A systematic review on the culture methods and applications of 3D tumoroids for cancer research and personalized medicine.

Cancer is a highly heterogeneous disease, and thus treatment responses vary greatly between patients. To improve therapy efficacy and outcome for cancer patients, more representative and patient-specific preclinical models are needed. Organoids and tumoroids are 3D cell culture models that typically retain the genetic and epigenetic characteristics, as well as the morphology, of their tissue of origin. Thus, they can be used to understand the underlying mechanisms of cancer initiation, progression, and metastasis in a more physiological setting. Additionally, co-culture methods of tumoroids and cancer-associated cells can help to understand the interplay between a tumor and its tumor microenvironment. In recent years, tumoroids have already helped to refine treatments and to identify new targets for cancer therapy. Advanced culturing systems such as chip-based fluidic devices and bioprinting methods in combination with tumoroids have been used for high-throughput applications for personalized medicine. Even though organoid and tumoroid models are complex in vitro systems, validation of results in vivo is still the common practice. Here, we describe how both animal- and human-derived tumoroids have helped to identify novel vulnerabilities for cancer treatment in recent years, and how they are currently used for precision medicine.

3D-printed stereolithographic fluidic devices for automatic nonsupported microelectromembrane extraction and clean-up of wastewater samples.

BACKGROUND: There is a quest of novel functional and reliable platforms for enhancing the efficiency of microextraction approaches in troublesome matrices, such as industrial wastewaters. 3D printing has been proven superb in the analytical field to act as the springboard of microscale extraction approaches. RESULTS: In this work, low-force stereolithography (SL) was exploited for 3D printing and prototyping bespoke fluidic devices for accommodating nonsupported microelectromembrane extraction (µEME). The analytical performance of 3D-printed µEME devices with distinct cross-sections, including square, circle, and obround, and various channel dimensions was explored against that of commonly used circular polytetrafluoroethylene (PTFE) tubing in flow injection systems. A computer-controlled millifluidic system was harnessed for the (i) automatic liquid-handling of minute volumes of donor, acceptor, and organic phases at the low µL level that spanned from 3 to 44 µL in this work, (ii) formation of three-phase µEME, (iii) in-line extraction, (iv) flow-through optical detection of the acceptor phase, and (v) solvent removal and regeneration of the µEME device and fluidic lines. Using methylene blue (MB) as a model analyte, experimental results evinced that the 3D-printed channels with an obround cross-section (2.5 mm × 2.5 mm) were the most efficient in terms of absolute extraction recovery (59%), as compared to PTFE tubing of 2.5 mm inner diameter (27%). This is attributed to the distinctive convex interface of the organic phase (1-octanol), with a more pronounced laminar pattern, in 3D-printed SL methacrylate-based fluidic channels against that of PTFE tubing on account of the enhanced 1-octanol wettability and lower contact angles for the 3D-printed devices. The devices with obround channels were leveraged for the automatic µEME and in-line clean-up of MB in high matrix textile dyeing wastewater samples with relative recoveries ≥81%, RSD% ≤ 17.1% and LOD of 1.3 mg L-1. The 3D-printed nonsupported µEME device was proven superb for the analysis of wastewater samples with an elevated ionic strength (0.7 mol L-1 NaCl, 5000 mg L-1 Na2CO3, and 0.013 mol L-1 NaOH) with recorded electric currents below 12 µA. NOVELTY: The coupling of 3D printing with nonsupported µEME in automatic flow-based systems is herein proposed for the first time and demonstrated for the clean-up of troublesome samples, such as wastewaters.

Artificial Monovalent Metal Ion-Selective Fluidic Devices Based on Crown Ether@Metal-Organic Frameworks with Subnanochannels.

Precise regulation of ion transport through nanoscale pores will profoundly impact diverse fields from separation to energy conversion but is still challenging to achieve in artificial ion channels. Herein, inspired by the exquisite ion selectivity of biological Na+ channels, we have successfully fabricated hierarchically grown metal-organic frameworks (MOFs) on an asymmetrical substrate assisted by atomically thin nanoporous graphene. Efficient separation of monovalent metal ions is realized by encapsulating 18-crown-6 into MOF crystals. The resulting 18-crown-6@ZIF-67/ZIF-8 device, with subnanochannels and specific K+ binding sites, shows an ultrahigh Li+ conductivity of 1.46 × 10-2 S cm-1 and selectivities of 9.56 and 6.43 for Li+/K+ and Na+/K+, respectively. The Li+ conductivity is around 1-2 orders of magnitude higher than reported values for the other MOF materials. It is the first time that MOFs with subnanochannels realize selective transport of Na+ (ionic diameter of 1.9 Å) over K+ (2.6 Å) based on subangstrom differences in their ionic diameter. Our work opens new avenues to develop crown ether@MOF platforms toward efficient ion transistors, fluidic logic devices, and biosensors.

3D printing for the integration of porous materials into miniaturised fluidic devices: A review.

