Accurate and high throughput cell sorting is a critical enabling technology in molecular and cellular biology, biotechnology, and medicine While conventional methods can provide high efficiency sorting in short timescales, advances in microfluidics have enabled the realization of miniaturized devices offering similar capabilities that exploit a variety of physical principles We classify these technologies as either active or passive Active systems generally use external fields (eg, acoustic, electric, magnetic, and optical) to impose forces to displace cells for sorting, whereas passive systems use inertial forces, filters, and adhesion mechanisms to purify cell populations Cell sorting on microchips provides numerous advantages over conventional methods by reducing the size of necessary equipment, eliminating potentially biohazardous aerosols, and simplifying the complex protocols commonly associated with cell sorting Additionally, microchip devices are well suited for parallelization, enabling complete lab-on-a-chip devices for cellular isolation, analysis, and experimental processing In this review, we examine the breadth of microfluidic cell sorting technologies, while focusing on those that offer the greatest potential for translation into clinical and industrial practice and that offer multiple, useful functions We organize these sorting technologies by the type of cell preparation required (ie, fluorescent label-based sorting, bead-based sorting, and label-free sorting) as well as by the physical principles underlying each sorting mechanism

Microfluidic cell sorting: a review of the advances in the separation of cells from debulking to rare cell isolation

These guidelines are a consensus work of a considerable number of members of the immunology and flow cytometry community. They provide the theory and key practical aspects of flow cytometry enabling immunologists to avoid the common errors that often undermine immunological data. Notably, there are comprehensive sections of all major immune cell types with helpful Tables detailing phenotypes in murine and human cells. The latest flow cytometry techniques and applications are also described, featuring examples of the data that can be generated and, importantly, how the data can be analysed. Furthermore, there are sections detailing tips, tricks and pitfalls to avoid, all written and peer-reviewed by leading experts in the field, making this an essential research companion.

Guidelines for the use of flow cytometry and cell sorting in immunological studies (second edition)

The marriage between immunology and cytometry is one of the most stable and productive in the recent history of science. A rapid search in PubMed shows that, as of July 2017, using “flow cytometry immunology” as a search term yields more than 68 000 articles, the first of which, interestingly, is not about lymphocytes. It might be stated that, after a short engagement, the exchange of the wedding rings between immunology and cytometry officially occurred when the idea to link fluorochromes to monoclonal antibodies came about. After this, recognizing different types of cells became relatively easy and feasible not only by using a simple fluorescence microscope, but also by a complex and sometimes esoteric instrument, the flow cytometer that is able to count hundreds of cells in a single second, and can provide repetitive results in a tireless manner. Given this, the possibility to analyse immune phenotypes in a variety of clinical conditions has changed the use of the flow cytometer, which was incidentally invented in the late 1960s to measure cellular DNA by using intercalating dyes, such as ethidium bromide. The epidemics of HIV/AIDS in the 1980s then gave a dramatic impulse to the technology of counting specific cells, since it became clear that the quantification of the number of peripheral blood CD4+ T cells was crucial to follow the course of the infection, and eventually for monitoring the therapy. As a consequence, the development of flow cytometers that had to be easy-to-use in all clinical laboratories helped to widely disseminate this technology. Nowadays, it is rare to find an immunological paper or read a conference abstract in which the authors did not use flow cytometry as the main tool to dissect the immune system and identify its fine and complex functions. Of note, recent developments have created the sophisticated technology of mass cytometry, which is able to simultaneously identify dozens of molecules at the single cell level and allows us to better understand the complexity and beauty of the immune system.

Guidelines for the use of flow cytometry and cell sorting in immunological studies

Flow cytometry is a complete technology given to biologists to study cellular populations with high precision. This technology elegantly combines sample dimension, data acquisition speed, precision and measurement multiplicity. Beyond the statistical aspect, flow cytometry offers the possibility to physically separate sub-populations. These performances come from the common endeavor of physicists, biophysicists, biologists and computer engineers, who succeeded, by providing new concepts, to bring flow cytometry to current maturity. The aim of this paper is to present a complete retrospective of the technique and remind flow cytometry fundamentals before focusing on recent commercial instrumentation.

/pdf/flow-cytometry-retrospective-fundamentals-and-recent-21rnqesacr.pdf

Flow cytometry: retrospective, fundamentals and recent instrumentation.

