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In an effort to replace, reduce and refine animal experimentation, there is an unmet need to advance current in vitro models that offer features with physiological relevance and enhanced predictivity of in vivo toxicological output. Hepatic toxicology is key following chemical, drug and nanomaterials (NMs) exposure, as the liver is vital in metabolic detoxification of chemicals as well as being a major site of xenobiotic accumulation (i.e., low solubility particulates). With the ever-increasing production of NMs, there is a necessity to evaluate the probability of consequential adverse effects, not only in health but also in clinically asymptomatic liver, as part of risk stratification strategies. In this study, two unique disease initiation and maintenance protocols were developed and utilised to mimic steatosis and pre-fibrotic NASH in scaffold-free 3D liver microtissues (MT) composed of primary human hepatocytes, hepatic stellate cells, Kupffer cells and sinusoidal endothelial cells. The characterized diseased MT were utilized for the toxicological assessment of a panel of xenobiotics. Highlights from the study included: 1. Clear experimental evidence for the pre-existing liver disease is important in the augmentation of xenobiotic-induced hepatotoxicity and 2. NMs are able to activate stellate cells. The data demonstrated that pre-existing disease is vital in the intensification of xenobiotic-induced liver damage. Therefore, it is imperative that all stages of the wide spectrum of liver disease are incorporated in risk assessment strategies. This is of significant consequence, as a substantial number of the general population suffer from sub-clinical liver injury without any apparent or diagnosed manifestations.
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Enfermedad del Hígado Graso no Alcohólico , Animales , Células Endoteliales/metabolismo , Hepatocitos , Humanos , Macrófagos del Hígado , Hígado , Enfermedad del Hígado Graso no Alcohólico/inducido químicamente , Enfermedad del Hígado Graso no Alcohólico/metabolismoRESUMEN
The mechanical properties of cells influence their cellular and subcellular functions, including cell adhesion, migration, polarization, and differentiation, as well as organelle organization and trafficking inside the cytoplasm. Yet reported values of cell stiffness and viscosity vary substantially, which suggests differences in how the results of different methods are obtained or analyzed by different groups. To address this issue and illustrate the complementarity of certain approaches, here we present, analyze, and critically compare measurements obtained by means of some of the most widely used methods for cell mechanics: atomic force microscopy, magnetic twisting cytometry, particle-tracking microrheology, parallel-plate rheometry, cell monolayer rheology, and optical stretching. These measurements highlight how elastic and viscous moduli of MCF-7 breast cancer cells can vary 1,000-fold and 100-fold, respectively. We discuss the sources of these variations, including the level of applied mechanical stress, the rate of deformation, the geometry of the probe, the location probed in the cell, and the extracellular microenvironment.
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Análisis de la Célula Individual/métodos , Fenómenos Biomecánicos , Adhesión Celular , Movimiento Celular , Humanos , Dispositivos Laboratorio en un Chip , Células MCF-7 , Estrés MecánicoRESUMEN
Cell-based therapeutics, such as in vitro manufactured red blood cells (mRBCs), are different to traditional biopharmaceutical products (the final product being the cells themselves as opposed to biological molecules such as proteins) and that presents a challenge of developing new robust and economically feasible manufacturing processes, especially for sample purification. Current purification technologies have limited throughput, rely on expensive fluorescent or magnetic immunolabeling with a significant (up to 70%) cell loss and quality impairment. To address this challenge, previously characterized mechanical properties of umbilical cord blood CD34+ cells undergoing in vitro erythropoiesis were used to develop an mRBC purification strategy. The approach consists of two main stages: (a) a microfluidic separation using inertial focusing for deformability-based sorting of enucleated cells (mRBC) from nuclei and nucleated cells resulting in 70% purity and (b) membrane filtration to enhance the purity to 99%. Herein, we propose a new route for high-throughput (processing millions of cells/min and mls of medium/min) purification process for mRBC, leading to high mRBC purity while maintaining cell integrity and no alterations in their global gene expression profile. Further adaption of this separation approach offers a potential route for processing of a wide range of cellular products.