Porous materials facilitate the efficient separation of chemicals and particulate matter by providing selectivity through structural and surface properties and are attractive as sorbent owing to their large surface area. This broad applicability of porous materials makes the integration of porous materials and microfluidic devices important in the development of more efficient, advanced separation platforms. Additive manufacturing approaches are fundamentally different to traditional manufacturing methods, providing unique opportunities in the fabrication of fluidic devices. The complementary 3D printing (3DP) methods are each accompanied by unique opportunities and limitations in terms of minimum channel size, scalability, functional integration and automation. This review focuses on the developments in the fabrication of 3DP miniaturised fluidic devices with integrated porous materials, focusing polymer-based methods including fused filament fabrication (FFF), inkjet 3D printing and digital light projection (DLP). The 3DP methods are compared based on resolution, scope for multimaterial printing and scalability for manufacturing. As opportunities for printing pores are limited by resolution, the focus is on approaches to incorporate materials with sub-micron pores to be used as membrane, sorbent or stationary phase in separation science using Post-Print, Print-Pause-Print and In-Print processes. Technical aspects analysing the efficiency of the fabrication process towards scalable manufacturing are combined with application aspects evaluating the separation and/or extraction performance. The review is concluded with an overview on achievements and opportunities for manufacturable 3D printed membrane/sorbent integrated fluidic devices.

Scalable 3D printing method for the manufacture of single-material fluidic devices with integrated filter for point of collection colourimetric analysis.

Assembly and bonding are major obstacles in manufacturing of functionally integrated fluidic devices. Here we demonstrate a single-material 3D printed device with an integrated porous structure capable of filtering particulate matter for the colourimetric detection of iron from soil and natural waters. Selecting a PolyJet 3D printer for its throughput, integrated filters were created exploiting a phenomenon occurring at the interface between the commercially available build material (Veroclear-RGD810) and water-soluble support material (SUP707). The porous properties were tuneable by varying the orientation of the print head relative to the channel and by varying the width of the build material. Porous structures ranging from 100 to 200 µm in thickness separated the sample and reagent chambers, filtering particles larger than 15 µm in diameter. Maintaining the manufacturing throughput of the Polyjet printer, 221 devices could be printed in 1.5 h (∼25 s per device). Including the 12 h post-processing soak in sodium hydroxide to remove the solid support material, the total time to print and process 221 devices was 13.5 h (3.6 min per device), with a material cost of $2.50 each. The applicability of the fluidic device for point of collection analysis was evaluated using colourimetric determination of iron from soil slurry and environmental samples. Following the reduction of Fe3+ to Fe2+ using hydroxylammonium chloride, samples were introduced to the fluidic device where particulate matter was retained by the filter, allowing for particulate-free imaging of the red complex formed with 1,10-phenanthroline using a smartphone camera. The calibration curve ranged from of 1-100 mg L-1 Fe2+ and good agreement (95%) was obtained between the point of collection device and Sector Field ICP-MS.

Rectification of Mobile Leidenfrost Droplets by Planar Ratchets.

The self-transportation of mobile Leidenfrost droplets with well-defined direction and velocity on millimetric ratchets is one of the most representative and spectacular phenomena in droplet dynamics. Despite extensive progress in the ability to control the spatiotemporal propagation of droplets, it remains elusive how the individual ratchet units, as well as the interactions within their arrays, are translated into the collective droplet dynamics. Here, simple planar ratchets characterized by uniform height normal to the surface are designed. It is revealed that on planar ratchets, the transport dynamics of Leidenfrost droplets is dependent not only on individual units, but also on the elegant coordination within their arrays dictated by their topography. The design of planar ratchets enriches the fundamental understanding of how the surface topography is translated into dynamic and collective droplet transport behaviors, and also imparts higher applicability in microelectromechanical system based fluidic devices.

Chips: How to build and implement fluidic devices in flow based systems.

The development of automatic analyzers based on flow techniques involves the use and continuous innovation of fluidic devices. New trends tend toward miniaturization of sophisticated fluidic platforms requiring continuous advances in this field. The availability of a mechanic and electronic workshop together with the know-how to build new fluidic devices provides the tools for the creation of innovative instrumentation and stimulates the creativity of analytical chemists. Thus, in this review we present how to build and use flow-based fluidic devices, together with the tools required, such as computerized controlled lathes, milling machines, laser engraver machines, low-temperature co-fired ceramics technology and 3D printers, highlighting their strong and weak points. In addition, some flow based methods exploiting innovative fluidic platforms are also presented as a way of example of the possible devices these tools can provide and their potential applications.

Metalless electrodes for capacitively coupled contactless conductivity detection on electrophoresis microchips.

