Assembly Yield Research Articles

Modeling reveals the strength of weak interactions in stacked-ring assembly

Cells employ many large macromolecular machines for the execution and regulation of processes that are vital for cell and organismal viability. Interestingly, cells cannot synthesize these machines as functioning units. Instead, cells synthesize the molecular parts that must then assemble into the functional complex. Many important machines, including chaperones such as GroEL and proteases such as the proteasome, comprise protein rings that are stacked on top of one another. While there is some experimental data regarding how stacked-ring complexes such as the proteasome self-assemble, a comprehensive understanding of the dynamics of stacked-ring assembly is currently lacking. Here, we developed a mathematical model of stacked-trimer assembly and performed an analysis of the assembly of the stacked homomeric trimer, which is the simplest stacked-ring architecture. We found that stacked rings are particularly susceptible to a form of kinetic trapping that we term “deadlock,” in which the system gets stuck in a state where there are many large intermediates that are not the fully assembled structure but that cannot productively react. When interaction affinities are uniformly strong, deadlock severely limits assembly yield. We thus predicted that stacked rings would avoid situations where all interfaces in the structure have high affinity. Analysis of available crystal structures indicated that indeed the majority—if not all—of stacked trimers do not contain uniformly strong interactions. Finally, to better understand the origins of deadlock, we developed a formal pathway analysis and showed that, when all the binding affinities are strong, many of the possible pathways are utilized. In contrast, optimal assembly strategies utilize only a small number of pathways. Our work suggests that deadlock is a critical factor influencing the evolution of macromolecular machines and provides general principles for understanding the self-assembly efficiency of existing machines.

DNA Self-Assembly Optimization by Betaine and Its Analogs.

The self-assembly yield of DNA nanostructures can be exponentially lower with increasing structural complexity. Few optimizing strategies are available in the DNA nanotechnology field for the assembly yield improvement. Here, betaine and its analogs are applied as supplementary ingredients in DNA self-assembly. Such a simple implementation results in effective yield improvement. Through a comprehensive investigation, a reliable yield improvement of two- to threefold is achieved for a number of DNA nanostructures with considerable complexity.

Heteromultivalent DNA Enhances the Assembly Yield of Hybrid Nanoparticles and Facilitates Dynamic Disassembly for Bioanalysis Using ICP-MS.

To obtain enhanced physical and biological properties, various nanoparticles are typically assembled into hybrid nanoparticles through the binding of multiple homologous DNA strands to their complementary counterparts, commonly referred to as homomultivalent assembly. However, the poor binding affinity and limited controllability of homomultivalent disassembly restrict the assembly yield and dynamic functionality of the hybrid nanoparticles. To achieve a higher binding affinity and flexible assembly choice, we utilized the paired heteromultivalency DNA to construct hybrid nanoparticles and demonstrate their excellent assembly characteristics and dynamic applications. Specifically, through heteromultivalency, DNA-functionalized magnetic beads (MBs) and gold nanoparticles (AuNPs) were efficiently assembled. By utilizing ICP-MS, the assembly efficiency of AuNPs on MBs was directly monitored, enabling quantitative analysis and optimization of heteromultivalent binding events. As a result, the enhanced assembly yield is primarily attributed to the fact that heteromultivalency allows for the maximization of effective DNA probes on the surface of nanoparticles, eliminating steric hindrance interference. Subsequently, with external oligonucleotides as triggers, it was revealed that the disassembly mechanism of hybrid nanoparticles was initiated, which was based on an increased local concentration rather than toehold-mediated displacement of paired heteromultivalency DNA probes. Capitalizing on these features, an output platform was then established based on ICP-MS signals that several Boolean operations and analytical applications can be achieved by simply modifying the design sequences. The findings provide new insights into DNA biointerface interaction, with potential applications to complex logic operations and the construction of large DNA nanostructures.

Barium Concentration-Dependent Anomalous Electrophoresis of Synthetic DNA Motifs.