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Separación Celular/instrumentación , Eritrocitos/citología , Filtración/instrumentación , Técnicas Analíticas Microfluídicas/instrumentación , Células Madre/citología , Línea Celular , Diseño de Equipo , HumanosRESUMEN
BACKGROUND: With ever-increasing exposure to engineered nanomaterials (NMs), there is an urgent need to evaluate the probability of consequential adverse effects. The potential for NM translocation to distal organs is a realistic prospect, with the liver being one of the most important target organs. Traditional in vitro or ex vivo hepatic toxicology models are often limiting (i.e. short life-span, reduced metabolic activity, lacking important cell populations, etc.). In this study, we scrutinize a 3D human liver microtissue (MT) model (composed of primary hepatocytes and non-parenchymal cells). This unique experiment benefits from long-term (3 weeks) repeated very low exposure concentrations, as well as incorporation of recovery periods (up to 2 weeks), in an attempt to account for the liver's recovery capacity in vivo. As a means of assessing the toxicological potential of NMs, cell cytotoxicity (cell membrane integrity and aspartate aminotransferase (AST) activity), pro/anti-inflammatory response and hepatic function were investigated. RESULTS: The data showed that 2 weeks of cell culture might be close to limits before subtle ageing effects start to overshadow low sub-lethal NM-induced cellular responses in this test system (adenylate kinase (AK) cytotoxicity assay). We showed that in vitro AST measurement are not suitable in a nanotoxicological context. Moreover, the cytokine analysis (IL6, IL8, IL10 and TNF-α) proved useful in highlighting recovery periods as being sufficient for allowing a reduction in the pro-inflammatory response. Next, low soluble NM-treated MT showed a concentration-dependent penetration of materials deep into the tissue. CONCLUSION: In this study the advantages and pitfalls of the multi-cellular primary liver MT are discussed. Furthermore, we explore a number of important considerations for allowing more meaningful in vitro vs. in vivo comparisons in the field of hepatic nanotoxicology.
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Enfermedad Hepática Inducida por Sustancias y Drogas/etiología , Hepatocitos/efectos de los fármacos , Macrófagos del Hígado/efectos de los fármacos , Hígado/efectos de los fármacos , Nanoestructuras/toxicidad , Técnicas de Cultivo de Tejidos/métodos , Albúminas/metabolismo , Supervivencia Celular/efectos de los fármacos , Técnicas de Cocultivo , Citocinas/metabolismo , Hepatocitos/metabolismo , Hepatocitos/patología , Humanos , Macrófagos del Hígado/metabolismo , Macrófagos del Hígado/patología , Hígado/metabolismo , Hígado/patología , Pruebas de Función HepáticaRESUMEN
BACKGROUND: Surgical treatment of osteochondral lesions of the talus affecting the medial aspect of the talar dome is typically performed using medial malleolar osteotomy to optimize access. This study compares clinical outcomes of lesions repaired using biologic inlay osteochondral reconstruction in patients who did or did not undergo medial malleolar osteotomy, depending on defect dimensions. METHODS: Patients treated for osteochonral lesions of the talus through a medial mallolar approach or arthroscopically-assisted approach were prospectively followed. Assessment tools consisted of the visual analogue scale (VAS) and the American Orthopaedic Foot and Ankle Society Ankle-Hindfoot score (AOFAS). The magnetic resonance observation of cartilage repair tissue (MOCART) score was used postoperatively. RESULTS: Data for 24 patients (mean age 34years, mean follow-up 22 months) was analyzed. Mean preoperative/final AOFAS and VAS in those who underwent osteotomy were 57.7/81.2 and 5.7/1.9 (p<0.001), respectively. In those who underwent arthroscopically-assisted reconstruction, mean preoperative/final AOFAS and VAS were 54.4/84.0 and 7.6/2.0 (p<0.001), respectively. There was no difference in mean MOCART score (p=0.662) for those treated with osteotomy (67.3) compared to those without (70.8). CONCLUSIONS: Osteochondral lesions of the talar dome can be treated successfully by biological inlay osteochondral reconstruction technique without medial malleolar osteotomy, with good to excellent clinical outcomes expected. MRI demonstrates good integration of the graft into surrounding tissue.