This paper describes the use of ionic solutions as sensing electrodes for capacitively coupled contactless conductivity detection on electrophoresis microchips. Initially, two channels were engraved in a PMMA holder by using a CO2 laser system and sealed with a thin adhesive membrane. PDMS electrophoresis chips were fabricated by soft lithography and reversibly sealed against the polymer membrane. Different ionic solutions were investigated as metalless electrodes. The electrode channels were filled with KCl solutions prepared in conductivity values from approximately 10 to 40 S/m. The best analytical response was achieved using the KCl solution with 21.9 S/m conductivity (2 mol/L). Besides KCl, we also tested NaCl and LiCl solutions for actuating as detection electrodes. Taking into account the same electrolyte concentration (2 mol/L), the best response was recorded with KCl solution due to its higher ionic conductivity. The optimum operating frequency (400 kHz) and the best sensing electrode (2 mol/L KCl) were used to monitor electrophoretic separations of a mixture containing K(+) , Na(+) , and Li(+) . The use of liquid solutions as sensing electrodes for capacitively coupled contactless conductivity detection measurements has revealed great performance to monitor separations on chip-based devices, avoiding complicated fabrication schemes to include metal deposition and encapsulation of electrodes. The LOD values were estimated to be 28, 40, and 58 µmol/L for K(+) , Na(+) , and Li(+) , respectively, what is comparable to that of conventional metal electrodes. When compared to the use metal electrodes, the proposed approach offers advantages regarding the easiness of fabrication, simplicity, and lower cost per device.

Characterization of microchip electrophoresis devices fabricated by direct-printing process with colored toner.

This paper reports for the first time the use of colored toner to produce polyester toner (PT) ME devices. Colored PT devices were designed in drawing software and printed on a polyester film using a color laser printer with 3600 dpi resolution. The colored toner is composed of a copolymer mixture (styrene and acrylate), wax, silicon dioxide, and pigments. The presence of silica in the toner composition has enhanced the EOF magnitude and improved the analytical performance. For a pH range between 2 and 12, the EOF measured on a magenta PT chip, for example, ranged from 3.8 to 5.8 (× 10(-4) cm(2) V(-1) s(-1) ). Typical separations of inorganic cations (K(+) , Na(+) , and Li(+) ) were used as model system to investigate the analytical feasibility of the proposed devices. The repeatability for the migration times of all analytes exhibited RSD values lower than 1% (n = 10). The separation efficiencies found on colored PT devices ranged from 10 000 to 49 000 plates/m, which means between 7 and 23% of the maximum theoretical efficiency on this microfluidic platform (1.85 × 10(5) plates/m). The improvements achieved on the proposed devices are associated with the small additional amount of silica on the toner composition as well as the printing of channels with smoother surfaces and better uniformity when compared to the conventional PT chips printed with monochromatic laser printers.

Droplet-based microreactors for continuous production of palladium nanocrystals with controlled sizes and shapes.

Droplet-based microreactors are used for the continuous production of Pd nanocrystals. Specifically, commercially available polytetrafluoroethylene (PTFE) tube and silica capillaries are utilized to fabricate a fluidic device capable of generating water-in-oil droplets. In addition to the feasibility of using such droplets as microreactors for conducting a synthesis, the ability to control the composition and concentration of reagents by adjusting the flow rates is demonstrated; reagents are mixed by periodically pinching the PTFE tube, and nanocrystals are produced with uniform size distribution in a continuous fashion. The capability to tailor the size and shape of the resultant nanocrystals is further demonstrated by introducing the reducing agent and capping agent at different flow rates to control the nucleation and growth processes. The ability to transform a bulk synthesis into a droplet-based system holds great potential for the development of a new route to the high-volume production of nanocrystals.

Evaluation of Superficial and Dimensional Quality Features in Metallic Micro-Channels Manufactured by Micro-End-Milling.

Miniaturization encourages the development of new manufacturing processes capable of fabricating features, like micro-channels, in order to use them for different applications, such as in fuel cells, heat exchangers, microfluidic devices and micro-electromechanical systems (MEMS). Many studies have been conducted on heat and fluid transfer in micro-channels, and they appeared significantly deviated from conventional theory, due to measurement errors and fabrication methods. The present research, in order to deal with this opportunity, is focused on a set of experiments in the micro-milling of channels made of aluminum, titanium alloys and stainless steel, varying parameters, such as spindle speed, depth of cut per pass (ap), channel depth (d), feed per tooth (fz) and coolant application. The experimental results were analyzed in terms of dimensional error, channel profile shape deviation from rectangular and surface quality (burr and roughness). The micro-milling process was capable of offering quality features required on the micro-channeled devices. Critical phenomena, like run-out, ploughing, minimum chip thickness and tool wear, were encountered as an explanation for the deviations in shape and for the surface quality of the micro-channels. The application of coolant and a low depth of cut per pass were significant to obtain better superficial quality features and a smaller dimensional error. In conclusion, the integration of superficial and geometrical features on the study of the quality of micro-channeled devices made of different metallic materials contributes to the understanding of the impact of calibrated cutting conditions in MEMS applications.