Metrics & MoreArticle Recommendations CONSPECTUS: Flow cytometry is a powerful tool with applications in diverse fields such as microbiology, immunology, virology, cancer biology, stem cell biology, and metabolic engineering. It rapidly counts and characterizes large heterogeneous populations of cells in suspension (e.g., blood cells, stem cells, cancer cells, and microorganisms) and dissociated solid tissues (e.g., lymph nodes, spleen, and solid tumors) with typical throughputs of 1,000−100,000 events per second (eps). By measuring cell size, cell granularity, and the expression of cell surface and intracellular molecules, it provides systematic insights into biological processes. Flow cytometers may also include cell sorting capabilities to enable subsequent additional analysis of the sorted sample (e.g., electron microscopy and DNA/RNA sequencing), cloning, and directed evolution. Unfortunately, traditional flow cytometry has several critical limitations as it mainly relies on fluorescent labeling for cellular phenotyping, which is an indirect measure of intracellular molecules and surface antigens. Furthermore, it often requires time-consuming preparation protocols and is incompatible with cell therapy. To overcome these difficulties, a different type of flow cytometry based on direct measurements of intracellular molecules by Raman spectroscopy, or "Raman flow cytometry" for short, has emerged. Raman flow cytometry obtains a chemical fingerprint of the cell in a nondestructive manner, allowing for single-cell metabolic phenotyping. However, its slow signal acquisition due to the weak light−molecule interaction of spontaneous Raman scattering prevents the throughput necessary to interrogate large cell populations in reasonable time frames, resulting in throughputs of about 1 eps. The remedy to this throughput limit lies in coherent Raman scattering methods such as stimulated Raman scattering (SRS) and coherent anti-Stokes Raman scattering (CARS), which offer a significantly enhanced light−sample interaction and hence enable high-throughput Raman flow cytometry, Raman imaging flow cytometry, and even Raman image-activated cell sorting (RIACS). In this Account, we outline recent advances, technical challenges, and emerging opportunities of coherent Raman flow cytometry. First, we review the principles of various types of SRS and CARS and introduce several techniques of coherent Raman flow cytometry such as CARS, multiplex CARS, Fourier-transform CARS, SRS, SRS imaging flow cytometry, and RIACS. Next, we discuss a unique set of applications enabled by coherent Raman flow cytometry, from microbiology and lipid biology to cancer detection and cell therapy. Finally, we describe future opportunities and challenges of coherent Raman flow cytometry including increasing sensitivity and throughput, integration with droplet microfluidics, utilizing machine learning techniques, or achieving in vivo flow cytometry. This Account summarizes the growing field of high-throughput Raman flow cytometry and the bright future it ca...
The rich elemental composition, surface chemistry, and outstanding electrical conductivity of MXenes make them a promising class of two-dimensional (2D) materials for electrochemical energy storage. To translate these properties into high performance devices, it is essential to develop fabrication strategies that allow MXenes to be assembled into electrodes with tunable architectures and investigate the effect of their pore structure on the capacitive performance. Here, we report on the fabrication of MXene aerogels with highly ordered lamellar structures by unidirectional freeze-casting of additive-free Ti3C2T x aqueous suspensions. These structures can be subsequently processed into practical supercapacitor electrode films by pressing or calendering steps. This versatile processing route allows a wide control of film thickness, spacing within lamellae (to give electrolyte accessible sites), and densities (over 2 orders of magnitude) and hence gives control over the final properties. The as-prepared MXene aerogel with a density of 13 mg cm–3 achieves 380 F g–1 capacitance at 2 mV s–1 and 75 F g–1 at 50 mV s–1. The calendering of the MXene aerogel into a porous 60 μm thick film with a density of 434 mg cm–3 leads to a superior rate capability of 309 F g–1 at 50 mV s–1. In addition, the rolled electrodes present an improvement in volumetric capacitance of 104 times as compared to the as-prepared MXene aerogel. Finally, the outstanding cyclability of rolled electrodes strengthens their nomination for supercapacitor applications. In this paper we demonstrate the possibilities in tuning the porosity and the electrochemical properties of aerogels highlighting the importance of evaluating new and hybrid processing methods to develop energy storage applications. The simplicity and versatility of the developed fabrication strategy open opportunities for the utilization of MXene lamellae architectures in a wide range of applications requiring controlled porosity including catalysis, filtration, and water purification.
Spiky/hollow metal nanoparticles have applications across a broad range of fields. However, the current bottom-up methods for producing spiky/hollow metal nanoparticles rely heavily on the use of strongly adsorbing surfactant molecules, which is undesirable because these passivate the product particles’ surfaces. Here we report a high-yield surfactant-free synthesis of spiky hollow Au–Ag nanostars (SHAANs). Each SHAAN is composed of >50 spikes attached to a hollow ca. 150 nm diameter cubic core, which makes SHAANs highly plasmonically and catalytically active. Moreover, the surfaces of SHAANs are chemically exposed, which gives them significantly enhanced functionality compared with their surfactant-capped counterparts, as demonstrated in surface-enhanced Raman spectroscopy (SERS) and catalysis. The chemical accessibility of the pristine SHAANs also allows the use of hydroxyethyl cellulose as a weakly bound stabilizing agent. This produces colloidal SHAANs that remain stable for >1 month while retaining the functionalities of the pristine particles and allows even single-particle SERS to be realized.
Microalga-based biomaterial production has attracted attention as a new source of drugs, foods, and biofuels. For enhancing the production efficiency, it is essential to understand its differences between heterogeneous microalgal subpopulations. However, existing techniques are not adequate to address the need due to the lack of single-cell resolution or the inability to perform large-scale analysis and detect small molecules. Here we demonstrated large-scale single-cell analysis of Euglena gracilis (a unicellular microalgal species that produces paramylon as a potential drug for HIV and colon cancer) with our recently developed high-throughput broadband Raman flow cytometer at a throughput of >1,000 cells/s. Specifically, we characterized the intracellular content of paramylon from single-cell Raman spectra of 10,000 E. gracilis cells cultured under five different conditions and found that paramylon contents in E. gracilis cells cultured in an identical condition is given by a log-normal distribution, which is a good model for describing the number of chemicals in a reaction network. The capability of characterizing distribution functions in a label-free manner is an important basis for isolating specific cell populations for synthetic biology via directed evolution based on the intracellular content of metabolites.
Despite an increasingt rend of serviceu seri nvolvement in psychiatric research, few studies involvingt he recipientso fs ecurec are, or individualsw ithl earning disability, were identified. It wasa rgued that service usersw ithl earning disabilitiesd etainedi nas ecureh ospital setting were an important source of information about the care theyr eceived. It wasp redicted that thisg roup would provideav alid and useful account of theire xperiences and that concerns would be raisedi ns imilar areas to thoset hat haveb een reported in other groups of service users. Thesei ncluded concerns relating to environmental conditions, therapeutica ctivities, qualityo fa vailablei nformation about care,a nd concerns relating to livingwithothers.Seven service usersc ompleted as emi-structured interviewa bout their experiences of the care thatt heyr eceived. Data were analysed usingc ontent analysis in order to deriveaseries of keyt hemes whilst acknowledging the individualityofparticipants'experiences.Themes were identified relating to twoa reas:D etention and Treatment. Findings supported predictions that individuals with learning disabilityc ouldg ive valid views about theirt reatment. Therew as overlapb etween the findings of this research and previous studies consideringv iews of mentalh ealth/forensic and learning disabled service users.
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