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Astrágalo/cirugía , Adolescente , Adulto , Artroscopía , Trasplante Óseo/métodos , Femenino , Humanos , Imagen por Resonancia Magnética , Masculino , Persona de Mediana Edad , Osteotomía/métodos , Procedimientos de Cirugía Plástica , Tibia/cirugía , Trasplante Autólogo , Resultado del Tratamiento , Adulto JovenRESUMEN
Articular cartilage damage to the acetabulum is frequently associated with femoroacetabular impingement, and there are considerable long-term implications for such injury with regard to maintenance of a healthy hip joint and quality of life. Developing treatments capable of restoring articular cartilage to acetabular cartilage defects is of great importance if hip preservation treatments are to be successful. Ideally, such methods should be performed in a minimally invasive manner and be capable of restoring durable repair tissue that reconstitutes a healthy osteochondral unit and that continues to function effectively over the long term.
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Cartílago Articular , Quitosano , Pinzamiento Femoroacetabular , Acetábulo , Estudios de Seguimiento , Articulación de la Cadera , Humanos , Calidad de VidaRESUMEN
PURPOSE: To examine the quality of arthroscopic cartilage debridement using a curette technique by comparing regional and morphologic variations within cartilage lesions prepared in human cadaveric knee specimens for the purpose of cartilage repair procedures. A secondary aim was to compare the histologic properties of cartilage lesions prepared by surgeons of varying experience. METHODS: Standardized cartilage lesions (8 mm × 15 mm), located to the medial/lateral condyle and medial/lateral trochlea were created within 12 human cadaver knees by 40 orthopaedic surgeons. Participants were instructed to create full-thickness cartilage defects within the marked area, shouldered by uninjured vertical walls of cartilage, and to remove the calcified cartilage layer, without violating the subchondral plate. Histologic specimens were prepared to examine the verticality of surrounding cartilage walls at the front and rear aspects of the lesions, and to characterize the properties of the surrounding cartilage, the cartilage wall profile, the debrided lesion depth, bone sinusoid access, and the bone surface profile. Comparative analysis of cartilage wall verticality measured as deviation from perpendicular was performed, and Spearman's rank correlation analysis was used to examine associations between debrided wall verticality and surgeon experience. RESULTS: Mean cartilage wall verticality relative to the base of the lesion was superior at the rear aspect of the lesion compared to the front aspect (12.9° vs 29.2°, P < .001). Variability was identified in the morphology of the surrounding cartilage (P < .001), cartilage wall profile (P = .016), debrided lesion depth (P = .028), bone surface profile (P = .040), and bone sinusoid access (P = .009), with sinusoid access identified in 42% of cases. There was no significant association of cartilage lesion wall verticality and surgeon years in practice (rs = 0.161, P = .065) or arthroscopic caseload (rs = -0.071, P = .419). CONCLUSIONS: Arthroscopic cartilage lesion preparation using standard curette technique in a human cadaveric knee model results in inferior perpendicularity of the surrounding cartilage walls at the front aspect of the defect, compared to the rear aspect. This technique has shown significant variability in the depth of debridement, with debridement depths identified as either too superficial or too deep to the calcified cartilage layer in more than 60% of cases in this study. Surgeon experience does not appear to impact the morphologic properties of cartilage lesions prepared arthroscopically using ring curettes. CLINICAL RELEVANCE: To optimize restoration of hyaline-like cartilage tissue, careful attention to prepared cartilage lesion morphology is advised when arthroscopically performing cartilage repair, given the tendency for standard curette technique to create inferior verticality of cartilage walls at the front of the lesion, and the variable depth of debridement achieved.