The structural integrity, assembly yield, and biostability of DNA nanostructures are influenced by the metal ions used to construct them. Although high (>10 mM) concentrations of divalent ions are often preferred for assembling DNA nanostructures, the range of ion concentrations and the composition of the assembly products vary for different assembly conditions. Here, we examined the unique ability of Ba2+ to retard double crossover DNA motifs by forming a low mobility species, whose mobility on the gel is determined by the concentration ratio of DNA and Ba2+. The formation of this electrophoretically retarded species is promoted by divalent ions such as Mg2+, Ca2+, and Sr2+ when combined with Ba2+ but not on their own, while monovalent ions such as Na+, K+, and Li+ do not have any effect on this phenomenon. Our results highlight the complex interplay between the metal ions and DNA self-assembly and could inform the design of DNA nanostructures for applications that expose them to multiple ions at high concentrations.

Predisposition in Dynamic Covalent Chemistry: The Role of Non-Covalent Interactions in the Assembly of Tetrahedral Boronate Cages.

Directional bonding strategies guide the design of complex molecular architectures, yet challenges arise due to emergent behavior. Rigid structures face geometric constraints and sensitivity to mismatches, hindering the efficient assembly of molecular organic cages (MOCs). Harnessing intramolecular non-covalent interactions offers a promising solution, broadening geometrical possibilities and enhancing adaptability to boost assembly yields. However, identifying these interactions remains challenging, with their full potential sometimes latent until final assembly. This study explores these challenges by synthesizing boronic acid tripods with varied oxygen positions at the tripodal feet and investigating their role in assembling tetrahedral boronate MOCs. Our results reveal substantial differences in the assembly efficiency among tripods. While the building blocks with oxygen in the benzylic position relative to the central aromatic ring form the MOCs in high yields, those with the oxygen atom directly bound to the central aromatic ring, only yield traces. Through X-ray crystallography and DFT analyses, we elucidate how intramolecular interactions profoundly influence the geometry of the building blocks and cages in a relay-like fashion, highlighting the importance of considering intramolecular interactions in the rational design of (supra)molecular architectures.

Microscale underfill dynamics and void formation of high-density flip-chip packaging: Experiments and simulations

This research focuses on developing high-fidelity experimental and numerical models to analyze microscale underfill dynamics and void formation in high-density flip-chip packaging. Three underfilling scenarios are investigated, namely, point type, I-shaped line type, and L-type line type. The point-type underfilling validates the numerical model against experimental results, while the I-shaped and L-type line-type underfilling explore grid independence, void formation, and critical parameters such as filling position, contact angle, and liquid viscosity. Results indicate that contact angle and viscosity significantly influence filling efficiency and interface evolution. A smaller contact angle accelerates the process, reducing interface jumping motions. Viscous effects are quantified, revealing dimensionless filling time convergence. The use of low-viscosity surrogate fluids enhances numerical simulation efficiency. Sub-bump-sized, bump-sized, and sup-bump-sized voids are observed, identifying three void formation scenarios representing different underfilling flow mechanisms. This study provides insight into microscale flip-chip underfill physics and establishes validated models for next-generation high-density flip-chip products. These models can be further refined and integrated into optimization tools for automated process design, contributing to improved assembly yield and reliability of emerging electronic packages through physics-based understanding and modeling.

Seleno-relaxin analogues: effect of internal and external diselenide bonds on the foldability and a fibrosis-related factor of endometriotic stromal cells.

