We describe an exciting new application domain for deep reinforcement learning (RL): droplet routing on digital microfluidic biochips (DMFBs). A DMFB consists of a two-dimensional electrode array, and it manipulates droplets of liquid to automatically execute biochemical protocols for clinical chemistry. However, a major problem with DMFBs is that electrodes can degrade over time. The transportation of droplet transportation over these degraded electrodes can fail, thereby adversely impacting the integrity of the bioassay outcome. We demonstrated that the formulation of droplet transportation as an RL problem enables the training of deep neural network policies that can adapt to the underlying health conditions of electrodes and ensure reliable fluidic operations. We describe an RL-based droplet routing solution that can be used for various sizes of DMFBs. We highlight the reliable execution of an epigenetic bioassay with the RL droplet router on a fabricated DMFB. We show that the use of the RL approach on a simple micro-computer (Raspberry Pi 4) leads to acceptable performance for time-critical bioassays. We present a simulation environment based on the OpenAI Gym Interface for RL-guided droplet routing problems on DMFBs. We present results on our study of electrode degradation using fabricated DMFBs. The study supports the degradation model used in the simulator.

The micro-electrode-dot-array (MEDA) architecture provides precise droplet control and real-time sensing in digital microfluidic biochips. Previous work has shown that trapped charge under microelectrodes (MCs) leads to droplets being stuck and failures in fluidic operations. A recent approach utilizes real-time sensing of MC health status, and attempts to avoid degraded electrodes during droplet routing. However, the problem with this solution is that the computational complexity is unacceptable for MEDA biochips of realistic size. Consequently, in this work, we introduce a deep reinforcement learning (DRL)-based approach to bypass degraded electrodes and enhance the reliability of routing. The DRL model utilizes the information of health sensing in real time to proactively reduce the likelihood of charge trapping and avoid using degraded MCs. Simulation results show that our approach provides effective routing strategies for COVID-19 testing protocols. We also validate our DRL-based approach using fabricated prototype biochips. Experimental results show that the developed DRL model completed the routing tasks using a fewer number of clock cycles and shorter total execution time, compared with a baseline routing method. Moreover, our DRL-based approach provides reliable routing strategies even in the presence of degraded electrodes. Our experimental results show that the proposed DRL-based routing is robust to occurrences of electrode faults, as well as increases the lifetime and usability of microfluidic biochips compared to existing strategies.

Digital microfluidic biochips (DMFBs) based on a micro-electrode-dot-array (MEDA) architecture provide fine-grained control and sensing of droplets in real-time. However, excessive actuation of microelectrodes in MEDA biochips can lead to charge trapping during bioassay execution, causing the failure of microelectrodes and erroneous bioassay outcomes. A recently proposed enhancement to MEDA allows run-time measurement of microelectrode health information, thereby enabling synthesis of adaptive routing strategies for droplets. However, existing synthesis solutions are computationally infeasible for large MEDA biochips that have been commercialized. In this paper, we propose a synthesis framework for adaptive droplet routing in MEDA biochips via deep reinforcement learning (DRL). The framework utilizes the real-time microelectrode health feedback to synthesize droplet routes that proactively minimize the likelihood of charge trapping. We show how the adaptive routing strategies can be synthesized using DRL. We implement the DRL agent, the MEDA simulation environment, and the bioassay scheduler using the OpenAI Gym environment. Our framework obtains adaptive routing policies efficiently for COVID-19 testing protocols on large arrays that reflect the sizes of commercial MEDA biochips available in the marketplace, significantly increasing probabilities of successful bioassay completion compared to existing methods.

Silent Data Corruption (SDC) is one of the critical problems in the field of testing, where errors or corruption do not manifest externally. As a result, there is increased focus on improving the outgoing quality of dies by striving for better correlation between structural and functional patterns to achieve a low DPPM. This is very important for NVIDIA's chips due to the various markets we target; for example, automotive and data center markets have stringent in-field testing requirements. One aspect of these efforts is to also target better testability while incurring lower test cost. Since structural testing is faster than functional tests, it is important to make these structural test patterns as effective as possible and free of test escapes. However, with the rising cell count in today's digital circuits, it is becoming increasingly difficult to sensitize faults and propagate the fault effects to scan-flops or primary outputs. Hence, methods to insert observation points to facilitate the detection of hard-to-detect (HtD) faults are being increasingly explored. In this work, we propose an Observation Point Insertion (OPI) scheme using deep learning with the motivation of achieving - 1) better quality test points than commercial EDA tools leading to a potential lower pattern count 2) faster turnaround time to generate the test points. In order to achieve better pattern compaction than commercial EDA tools, we employ Graph Convolutional Networks (GCNs) to learn the topology of logic circuits along with the features that influence its testability. The graph structures are subsequently used to train two GCN-type deep learning models - the first model predicts signal probabilities at different nets and the second model uses these signal probabilities along with other features to predict the reduction in test-pattern count when OPs are inserted at different locations in the design. The features we consider include structural features like gate type, gate logic, reconvergent-fanouts and testability features like SCOAP. Our simulation results indicate that the proposed machine learning models can predict the probabilistic testability metrics with reasonable accuracy and can identify observation points that reduce pattern count.