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Artroscopía/métodos , Enfermedades de los Cartílagos/cirugía , Cartílago Articular/cirugía , Desbridamiento/métodos , Cartílago Hialino/cirugía , Articulación de la Rodilla/cirugía , Adulto , Cadáver , Enfermedades de los Cartílagos/patología , Cartílago Articular/patología , Competencia Clínica , Legrado/instrumentación , Femenino , Humanos , Cartílago Hialino/patología , Articulación de la Rodilla/patología , Masculino , Cirujanos Ortopédicos , Encuestas y CuestionariosRESUMEN
We describe a quantitative, high-precision, high-throughput method for measuring the mechanical properties of cells in suspension with a microfluidic device, and for relating cell mechanical responses to protein expression levels. Using a high-speed (750 fps) charge-coupled device camera, we measure the driving pressure Δp, maximum cell deformation εmax, and entry time tentry of cells in an array of microconstrictions. From these measurements, we estimate population averages of elastic modulus E and fluidity ß (the power-law exponent of the cell deformation in response to a step change in pressure). We find that cell elasticity increases with increasing strain εmax according to E ⼠εmax, and with increasing pressure according to E â¼ Δp. Variable cell stress due to driving pressure fluctuations and variable cell strain due to cell size fluctuations therefore cause significant variability between measurements. To reduce measurement variability, we use a histogram matching method that selects and analyzes only those cells from different measurements that have experienced the same pressure and strain. With this method, we investigate the influence of measurement parameters on the resulting cell elastic modulus and fluidity. We find a small but significant softening of cells with increasing time after cell harvesting. Cells harvested from confluent cultures are softer compared to cells harvested from subconfluent cultures. Moreover, cell elastic modulus increases with decreasing concentration of the adhesion-reducing surfactant pluronic. Lastly, we simultaneously measure cell mechanics and fluorescence signals of cells that overexpress the GFP-tagged nuclear envelope protein lamin A. We find a dose-dependent increase in cell elastic modulus and decrease in cell fluidity with increasing lamin A levels. Together, our findings demonstrate that histogram matching of pressure, strain, and protein expression levels greatly reduces the variability between measurements and enables us to reproducibly detect small differences in cell mechanics.
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Células/metabolismo , Microtecnología/métodos , Fenómenos Biomecánicos , Células Cultivadas , Humanos , Células K562 , Dispositivos Laboratorio en un Chip , Poloxámero/farmacología , Reproducibilidad de los Resultados , Estrés Mecánico , Factores de Tiempo , Tripsina/metabolismoRESUMEN
Microfluidic microdroplets have increasingly found application in biomolecular sensing as well as nanomaterials growth. More recently the synthesis of plasmonic nanostructures in microdroplets has led to surface-enhanced Raman spectroscopy (SERS)-based sensing applications. However, the study of nanoassembly in microdroplets has previously been hindered by the lack of on-chip characterization tools, particularly at early timescales. Enabled by a refractive index matching microdroplet formulation, dark-field spectroscopy is exploited to directly track the formation of nanometer-spaced gold nanoparticle assemblies in microdroplets. Measurements in flow provide millisecond time resolution through the assembly process, allowing identification of a regime where dimer formation dominates the dark-field scattering and SERS. Furthermore, it is shown that small numbers of nanoparticles can be isolated in microdroplets, paving the way for simple high-yield assembly, isolation, and sorting of few nanoparticle structures.