Human relaxin-2 (H2 relaxin) is a peptide hormone of about 6 kDa, first identified as a reproductive hormone involved in vasoregulation during pregnancy. It has recently attracted strong interest because of its diverse functions, including anti-inflammatory, anti-fibrotic, and vasodilatory, and has been suggested as a potential peptide-based drug candidate for a variety of diseases. Mature H2 relaxin is constituted by the A- and B-chains stabilized by two interchain disulfide (SS) bridges and one intrachain SS linkage. In this study, seleno-relaxins, SeRlx-α and SeRlx-β, which are [C11UA,C11UB] and [C10UA,C15UA] variants of H2 relaxin, respectively, were synthesized via a one-pot oxidative chain assembly (folding) from the component A- and B-chains. The substitution of SS bonds in a protein with their analogue, diselenide (SeSe) bonds, has been shown to alter the physical, chemical, and physiological properties of the protein. The surface SeSe bond (U11A-U11B) enhanced the yield of chain assembly while the internal SeSe bond (U10A-U15A) improved the reaction rate of the folding, indicating that these bridges play a major role in controlling the thermodynamics and kinetics, respectively, of the folding mechanism. Furthermore, SeRlx-α and SeRlx-β effectively reduced the expression of a tissue fibrosis-related factor in human endometriotic stromal cells. Thus, the findings of this study indicate that the S-to-Se substitution strategy not only enhances the foldability of relaxin, but also provides new guidance for the development of novel relaxin formulations for endometriosis treatment.

A computational toolbox for the assembly yield of complex and heterogeneous structures

The self-assembly of complex structures from a set of non-identical building blocks is a hallmark of soft matter and biological systems, including protein complexes, colloidal clusters, and DNA-based assemblies. Predicting the dependence of the equilibrium assembly yield on the concentrations and interaction energies of building blocks is highly challenging, owing to the difficulty of computing the entropic contributions to the free energy of the many structures that compete with the ground state configuration. While these calculations yield well known results for spherically symmetric building blocks, they do not hold when the building blocks have internal rotational degrees of freedom. Here we present an approach for solving this problem that works with arbitrary building blocks, including proteins with known structure and complex colloidal building blocks. Our algorithm combines classical statistical mechanics with recently developed computational tools for automatic differentiation. Automatic differentiation allows efficient evaluation of equilibrium averages over configurations that would otherwise be intractable. We demonstrate the validity of our framework by comparison to molecular dynamics simulations of simple examples, and apply it to calculate the yield curves for known protein complexes and for the assembly of colloidal shells.

Hybrid Bond Interconnect and Advanced Packaging Solutions

Hybrid bond interconnect is a scalable technology that offers significant improvements over the incumbent Cu mbump technology. The notable advantages include, enhanced reliability of elemental metal interconnect, improved thermal performance of an inorganic bond interface and the high-speed performance with a low capacitance and inductance pad structure. Currently, the industry is adopting this technology for high-performance compute applications; however, we anticipate a broader adoption once the technology is readily available within the full supply chain. In this paper, we present the performance of our hybrid bond interconnect on multiple test vehicles including single 1 to 8-die stacks with various pad-pitch sizes, die sizes and configurations. We share die to wafer assembly yield results using our manufacturing worthy process where die are prepared on tape frame using standard pick and place equipment. The test vehicles cover a range of pad sizes (2-15 um), with pitches (4-40 um) and have different x,y dimensions from 1mmx1mm to 9mmx12mm. Reliability tests include MSL-3 precondition, high temperature storage and thermal cycling and on the various test structures

Ratiometric Fluorescence Biosensing of Tandem Biemissive Ag Clusters Boosted by Confined Catalytic DNA Assembly.

The reaction kinetics and yield of traditional DNA assembly with a low local concentration in homogeneous solution remain challenging. Exploring confined catalytic DNA assembly (CCDA) is intriguing to boost the reaction rate and efficacy for creating rapid and sensitive biosensing platforms. A rolling circle amplification (RCA) product containing multiple tandem repeats is a natural scaffold capable of guiding the periodic assembly of customized functional probes at precise sites. Here, we present a RCA-confined CCDA strategy to speed up amplifiable conversion for ratiometric fluorescent sensing of a sequence-specific inducer (I*) by using string green-/red-Ag clusters (sgAgCs and srAgCs) as two counterbalance emitters. Upon recognition of I*, CCDA events are operated by two toehold-mediated strand displacements and localized in repetitive units, thereby releasing I* for recycled signal amplification in the as-grown RCA concatemer. The local concentration of reactive species is increased to facilitate rapider dsDNA complex assembly and more efficient input-output conversion, on which the clustering template sequences of sgAgCs and srAgCs are blocked and opened, enabling srAgCs synthesis but opposite to sgAgCs. Thus, the fluorescence emission of srAgCs goes up, while sgAgCs go down. With the resultant ratio featuring inherent built-in correction, rapid, sensitive, and accurate quantification of I* at the picomolar level is achieved. Benefiting from efficient RCA confinement to enhance reaction kinetics and conversion yield, this CCDA-based strategy provides a new paradigm for developing simple and diverse biosensing methodologies.