A digital microfluidic biochip (DMFB) enables the miniaturization of immunoassays, point-of-care clinical diagnostics, and DNA sequencing. A recent generation of DMFBs uses a microelectrode-dot-array (MEDA) architecture, which provides fine-grained control of droplets and real-time droplet sensing using CMOS technology. However, microelectrodes in a MEDA biochip can degrade due to charge trapping when they are repeatedly charged and discharged during bioassay execution; such degradation leads to the failure of microelectrodes and erroneous bioassay outcomes. To address this problem, we first introduce a new microelectrode-cell design such that we can obtain the health status of all the microelectrodes in a MEDA biochip by employing the inherent sensing mechanism. Next, we present a stochastic game-based model for droplet manipulation, and a formal synthesis method for droplet routing that can dynamically change droplet transportation routes. This adaptation is based on the real-time health information obtained from microelectrodes. Comprehensive simulation results for four real-life bioassays show that our method increases the likelihood of successful bioassay completion with negligible impact on time-to-results.

A digital microfluidic biochip (DMFB) enables the miniaturization of immunoassays, point-of-care clinical diagnostics, and DNA sequencing. A recent generation of DMFBs uses a micro-electrode-dot-array (MEDA) architecture, which provides fine-grained control of droplets and real-time droplet sensing using CMOS technology. However, microelectrodes in a MEDA biochip can degrade due to charge trapping when they are repeatedly charged and discharged during bioassay execution; such degradation leads to the failure of microelectrodes and erroneous bioassay outcomes. To address this problem, we first introduce a new microelectrode-cell design such that we can obtain the health status of all the microelectrodes in a MEDA biochip by employing the inherent sensing mechanism. Next, we present a stochastic game-based model for droplet manipulation, and a formal synthesis method for droplet routing that can dynamically change droplet transportation routes. This adaptation is based on the real-time health information obtained from microelectrodes. Comprehensive simulation results for four real-life bioassays show that our method increases the likelihood of successful bioassay completion with negligible impact on time-to-results.

Microfluidic biochips are being utilized for clinical diagnostics, including COVID-19 testing, because of they provide sample-to-result turnaround at low cost. Recently, microelectrode-dot-array (MEDA) biochips have been proposed to advance microfluidics technology. A MEDA biochip manipulates droplets of nano/picoliter volumes to automatically execute biochemical protocols. During bioassay execution, droplets are transported in parallel to achieve high-throughput outcomes. However, a major concern associated with the use of MEDA biochips is microelectrode degradation over time. Recent work has shown that formulating droplet transportation as a reinforcement-learning (RL) problem enables the training of policies to capture the underlying health conditions of microelectrodes and ensure reliable fluidic operations. However, the above RL-based approach suffers from two key limitations: 1) it cannot be used for concurrent transportation of multiple droplets; 2) it requires the availability of CCD cameras for monitoring droplet movement. To overcome these problems, we present a multi-agent reinforcement learning (MARL) droplet-routing solution that can be used for various sizes of MEDA biochips with integrated sensors, and we demonstrate the reliable execution of a serial-dilution bioassay with the MARL droplet router on a fabricated MEDA biochip. To facilitate further research, we also present a simulation environment based on the PettingZoo Gym Interface for MARL-guided droplet-routing problems on MEDA biochips.