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Nanopartículas del Metal/química , Microfluídica/métodos , Fenómenos Ópticos , Espectrometría Raman/métodos , Oro/química , RefractometríaRESUMEN
PURPOSE: To evaluate the fixation integrity at time zero of a type I/III collagen patch secured to a chondral defect in the porcine knee using methods typically employed in autologous chondrocyte implantation (ACI) and matrix-assisted chondrocyte implantation. METHODS: Twenty-four porcine knee specimens underwent a medial parapatellar arthrotomy. A prefabricated template was used to create cartilage defects of 2 cm(2) in the medial femoral condyle. A size-matched collagen patch was fashioned. Four methods of fixation to the chondral defect were analyzed: group 1-saline, group 2-fibrin glue around the periphery of the patch, group 3-fibrin glue applied to the base of the defect and around the periphery of the patch, group 4-6-0 vicryl suture and fibrin glue around the periphery of the patch. Collagen patch fixation was assessed at intervals of 60, 300, 600, 900, and 1,200 cycles from full extension to 90° of flexion, performed manually without application of axial force. Patch fixation was evaluated by 2 independent observers using a customized scoring scale. RESULTS: Mean peripheral detachment of the patch and chondral defect uncovering remained less than 25% for all groups. Area of defect uncovering was significantly increased in group 2 compared with group 4 after 900 and 1,200 cycles (P = .0014 and P = .0025, respectively). Fibrin glue applied to the base of the defect, or suturing of the patch, reduced deformation significantly after 900 cycles. CONCLUSIONS: Suture increases the stability of fixation of a type I/III collagen patch to a chondral defect better than fibrin glue alone in the porcine knee after repetitive cycling, with respect to patch detachment and chondral defect uncovering. Application of fibrin glue to the base of the defect, or securing the patch with suture, decreases collagen patch deformation. CLINICAL RELEVANCE: In cases where minimally invasive techniques do not allow suture fixation of the collagen patch, scaffold fixation may be compromised during articular motion protocols typically used after second- and third-generation ACI procedures.
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Colágeno , Adhesivo de Tejido de Fibrina , Rodilla de Cuadrúpedos/cirugía , Suturas , Adhesivos Tisulares , Andamios del Tejido , Animales , Modelos Animales , PorcinosRESUMEN
We describe a method for quantifying the mechanical properties of cells in suspension with a microfluidic device consisting of a parallel array of micron-sized constrictions. Using a high-speed charge-coupled device camera, we measure the flow speed, cell deformation, and entry time into the constrictions of several hundred cells per minute during their passage through the device. From the flow speed and the occupation state of the microconstriction array with cells, the driving pressure across each constriction is continuously computed. Cell entry times into microconstrictions decrease with increased driving pressure and decreased cell size according to a power law. From this power-law relationship, the cell elasticity and fluidity can be estimated. When cells are treated with drugs that depolymerize or stabilize the cytoskeleton or the nucleus, elasticity and fluidity data from all treatments collapse onto a master curve. Power-law rheology and collapse onto a master curve are predicted by the theory of soft glassy materials and have been previously shown to describe the mechanical behavior of cells adhering to a substrate. Our finding that this theory also applies to cells in suspension provides the foundation for a quantitative high-throughput measurement of cell mechanical properties with microfluidic devices.
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Fenómenos Fisiológicos Celulares , Dispositivos Laboratorio en un Chip , Técnicas Analíticas Microfluídicas/instrumentación , Línea Celular Tumoral , Núcleo Celular/efectos de los fármacos , Núcleo Celular/fisiología , Fenómenos Fisiológicos Celulares/efectos de los fármacos , Citoesqueleto/efectos de los fármacos , Citoesqueleto/fisiología , Diseño de Equipo , Vidrio/química , Humanos , Fenómenos Mecánicos , Técnicas Analíticas Microfluídicas/métodos , Microtecnología/instrumentación , Modelos Teóricos , Presión , ReologíaRESUMEN
Phospholipid vesicles are common model systems for cell membranes. Important aspects of the membrane function relate to its mechanical properties. Here we have investigated the deformation behaviour of phospholipid vesicles in a dual-beam laser trap, also called an optical stretcher. This study explicitly makes use of the inherent heating present in such traps to investigate the dependence of vesicle deformation on temperature. By using lasers with different wavelengths, optically induced mechanical stresses and temperature increase can be tuned fairly independently with a single setup. The phase transition temperature of vesicles can be clearly identified by an increase in deformation. In the case of no heating effects, a minimal model for drop deformation in an optical stretcher and a more specific model for vesicle deformation that takes explicitly into account the angular dependence of the optical stress are presented to account for the experimental results. Elastic constants are extracted from the fitting procedures, which agree with literature data. This study demonstrates the utility of optical stretching, which is easily combined with microfluidic delivery, for the future serial, high-throughput study of the mechanical and thermodynamic properties of phospholipid vesicles.