Assembly Solutions for Cost-Effective Heterogeneous Integration with Disparate Die Types

The semiconductor industry is driving to enable high volume integration of disparate die types via Heterogeneous Integration. These die can come from a range of wafer sizes fabricated in different technology nodes. This emerging package type creates new challenges regarding assembly efficiency and yield. Traditionally, flip-chip assembly process flows have utilized a single placement tool for the placement of the single die type onto the target substrate. For applications with multiple die types, a series of placement tools have been configured in a production line, with each tool dedicated to the placement of a specific die type. This paper and presentation explore the implications of this type of solution in the era of Heterogeneous Integration. Impacts on product yield, throughput, manufacturing efficiency, and overall cost of assembly will be explored for a broad range of Heterogeneous Integration die configurations. Based on a sensitivity analysis for the range of die types expected in these applications, a novel approach to optimization of overall assembly economics will be proposed. Appropriateness of this novel approach will be explored for a range of packaging solutions, including Flip Chip, 2.5D, 3D, and Fan-Out.

Influence of Rigid Carrier Substrates & its Release Layer on Ultra Fine Pitch chip last FO-WLPs

In recent years the leading-edge product enablers in Wireless Connectivity for WiFi & Bluetooth, Wearable HD Modules, Artificial Intelligence (AI), Application Processors etc. require ultra-high density chip packaging device using heterogenous integration to constructing the SiP modules. In order to meet their requirements, fan-out chip last WLP – having its redistribution layer (RDL) constructed first – is definitely the solution of choice to assemble devices in such advanced packaging platforms. This approach further eliminates known good die (KGD) loss risk, resulting in higher assembly yield and lower cost. Rigid carrier substrate and the release layer placed on top of it influences the process yield of fan-out chip last WLP. It gets more challenging when ultra-fine pitch redistribution layers (RDL) are required. This paper focuses on the influence of release layer on quality of narrow line & space RDL, as well as on the separation force during debonding. It further describes an optimized solution through HRDP® (High Resolution Debondable Panel) technology, which is the most compatible carrier to fabricate the leading-edge fan-out chip last process. The HRDP® carrier is offered in various dimensions and thicknesses as round wafers and square/rectangular panels with glass or silicon to match customer requirements. This is made of 100% inorganic material, which helps to improve interfacial stresses and facilitates debonding. During processing, this inorganic release layer (IRL) demonstrates higher chemical resistance and heat resistance than alternative technology such as the laser lift-off (LLO) type carrier.

Functional characterization of modules for the Silicon Tracking System of the CBM experiment

The CBM experiment is a fixed-target heavy-ion physics experiment at the future FAIR facility, designed to explore the QCD phase diagram at high baryon-net densities and moderate temperatures. To address its complex physics program, the experiment is conceived to operate at beam–target interaction rates up to 10 MHz with self-triggered front-end electronics and free-streaming readout architecture. The Silicon Tracking System (STS) of CBM is the main charged-particle tracker, capable of delivering time-stamped hits for online tracking and event building, and momentum measurement inside a superconducting dipole magnet. The functional unit of the STS is the detector module, a device assembled from a double-sided silicon microstrip sensor connected via a stack of microcables to a pair of front-end boards carrying the readout electronics. The assembly and successful operation of STS modules, produced with close-to-final components, was an important milestone achieved in recent years. The systematic study of the modules’ performance is also necessary for optimizing the assembly procedure and ensuring high production yield. This work reports a series of tests, which aim to evaluate the module’s operation and establish the basic criteria for quality control. It summarizes the procedures and results of different measurements performed with the modules, starting with noise estimation and charge amplitude calibration, resulting in the linearization of the channel’s ADC and a homogeneous threshold distribution. Those results were verified using a radioactive source. Additionally, first conclusions concerning the assembly yield in terms of number of functional channels were obtained. These studies represent a fundamental step towards the series production readiness for the final STS detector.