Background: Acoustophoresis has been utilized successfully in applications including cell trapping, focusing, and purification. One current limitation of acoustophoresis for cell sorting is the reliance on the inherent physical properties of cells (e.g., compressibility, density) instead of selecting cells based upon biologically relevant surfacepresenting antigens. Introducing an acoustophoretic cell sorting approach that allows biochemical specificity may overcome this limitation, thus advancing the value of acoustophoresis approaches for both the basic research and clinical fields. Results: The results presented herein demonstrate the ability for negative acoustic contrast particles (NACPs) to specifically capture and transport positive acoustic contrast particles (PACPs) to the antinode of an ultrasound standing wave. Emulsification and post curing of pre-polymers, either polydimethylsiloxane (PDMS) or polyvinylmethylsiloxane (PVMS), within aqueous surfactant solution results in the formation of stable NACPs that focus onto pressure antinodes. We used either photochemical reactions with biotin-tetrafluorophenyl azide (biotin-TFPA) or end-functionalization of Pluronic F108 surfactant to biofunctionalize NACPs. These biotinylated NACPs bind specifically to streptavidin polystyrene microparticles (as cell surrogates) and transport them to the pressure antinode within an acoustofluidic chip. Conclusion: To the best of our knowledge, this is the first demonstration of using NACPs as carriers for transport of PACPs in an ultrasound standing wave. By using different silicones (i.e., PDMS, PVMS) and curing chemistries, we demonstrate versatility of silicone materials for NACPs and advance the understanding of useful approaches for preparing NACPs. This bioseparation scheme holds potential for applications requiring rapid, continuous separations such as sorting and analysis of cells and biomolecules.

/pdf/elastomeric-microparticles-for-acoustic-mediated-adcxom6q1f.pdf

Elastomeric microparticles for acoustic mediated bioseparations

Coupling miniature liquid fueled fuel cells with portable electrical devices, with or without rechargeable batteries, lead to new appliance configurations: charging holster, piggyback charger, fuel cell integral with rechargeable battery, fuel cell with voltage regulating electronics, supplying fuel through a tube with a valve or pump, and supplying power though an electric cable between the fuel cell and the electronic device and the fuel cell power pack having independent communication and functions from the portable electrical devices. In all of these configurations there may be a window showing the fuel level of a disposable fuel ampoule or refillable fuel tank. Fuel is distributed through disposable fuel ampoules in blister packages, or refueling bottles. Adjustable moisture and thermal internal insulation allows fuel cells to run at elevated temperatures. These design features permit greater performance, workability, and convenience to the user of these portable electronics. Additionally, these MICRO-FUEL CELL arrays, which pack more energy into a small space, may be mass produced on a plastic film in a reel-to-reel process.

Micro-fuel cell power devices

A hydrogen source system delivers a controlled fuel stream (12) to applications. Said system uses wicking material (10) to control the contact between a mixture of fuel (7) comprising NaBH4, NaOH and H20 contained in a fuel tank (11) and a hydrolysing catalyst (5), to create a feedback mechanism to automatically maintain a constant pressure supply of hydrogen (12). The wicked fuel (7) is bounded by a porous hydrophilic membrane (9) and wets the outer fuel wetting surface (3). The catalyst (5) is attached to hydrophobic porous membrane or molecular filter membrane (1). Production of hydrogen is initiated by the catalyst (5) making contact with surface (3) wetted by the fuel (7). A small compact device packaged for storage, the system operates in any orientation and is mobile.

Portable chemical hydrogen hydride system

A system of two fuel ampoules that can deliver a reactant by diffusion through one of the ampoule walls to the other, such that when said reactant enters the second ampoule, it reacts with another reactant in said second ampoule, making hydrogen gas as a product. Both ampoules are stored in a fuel impermeable container. These ampoules used with small low power fuel cells which need a steady controlled uniform delivery of vaporous fuel such hydrogen and alcohols. This fueling system provides a simple safe fuel interactive system for small hydrogen fuel cells that prevents inadvertent hydrogen production by any single ampoule being exposed to water or typical consumer environments.

Fuel generator with diffusion ampoules for fuel cells

Acoustic cytometry uses radiation pressure forces instead of or in addition to hydrodynamic focusing to position cells or particles in a flowing stream for analysis. Commercial implementations to date combine both hydrodynamic and acoustic focusing together to enable high precision analysis of a broad dynamic range of volumetric sample input rates up to an order of magnitude higher than is practical with hydrodynamic focus alone. This capability allows great flexibility in reducing assay time or modifying or eliminating concentration requirements or concentration steps in sample preparation protocols. It also provides a practical method for processing sub-microliter volumes using sample dilution. In order to take full advantage of this dynamic range, it is necessary to understand the fundamental benefits and limitations of acoustic focusing as applied to flow cytometry. © 2018 by John Wiley & Sons, Inc.

Fundamentals of Acoustic Cytometry

Artificial stoma formed with multilayered structures that actuate with humidity, temperature, chemical environment or light. These actuators can be incorporated into shoes, apparel, fuel cells, machinery, and buildings to control fluid flow or diffusion to regulate humidity, temperature, chemical environment, or light. These actuators can be used as sensors, modify structure, or appearance for greater function, comfort, or aesthetics.

Laminate actuators and valves

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