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Membrana Celular/química , Microfluídica , Fosfolípidos/química , Elasticidad , Rayos Láser , Modelos Teóricos , Óptica y Fotónica , Temperatura , TermodinámicaRESUMEN
Using devices with microfluidic channels can allow for precise control over liquids flowing through them. Merging flows of immiscible liquids can create emulsions with highly monodispersed microdroplets within a carrier liquid, which are ideal for miniaturised reaction vessels which can be generated with a high throughput of tens of thousands of droplets per second. Control of the size and composition of these droplets is generally performed by controlling the pumping system pushing the liquids into the device; however, this is an indirect manipulation and inadequate if absolute precision is required in the size or composition of the droplets. In this work, we extend the previous development of image-based closed-loop feedback control over microdroplet generation to allow for the control of not only the size of droplets but also the composition by merging two aqueous flows. The feedback allows direct control over the desired parameters of volume and ratio of the two components over a wide range of ratios and outperforms current techniques in terms of monodispersity in volume and composition. This technique is ideal for situations where precise control over droplets is critical, or where a library of droplets of different concentrations but the same volume is required.
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We present two electromagnetic frameworks to compare the surface stresses on spheroidal particles in the optical stretcher (a dual-beam laser trap that can be used to capture and deform biological cells). The first model is based on geometrical optics (GO) and limited in its applicability to particles that are much greater than the incident wavelength. The second framework is more sophisticated and hinges on the generalized Lorenz-Mie theory (GLMT). Despite the difference in complexity between both theories, the stress profiles computed with GO and GLMT are in good agreement with each other (relative errors are on the order of 1-10%). Both models predict a diminishing of the stresses for larger wavelengths and a strong increase of the stresses for shorter laser-cell distances. Results indicate that surface stresses on a spheroid with an aspect ratio of 1.2 hardly differ from the stresses on a sphere of similar size. Knowledge of the surface stresses and whether or not they redistribute during the stretching process is of crucial importance in real-time applications of the stretcher that aim to discern the viscoelastic properties of cells for purposes of cell characterization, sorting, and medical diagnostics.
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Algoritmos , Microesferas , Modelos Teóricos , Pinzas Ópticas , Simulación por Computador , Dosis de Radiación , Estrés MecánicoRESUMEN
There has been an increasing drive toward miniaturizing and accelerating experiments with droplet-based microfluidics across the chemical disciplines. Current applications take advantage of the numerous techniques for manipulating nano- to femtoliter droplets within microfluidic devices. To expand the range of possible applications, we have developed a method for compartmentalizing pure compounds within droplets, at a gradient of concentrations, starting from chemical mixtures. In this technique, a mixture is injected into an ultra performance liquid chromatography (UPLC) system, and droplets are generated from the LC output at a frequency high enough to fraction each compound into approximately 10(5) droplets, compartmentalizing pure compounds into a sequence of droplets with a range of concentrations spanning 2-3 orders of magnitude. Here we used fluorescent dyes to quantify the concentration profile of the droplet collections, and to demonstrate the correspondence between the concentration profile of the droplets and the compound elution profile monitored with a UV absorbance detector, allowing the use of compounds that are not fluorescently labeled but show UV absorbance. Hence this technique is applicable to a wide variety of applications that require both compound purity and the ability to probe a variety of concentrations, such as drug screening and titrations.
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Microfluídica , Cromatografía Líquida de Alta Presión , Electroquímica , Colorantes Fluorescentes/química , Microfluídica/métodos , NanotecnologíaRESUMEN
Massive and retracted tears of the supraspinatus and infraspinatus tendons of the rotator cuff are associated with great pain and disability and may be considered "non-repairable," depending on the extent of injury and the experience of the treating clinician. The technique of anatomic vector repair of the rotator cuff is a surgical treatment method that enables the surgeon to accurately characterize the injury pattern and successfully repair many of these debilitating injuries anatomically in a stepwise manner, often in cases that would have otherwise been treated with a less preferable surgical procedure that does not restore native anatomy.