A digital microfluidic biochip (DMFB) enables the miniaturization of immunoassays, point-of-care clinical diagnostics, DNA sequencing, and other laboratory procedures in biochemistry. A recent generation of biochips uses a micro-electrode-dot-array (MEDA) architecture, which provides fine-grained control of droplets and seamlessly integrates microelectronics and microfluidics using CMOS technology and a TSMC fabrication process. To ensure that bioassays are carried out on MEDA biochips efficiently, high-level synthesis algorithms have recently been proposed. However, as in the case of conventional DMFBs, microelectrodes are likely to fail when they are heavily utilized, and previous methods fail to consider reliability issues. In this article, we first present a new microelectrode cell (MC) design such that the droplet-sensing operation can be enabled/disabled for individual MCs. Next, “partial update” and “partial sensing” operations are presented based on an IEEE Std. 1687 IJTAG network design. Finally, wear-leveling synthesis method is proposed to ensure uniform utilization of MCs on MEDA. A comprehensive set of simulation results demonstrate the effectiveness of the proposed hardware design and design automation methods.

A digital microfluidic biochip (DMFB) enables miniaturization of immunoassays, point-of-care clinical diagnostics, DNA sequencing, and other laboratory procedures in biochemistry. A recent generation of biochips uses a microelectrode-dot-array (MEDA) architecture, which provides fine-grained control of droplets and seamlessly integrates microelectronics and microfluidics using CMOS technology. To ensure that bioassays are carried out on MEDA biochips efficiently, high-level synthesis algorithms have recently been proposed. However, as in the case of conventional DMFBs, microelectrodes are likely to fail when they are heavily utilized, and previous methods fail to consider reliability issues. In this paper, we present the design of an IEEE Std. 1687 (IJTAG) network and a block-aware high-level synthesis method that can effectively alleviate reliability problems in MEDA biochips. A comprehensive set of simulation results demonstrate the effectiveness of the proposed method.

We present and investigate a novel application domain for deep reinforcement learning (RL): droplet routing on digital microfluidic biochips (DMFBs). A DMFB, composed of a two-dimensional electrode array, manipulates discrete fluid droplets to automatically execute biochemical protocols such as point-of-care clinical diagnosis. However, a major concern associated with the use of DMFBs is that electrodes in a biochip can degrade over time. Droplet-transportation operations associated with the degraded electrodes can fail, thereby compromising the integrity of the bioassay outcome. We show that casting droplet transportation as an RL problem enables the training of deep network policies to capture the underlying health conditions of electrodes and to provide reliable fluidic operations. We propose a new RL-based droplet-routing flow that can be used for various sizes of DMFBs, and demonstrate reliable execution of an epigenetic bioassay with the RL droplet router on a fabricated DMFB. To facilitate further research, we also present a simulation environment based on the OpenAI Gym Interface for RL-guided droplet-routing problems on DMFBs.

Sample preparation, as a key procedure in many biochemical protocols, mixes various samples, and/or reagents into solutions that contain the target concentrations. Digital microfluidic biochips (DMFBs) have been adopted as a platform for sample preparation because they provide automatic procedures that require less reactant consumption and reduce human-induced errors. However, the most existing methods only consider two-reactant sample preparation, and they cannot be used for many biochemical applications that involve multiple reactants. In addition, the existing methods that can be used for multiple-reactant sample preparation were proposed on traditional DMFBs where only the (1:1) mixing model is available. In the (1:1) mixing model, only two droplets of the same volume can be mixed at a time, which results in higher completion time and the wastage of valuable reactants. To overcome this limitation, the micro-electrode-dot-array (MEDA) architecture has been introduced; it provides the flexibility of mixing multiple droplets of different volumes in a single operation. In this article, we present a generic multiple-reactant sample preparation algorithm that exploits the novel fluidic operations on MEDA biochips. We also propose an enhanced algorithm that increases the operation-sharing opportunities when multiple target concentrations are needed, and therefore the usage of reactants can be further reduced. The simulated experiments show that the proposed method outperforms existing methods in terms of saving reactant cost, minimizing the number of operations, and reducing the amount of waste.

A digital microfluidic biochip (DMFB) enables miniaturization of immunoassays, point-of-care clinical diagnostics, and DNA sequencing. A recent generation of DMFBs uses a micro-electrode-dot-array (MEDA) architecture, which provides fine-grained control of droplets and real-time droplet sensing using the CMOS technology. However, microelectrodes in a MEDA biochip degrade when they are charged and discharged frequently during bioassay execution. In this article, we first make the key observation that the droplet-sensing operations contribute up to 94% of all microelectrode actuation in MEDA. Consequently, to reduce the number of droplet-sensing operations, we present a new microelectrode cell (MC) design as well as a selective-sensing method such that only a small fraction of microelectrodes perform droplet sensing during bioassay execution. The selection of microelectrodes that need to perform the droplet sensing is based on an analysis of experimental data. A comprehensive set of simulation results show that the total number of droplet-sensing operations is reduced to only 0.7%, which prolongs the lifespan of a MEDA biochip by 11× without any impact on bioassay time-to-response.

