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Abstract Three-dimensional (3D) microneedle arrays (MAs) have shown remarkable performances for a wide range of biomedical applications. Achieving advanced customizable 3D MAs for personalized research and treatment remain a formidable challenge. In this paper, we have developed a high-resolution electrohydrodynamic (EHD) 3D printing process for fabricating customizable 3D MAs with economical and biocompatible molten alloy. The critical printing parameters (i.e., voltage and pressure) on the printing process for both two-dimensional (2D) and 3D features are characterized, and an optimal set of printing parameters was obtained for printing 3D MAs. We have also studied the effect of the tip-nozzle separation speed on the final tip dimension, which will directly influence MAs' insertion performance and functions. With the optimal process parameters, we successfully EHD printed customizable 3D MAs with varying spacing distances and shank heights. A 3 × 3 customized 3D MAs configuration with various heights ranging from 0.8 mm to 1 mm and a spacing distance as small as 350 μm were successfully fabricated, in which the diameter of each individual microneedle was as small as 100 μm. A series of tests were conducted to evaluate the printed 3D MAs. The experimental results demonstrated that the printed 3D MAs exhibit good mechanical strength for implanting and good electrical properties for electrophysiological sensing and stimulation. All results show the potential applications of the EHD printing technique in fabricating cost-effective, customizable, high-performance MAs for biomedical applications. 

Abstract While moderately elevated ambient temperatures do not trigger stress responses in plants, they do substantially stimulate the growth of specific organs through a process known as thermomorphogenesis. The basic helix–loop–helix transcription factor PHYTOCHROME-INTERACTING FACTOR 4 (PIF4) plays a central role in regulating thermomorphogenetic hypocotyl elongation in various plant species, including Arabidopsis (Arabidopsis thaliana). Although it is well known that PIF4 and its co-activator HEMERA (HMR) promote plant thermosensory growth by activating genes involved in the biosynthesis and signaling of the phytohormone auxin, the detailed molecular mechanism of such transcriptional activation is not clear. In this report, we investigated the role of the Mediator complex in the PIF4/HMR-mediated thermoresponsive gene expression. Through the characterization of various mutants of the Mediator complex, a tail subunit named MED14 was identified as an essential factor for thermomorphogenetic hypocotyl growth. MED14 was required for the thermal induction of PIF4 target genes but had a marginal effect on the levels of PIF4 and HMR. Further transcriptomic analyses confirmed that the expression of numerous PIF4/HMR-dependent, auxin-related genes required MED14 at warm temperatures. Moreover, PIF4 and HMR physically interacted with MED14 and both were indispensable for the association of MED14 with the promoters of these thermoresponsive genes. While PIF4 did not regulate MED14 levels, HMR was required for the transcript abundance of MED14. Taken together, these results unveil an important thermomorphogenetic mechanism, in which PIF4 and HMR recruit the Mediator complex to activate auxin-related growth-promoting genes when plants sense moderate increases in ambient temperature. 

Temperature control is crucial for live cell imaging, particularly in studies involving plant responses to high ambient temperatures and thermal stress. This study presents the design, development, and testing of two cost-effective heating devices tailored for confocal microscopy applications: an aluminum heat plate and a wireless mini-heater. The aluminum heat plate, engineered to integrate seamlessly with the standard 160 mm × 110 mm microscope stage, supports temperatures up to 36°C, suitable for studies in the range of non-stressful warm temperatures (e.g., 25-27°C forArabidopsis thaliana) and moderate heat stress (e.g., 30-36°C forA. thaliana). We also developed a wireless mini-heater that offers rapid, precise heating directly at the sample slide, with a temperature increase rate over 30 times faster than the heat plate. The wireless heater effectively maintained target temperatures up to 50°C, ideal for investigating severe heat stress and heat shock responses in plants. Both devices performed well in controlled studies, including the real-time analysis of heat shock protein accumulation and stress granule formation inA. thaliana. Our designs are effective and affordable, with total construction costs lower than $300. This accessibility makes them particularly valuable for small laboratories with limited funding. Future improvements could include enhanced heat uniformity, humidity control to mitigate evaporation, and more robust thermal management to minimize focus drift during extended imaging sessions. These modifications would further solidify the utility of our heating devices in live cell imaging, offering researchers reliable, budget-friendly tools for exploring plant thermal biology. 

Electrohydrodynamic (EHD) printing has become a promising and cost-effective technique for producing high-resolution and large-scale features. One widely recognized obstacle in EHD printing is nozzle clogging due to solvent evaporation or ink polymerization. Moreover, printing highly viscous materials often requires pressure or other external force to assist the ink flow during the printing, which increases the complexity of process control and the required energy. In this work, we developed a novel ultrasonic vibration-assisted EHD printhead and associated process to effectively eliminate the nozzle clogging for the printing of high-viscosity and high-evaporation-rate inks. A series of experimental tests were conducted to characterize the printhead design and process parameters (i.e., vibration frequency, vibration amplitude, and printing voltage). The results demonstrated that superimposing ultrasonic vibration on the EHD printing nozzle can effectively enhance current EHD printing capabilities, such as reducing required pressure, eliminating nozzle clogging, and providing stable and continuous printing for high viscosity and high solvent evaporation rate material. With the optimal parameters, a filament with a diameter of around 1 µm can be continuously printed. In the paper, we successfully applied this developed ultrasonic-assisted EHD process to print high-resolution 2D patterns. 

Microheaters have drawn extensive attention for their substantial applications in thermotherapy, gas sensors, thin film preparation, biological research, etc. In plant physiology, uncovering the mechanisms by which plants sense and respond to environmental temperature fluctuations will help us better understand the impact of climate change on crop yield and ecosystem resilience. Currently, microheaters with long-term heating capability have rarely been applied to investigate plant thermal responses. In this study, we applied a direct writing technique to fabricate microheaters suitable for studying plant thermal biology with silver conductive ink. The optimal printing conditions and the heating performance (e.g., stability, durability, reusability) of the printed heaters were thoroughly characterized. The printed microheaters can provide stable and constant heating to plant organs for over four days. When placed near plant leaves to create localized heating, the microheater could successfully activate the expression of a thermoresponsive marker gene in plants. These results demonstrate the potential of applying printed microheaters to study plant thermal biology at the organ and tissue level.