Fabrication of hierarchical nanostructures using binary colloidal nanosphere assembly

In this paper, we investigate the self-assembly of hierarchical nanostructures using monodispersed nanospheres with two different diameters. Our approach is to use a two-step method where the assembly of larger 200 nm nanospheres is used to direct the assembly of smaller 50 nm particles. This self-assembly technique is based on Langmuir–Blodgett assembly and has low equipment cost when compared with traditional lithography methods. We examine the effects of substrate surface treatment, solution concentration ratio, and spin speeds on the quality of the hierarchical assembly. The fabricated samples are examined using optical and scanning electron microscopy to investigate assembly yield. Various defect types are identified and mitigated by process control. The ability to create more complex assembly can result in smaller features and can enhance the performance of photonics and nanostructured surfaces.

Pushing the Limits of Capillary Assembly for the Arbitrary Positioning of Sub-50nm Nanocubes in Printable Plasmonic Surfaces.

The fabrication of high quality nanophotonic surfaces for integration in optoelectronic devices remains a challenge because of the complexity and cost of top-down nanofabrication strategies. Combining colloidal synthesis with templated self-assembly emerged as an appealing low-cost solution. However, it still faces several obstacles before integration in devices can become a reality. This is mostly due to the difficulty in assembling small nanoparticles (<50 nm) in complex nanopatterns with a high yield. In this study, a reliable methodology is proposed to fabricate printable nanopatterns with an aspect ratio varying from 1 to 10 and a lateral resolution of 30 nm via nanocube assembly and epitaxy. Investigating templated assembly via capillary forces, a new regime was identified that was used to assemble 30-40 nm nanocubes in a patterned polydimethylsiloxane template with a high yield for both Au and Ag with multiple particles per trap. This new method relies on the generation and control of an accumulation zone at the contact line that is thin as opposed to dense, displaying higher versatility. This is in contrast with conventional wisdom, identifying a dense accumulation zone as a requirement for high-yield assembly. In addition, different formulations are proposed that can be used for the colloidal dispersion, showing that the standard water-surfactant solutions can be replaced by surfactant-free ethanol solutions, with good assembly yield. This allows to minimize the presence of surfactants that can affect electronic properties. Finally, it is shown that the obtained nanocube arrays can be transformed into continuous monocrystalline nanopatterns via nanocube epitaxy at near ambient temperature, and transferred to different substrates via contact printing. This approach opens new doors to the templated assembly of small colloids and could find potential applications in various optoelectronic devices ranging from solar cells to light-emitting diodes anddisplays.

Self-Assembly of DNA Nanostructures in Different Cations.

The programmable nature of DNA allows the construction of custom-designed static and dynamic nanostructures, and assembly conditions typically require high concentrations of magnesium ions that restricts their applications. In other solution conditions tested for DNA nanostructure assembly, only a limited set of divalent and monovalent ions are used so far (typically Mg2+ and Na+ ). Here, weinvestigate the assembly of DNA nanostructures in a wide variety of ions using nanostructures of different sizes: a double-crossover motif (76 bp), a three-point-star motif (~134 bp), a DNA tetrahedron (534 bp) and a DNA origami triangle (7221 bp). Weshow successful assembly of a majority of these structures in Ca2+ , Ba2+ , Na+ , K+ and Li+ and provide quantified assembly yields using gel electrophoresis and visual confirmation of a DNA origami triangle using atomic force microscopy. Wefurther show that structures assembled in monovalent ions (Na+ , K+ and Li+ ) exhibit up to a 10-fold higher nuclease resistance compared to those assembled in divalent ions (Mg2+ , Ca2+ and Ba2+ ). Ourwork presents new assembly conditions for a wide range of DNA nanostructures with enhanced biostability.