Biochemical experiments, such as diagnostics must be precise and trusted, and provide quick time to results. This has been enabled by automated digital microfluidics; however, it also exposes these experiments to security threats. Previous work has shown that the critical challenge in securing digital microfluidic devices is the lack of sensing resources. The micro-electrode-dot-array (MEDA) is a next-generation digital microfluidic biochip platform that supports fine-grained control and real-time sensing of droplet movements. These capabilities permit continuous monitoring and checkpoint (CP)-based validation of assay execution on MEDA. This article presents a class of “shadow attacks” that abuse the timing slack in the assay execution. State-of-the-art CP-based validation techniques cannot expose the shadow operations. We overcome this limitation by introducing extra CPs in the assay execution at time instances when the assay is prone to shadow attacks. We achieve this by identifying the conditions that enable shadow attacks. We use these conditions to minimize the number of CPs required to guarantee the correctness of bioassay implementation. Our simulation results confirm the effectiveness and practicality of the defense.

DNA fingerprinting can offer remarkable benefits, especially for point-of-care diagnostics, information forensics, and analysis. However, the pressure to drive down costs is likely to lead to cheap untrusted solutions and a multitude of unprecedented risks. These risks will especially emerge at the frontier between the cyberspace and DNA biology. To address these risks, we perform a forensic-security assessment of a typical DNA-fingerprinting flow. We demonstrate, for the first time, benchtop analysis of biochemical-level vulnerabilities in flows that are based on a standard quantification assay known as polymerase chain reaction (PCR). After identifying potential vulnerabilities, we realize attacks using benchtop techniques to demonstrate their catastrophic impact on the outcome of the DNA fingerprinting. We also propose a countermeasure, in which DNA samples are each uniquely barcoded (using synthesized DNA molecules) in advance of PCR analysis, thus demonstrating the feasibility of our approach using benchtop techniques. We discuss how molecular barcoding could be utilized within a cyber-biological framework to improve DNA-fingerprinting security against a wide range of threats, including sample forgery. We also present a security analysis of the DNA barcoding mechanism from a molecular biology perspective.

Digital microfluidic biochips (DMFBs) have emerged as a promising platform for DNA sequencing, clinical chemistry, and point-of-care diagnostics. Recent research has shown that DMFBs are susceptible to various types of malicious attacks. Defenses proposed thus far only offer probabilistic guarantees of security due to the limitation of on-chip sensor resources. A micro-electrode-dot-array (MEDA) biochip is a next-generation DMFB that enables the real-time sensing of on-chip droplet locations, which are captured in the form of a droplet-location map. We propose a security mechanism that validates assay execution by reconstructing the sequencing graph (i.e., the assay specification) from the droplet-location maps and comparing it against the golden sequencing graph. We prove that there is a unique (one-to-one) mapping from the set of droplet-location maps (over the duration of the assay) to the set of possible sequencing graphs. Any deviation in the droplet-location maps due to an attack is detected by this countermeasure because the resulting derived sequencing graph is not isomorphic to the original sequencing graph. We highlight the strength of the security mechanism by simulating attacks on real-life bioassays. We also address the concern that the proposed mechanism may raise false alarms when some fluidic operations are executed on MEDA biochips. To avoid such false alarms, we propose an enhanced sensing technique that provides fine-grained sensing for the security mechanism.

As digital microfluidic biochips (DMFBs) make the transition to the marketplace for commercial exploitation, security and intellectual property (IP) protection are emerging as important design considerations. Recent studies have shown that DMFBs are vulnerable to reverse engineering aimed at stealing biomolecular protocols (IP theft). The IP piracy of proprietary protocols may lead to significant losses for pharmaceutical and biotech companies. The microelectrode dot array (MEDA) is a next-generation DMFB platform that supports real-time sensing of droplets and has the added advantage of important security protection. However, real-time sensing offers opportunities to an attacker to steal the biochemical IP. We show that the daisychaining of microelectrodes and the use of one-time programmability in MEDA biochips provides effective bitstream scrambling of biochemical protocols. To examine the strength of this solution, we develop a Satisfiability (SAT)-based attack that can unscramble the bitstreams through repeated observations of bioassays executed on the MEDA platform. Based on insights gained from the SAT attack, we propose an advanced defense against IP theft. Simulation results using real-life biomolecular protocols confirm that while the SAT attack is effective for simple instances, our advanced defense can thwart it for realistic MEDA biochips and real-life protocols.

