example, one droplet-based microfluidic device for
3D cell culture 3-D, 3D, or 3d may refer to: Science, technology, and mathematics Relating to three-dimensionality * Three-dimensional space ** 3D computer graphics, computer graphics that use a three-dimensional representation of geometric data ** 3D film, a ...
produces 500 spheroids per chip. These spheroids can be cultured longer in different surroundings to analyze and monitor. The other advanced technology is organs‐on‐a‐chip, and it can be used to simulate several organs to determine the drug metabolism and activity based on vessels mimicking, as well as mimic pH,
oxygen Oxygen is the chemical element with the symbol O and atomic number 8. It is a member of the chalcogen group in the periodic table, a highly reactive nonmetal, and an oxidizing agent that readily forms oxides with most elements as ...
... to analyze the relationship between drugs and human organ surroundings. A recent strategy is single-cell chromatin immunoprecipitation (ChiP)‐Sequencing in
droplets A drop or droplet is a small column of liquid, bounded completely or almost completely by free surfaces. A drop may form when liquid accumulates at the lower end of a tube or other surface boundary, producing a hanging drop called a pendant d ...
, which operates by combining droplet‐based single cell
RNA sequencing RNA-Seq (named as an abbreviation of RNA sequencing) is a sequencing technique which uses next-generation sequencing (NGS) to reveal the presence and quantity of RNA in a biological sample at a given moment, analyzing the continuously changing ...
with DNA‐barcoded antibodies, possibly to explore the tumor heterogeneity by the genotype and
phenotype In genetics, the phenotype () is the set of observable characteristics or traits of an organism. The term covers the organism's morphology or physical form and structure, its developmental processes, its biochemical and physiological pr ...
to select the personalized anti-cancer drugs and prevent the cancer relapse. Overall, microfluidic techniques have a strong potential for personalized cancer treatment and bringing in new methods for future cancer therapies. However, challenges remain to integrate these methods with clinical treatment, based on the amount of genetic or
biomarker In biomedical contexts, a biomarker, or biological marker, is a measurable indicator of some biological state or condition. Biomarkers are often measured and evaluated using blood, urine, or soft tissues to examine normal biological processes, p ...
's information, unreliable equipment, or difficulty of interfacing with clinical operations.


Microfluidic drug assays:


On-chip characterization:


Microfluidics in the classroom: On-chip acid-base titrations


Sepsis Sepsis, formerly known as septicemia (septicaemia in British English) or blood poisoning, is a life-threatening condition that arises when the body's response to infection causes injury to its own tissues and organs. This initial stage is follo ...
detection in minutes not days.


Unlocking multi-angle imaging for microfluidic devices


See also

*
Advanced Simulation Library Advanced Simulation Library (ASL) is free and open-source hardware-accelerated multiphysics simulation platform. It enables users to write customized numerical solvers in C++ and deploy them on a variety of massively parallel architecture ...
* Droplet-based microfluidics *
Fluidics Fluidics, or fluidic logic, is the use of a fluid to perform analog or digital operations similar to those performed with electronics. The physical basis of fluidics is pneumatics and hydraulics, based on the theoretical foundation of fluid dyna ...
*
Microphysiometry Microphysiometry is the ''in vitro'' measurement of the functions and activities of life or of living matter (as organs, tissues, or cells) and of the physical and chemical phenomena involved on a very small (micrometer) scale. The term microphysiom ...
*
Micropump Micropumps are devices that can control and manipulate small fluid volumes. Although any kind of small pump is often referred to as micropump, a more accurate definition restricts this term to pumps with functional dimensions in the micrometer ran ...
s * Microvalves * Induced-charge electrokinetics *
Lab-on-a-chip A lab-on-a-chip (LOC) is a device that integrates one or several laboratory functions on a single integrated circuit (commonly called a "chip") of only millimeters to a few square centimeters to achieve automation and high-throughput screening. ...
* uFluids@Home *
Paper-based microfluidics Paper-based microfluidics are microfluidic devices that consist of a series of hydrophilic cellulose or nitrocellulose fibers that transport fluid from an inlet through the porous medium to a desired outlet or region of the device, by means of capi ...
* Microfluidic cell culture * Microfluidic Modulation Spectroscopy


References


Further reading


Review papers

* * * * * * * * * *


Books

* * Folch, Albert. ''Hidden in Plain Sight: The History, Science, and Engineering of Microfluidic Technology'' (MIT Press, 2022
online review
* * * * *


Education

{{Genomics Microfluidics Fluid dynamics Nanotechnology Biotechnology Gas technologies