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The recent boom in single-cell omics has brought researchers one step closer to understanding the biological mechanisms associated with cell heterogeneity. Rare cells that have historically been obscured by bulk measurement techniques are being studied by single cell analysis and providing valuable insight into cell function. To support this progress, novel upstream capabilities are required for single cell preparation for analysis. Presented here is a droplet microfluidic, image-based single-cell sorting technique that is flexible and programmable. The automated system performs real-time dual-camera imaging (brightfield & fluorescent), processing, decision making and sorting verification. To demonstrate capabilities, the system was used to overcome the Poisson loading problem by sorting for droplets containing a single red blood cell with 85% purity. Furthermore, fluorescent imaging and machine learning was used to load single K562 cells amongst clusters based on their instantaneous size and circularity. The presented system aspires to replace manual cell handling techniques by translating expert knowledge into cell sorting automation via machine learning algorithms. This powerful technique finds application in the enrichment of single cells based on their micrographs for further downstream processing and analysis.
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Técnicas Analíticas Microfluídicas/métodos , Reconocimiento de Normas Patrones Automatizadas/métodos , Análisis de la Célula Individual/métodos , Separación Celular , Toma de Decisiones , Citometría de Flujo , Humanos , Células K562 , Aprendizaje AutomáticoRESUMEN
Cell sorting and isolation from a heterogeneous mixture is a crucial task in many aspects of cell biology, biotechnology and medicine. Recently, there has been an interest in methods allowing cell separation upon their intrinsic properties such as cell size and deformability, without the need for use of biochemical labels. Inertial focusing in spiral microchannels has been recognised as an attractive approach for high-throughput cell sorting for myriad point of care and clinical diagnostics. Particles of different sizes interact to a different degree with the fluid flow pattern generated within the spiral microchannel and that leads to particles ordering and separation based on size. However, the deformable nature of cells adds complexity to their ordering within the spiral channels. Herein, an additional force, deformability-induced lift force (FD), involved in the cell focusing mechanism within spiral microchannels has been identified, investigated and reported for the first time, using a cellular deformability model (where the deformability of cells is gradually altered using chemical treatments). Using this model, we demonstrated that spiral microchannels are capable of separating cells of the same size but different deformability properties, extending the capability of the previous method. We have developed a unique label-free approach for deformability-based purification through coupling the effect of FD with inertial focusing in spiral microchannels. This microfluidic-based purification strategy, free of the modifying immuno-labels, allowing cell processing at a large scale (millions of cells per min and mls of medium per minute), up to high purities and separation efficiency and without compromising cell quality.
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Separación Celular , Citofotometría , Dispositivos Laboratorio en un Chip , Técnicas Analíticas Microfluídicas , Células Cultivadas , Citofotometría/instrumentación , Humanos , Células Jurkat , Técnicas Analíticas Microfluídicas/instrumentación , Tamaño de la Partícula , Propiedades de SuperficieRESUMEN
Correction for 'Deformability-induced lift force in spiral microchannels for cell separation' by Ewa Guzniczak et al., Lab Chip, 2020, 20, 614-625.
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The thin and flexible nature of optical fibres often makes them the ideal technology to view biological processes in-vivo, but current microendoscopic approaches are limited in spatial resolution. Here, we demonstrate a route to high resolution microendoscopy using a multicore fibre (MCF) with an adiabatic multimode-to-single-mode "photonic lantern" transition formed at the distal end by tapering. We show that distinct multimode patterns of light can be projected from the output of the lantern by individually exciting the single-mode MCF cores, and that these patterns are highly stable to fibre movement. This capability is then exploited to demonstrate a form of single-pixel imaging, where a single pixel detector is used to detect the fraction of light transmitted through the object for each multimode pattern. A custom computational imaging algorithm we call SARA-COIL is used to reconstruct the object using only the pre-measured multimode patterns themselves and the detector signals.