Sample preparation, as a key procedure in many biochemical protocols, mixes various samples and/or reagents into solutions that contain the target concentrations. Digital microfluidic biochips (DMFBs) have been adopted as a platform for sample preparation because they provide automatic procedures that require less reactant consumption and reduce human-induced errors. However, traditional DMFBs only utilize the (1:1) mixing model, i.e., only two droplets of the same volume can be mixed at a time, which results in higher completion time and the wastage of valuable reactants. To overcome this limitation, a next-generation micro-electrode-dot-array (MEDA) architecture that provides flexibility of mixing multiple droplets of different volumes in a single operation was proposed. In this paper, we present a generic multiple-reactant sample preparation algorithm that exploits the novel fluidic operations on MEDA biochips. Simulated experiments show that the proposed method outperforms existing methods in terms of saving reactant cost, minimizing the number of operations, and reducing the amount of waste.

Digital microfluidic biochips (DMFBs) have emerged as a promising platform for DNA sequencing, clinical chemistry, and point-of-care diagnostics. Recent research has shown that DMFBs are susceptible to various types of malicious attacks. Defenses proposed thus far only offer probabilistic guarantees of security due to the limitation of on-chip sensor resources. A micro-electrode-dot-array (MEDA) biochip is a next-generation DMFB that enables the sensing of on-chip droplet locations, which are captured in the form of a droplet-location map. We propose a security mechanism that validates assay execution by reconstructing the sequencing graph (i.e., the assay specification) from the droplet-location maps and comparing it against the golden sequencing graph. We prove that there is a unique (one-to-one) mapping from the set of droplet-location maps (over the duration of the assay) to the set of possible sequencing graphs. Any deviation in the droplet-location maps due to an attack is detected by this countermeasure because the resulting derived sequencing graph is not isomorphic to the original sequencing graph. We highlight the strength of the security mechanism by simulating attacks on real-life bioassays.

As digital microfluidic biochips (DMFBs) make the transition to the marketplace for commercial exploitation, security and intellectual property (IP) protection are emerging as important design considerations. Recent studies have shown that DMFBs are vulnerable to reverse engineering aimed at stealing biomolecular protocols (IP theft). The IP piracy of proprietary protocols may lead to significant losses for pharmaceutical and biotech companies. The micro-electrode-dot-array (MEDA) is a next-generation DMFB platform that supports real-time sensing of droplets and has the added advantage of important security protections. However, real-time sensing offers opportunities to an attacker to steal the biochemical IP. We show that the daisychaining of microelectrodes and the use of one-time-programmability in MEDA biochips provides effective bitstream scrambling of biochemical protocols. To examine the strength of this solution, we develop a SAT attack that can unscramble the bitstreams through repeated observations of bioassays executed on the MEDA platform. Based on insights gained from the SAT attack, we propose an advanced defense against IP theft. Simulation results using real-life biomolecular protocols confirm that while the SAT attack is effective for simple instances, our advanced defense can thwart it for realistic MEDA biochips and real-life protocols.

The Micro-electrode-dot-array (MEDA) is a next-generation digital microfluidic biochip (DMFB) platform that supports fine-grained control and real-time sensing of droplet movements. These capabilities permit continuous monitoring and checkpoint-based validation of assay execution on MEDA. This paper presents a class of “shadow attacks” that abuse the timing slack in the assay execution. State-of-the-art checkpoint-based validation techniques cannot expose the shadow operations. We develop a defense that introduces extra checkpoints in the assay execution at time instances when the assay is prone to shadow attacks. Experiments confirm the effectiveness and practicality of the defense.

Sample preparation is an essential processing step in most biochemical applications. Various reactants are mixed together to produce a solution with the target concentration. Since reactants generally take a notable part of the cost in a bioassay, their usage should be minimized whenever possible. In this paper, we propose an algorithm, CoDOS, to prepare the target solution with many reactants using common dilution operation sharing on digital microfluidic biochips (DMFBs). CoDOS first represents the given target concentration as a recipe matrix, and then identifies rectangles in the matrix, where each rectangle indicates an opportunity of dilution operation sharing for reactant minimization. Experimental results demonstrate that CoDOS can achieve up to 27% of reactant saving as compared with the bit-scanning method in single-target sample preparation. Moreover, even if CoDOS is not developed for multi-target sample preparation, it still outperforms the recent state-of-the-art algorithm, RSMA. Hence, it is convincing that CoDOS is a better alternative for many-reactant sample preparation.