53‐5: Late‐News Paper: High‐resolution Optical Inspection of NR LEDs‐based Assembly and Their Evaluation for Display Applications

This study presents high‐resolution optical inspection methods to evaluate the assembly yield of densely‐aligned NR LED array for self‐emitting inorganic material‐based display. Fast line scanning images and their deep learning‐based image analysis enabled us to quantitively investigate the assembly yield of NR LEDs even up to 5300 pixel per inch in which, thereafter, automatic optical inspection (AOI) accuracy reaches up to 97 % after 8000 times iteration. Furthermore, confocal photoluminescence optical microscopic system with picoseconds time resolving capability investigated tens nanometer spatially‐resolved emission characteristics of NR LEDs while also showing the non‐radiative recombination from the damaged active layers. Therefore, we believe that this study contributes a significant step toward inspection process of micro LEDs and their manufacturing development.

Analysis of Sputtering Yield Measurements for Ion Thruster Grid Materials

Grid assembly is one of the key components of an ion thruster and directly affects the performance and life of the thruster. The measurement of the sputtering yield of the grid assembly under ion beam bombardment is highly significant for predicting the lifetime of the grid assembly and ion thruster. This study systematically summarizes the main methods currently used for sputtering yield measurement of grid materials, analyzes the advantages and disadvantages of different measurement methods, and provides suggestions for sputtering yield measurements in the low-energy () range. In addition, this study compares the sputtering resistance properties of metal- and carbon-based grid materials and summarizes the influence of key core parameters, such as the surface roughness, surface morphology, binding energy, and incident angle, on the sputtering yield. The results can be used to guide the correction of the sputter yield theoretical formula and the numerical simulation of sputtering erosion.

Assembly Solutions for Cost-Effective Heterogeneous Integration with Disparate Die Types

The semiconductor industry is driving to enable high volume integration of disparate die types via Heterogeneous Integration. These die can come from a range of wafer sizes fabricated in different technology nodes. This emerging package type creates new challenges regarding assembly efficiency and yield. Traditionally, flip-chip assembly process flows have utilized a single placement tool for the placement of the single die type onto the target substrate. For applications with multiple die types, a series of placement tools have been configured in a production line, with each tool dedicated to the placement of a specific die type. This paper and presentation explore the implications of this type of solution in the era of Heterogeneous Integration. Impacts on product yield, throughput, manufacturing efficiency, and overall cost of assembly will be explored for a broad range of Heterogeneous Integration die configurations. Based on a sensitivity analysis for the range of die types expected in these applications, a novel approach to optimization of overall assembly economics will be proposed. Appropriateness of this novel approach will be explored for a range of packaging solutions, including Flip-Chip, 2.5D, 3D, and Fan-Out.

Designing the Self-Assembly of Arbitrary Shapes Using Minimal Complexity Building Blocks.

The design space for self-assembled multicomponent objects ranges from a solution in which every building block is unique to one with the minimum number of distinct building blocks that unambiguously define the target structure. We develop a pipeline to explore the design spaces for a set of structures of various sizes and complexities. To understand the implications of the different solutions, we analyze their assembly dynamics using patchy particle simulations and study the influence of the number of distinct building blocks, and the angular and spatial tolerances on their interactions, on the kinetics and yield of the target assembly. We show that the resource-saving solution with a minimum number of distinct blocks can often assemble just as well (or faster) than designs where each building block is unique. We further use our methods to design multifarious structures, where building blocks are shared between different target structures. Finally, we use coarse-grained DNA simulations to investigate the realization of multicomponent shapes using DNA nanostructures as building blocks.