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Droplet microfluidics for synthetic biology

Here, synthetic biology is an interdisciplinary field that aims to engineer biological systems for useful purposes. Organism engineering often requires the optimization of individual genes and/or entire biological pathways (consisting of multiple genes). Advances in DNA sequencing and synthesis have recently begun to enable the possibility of evaluating thousands of gene variants and hundreds of thousands of gene combinations. However, such large-scale optimization experiments remain cost-prohibitive to researchers following traditional molecular biology practices, which are frequently labor-intensive and suffer from poor reproducibility. Liquid handling robotics may reduce labor and improve reproducibility, but are themselves expensive and thus inaccessible to most researchers. Microfluidic platforms offer a lower entry price point alternative to robotics, and maintain high throughput and reproducibility while further reducing operating costs through diminished reagent volume requirements. Droplet microfluidics have shown exceptional promise for synthetic biology experiments, including DNA assembly, transformation/transfection, culturing, cell sorting, phenotypic assays, artificial cells and genetic circuits.

59 BASIC BIOLOGICAL SCIENCES↗

Autonomous organic synthesis for redox flow batteries via flexible batch Bayesian optimization

Traditional trial-and-error methods for materials discovery are inefficient to meet the urgent demands posed by the rapid progression of climate change. This urgency has driven the increasing interest in integrating robotics and machine learning into materials research to accelerate experimental learning. However, idealized decision-making frameworks to achieve maximum sampling efficiency are not always compatible with high-throughput experimental workflows inside a laboratory. For multi-step chemical processes, differences in hardware capacities can complicate the digital framework by introducing constraints on the maximum number of samples in each step of the experiment, hence causing varying batch sizes in variable selection within the same batch. Therefore, designing flexible sampling algorithms is necessary to accommodate the multi-step synthesis with practical constraints unique to each high-throughput workflow. In this work, we designed and employed three strategies on a high-throughput robotic platform to optimize the sulfonation reaction of redox-active molecules used in flow batteries. Our strategies adapt to the multi-step experimental workflow, where their formulation and heating steps are separate, causing varying batch size requirements. By strategically sampling using clustering and mixed-variable batch Bayesian optimization, we were able to iteratively identify optimal conditions that maximize the yields. Our work presents a flexible approach that allows tailoring the machine learning decision-making to suit the practical constraints in individual high-throughput experimental platforms, followed by performing resource-efficient yield optimization using available open-source Python libraries.

Tamura, Clara [Univ. of Washington, Seattle, WA (U↗

A novel cell weighing method based on the minimum immobilization pressure for biological applications

A novel weighing method for cells with spherical and other regular shapes is proposed in this paper. In this method, the relationship between the cell mass and the minimum aspiration pressure to immobilize the cell (referred to as minimum immobilization pressure) is derived for the first time according to static theory. Based on this relationship, a robotic cell weighing process is established using a traditional micro-injection system. Experimental results on porcine oocytes demonstrate that the proposed method is able to weigh cells at an average speed of 16.3 s/cell and with a success rate of more than 90%. The derived cell mass and density are in accordance with those reported in other published results. The experimental results also demonstrated that this method is able to detect less than 1% variation of the porcine oocyte mass quantitatively. It can be conducted by a pair of traditional micropipettes and a commercial pneumatic micro-injection system, and is expected to perform robotic operation on batch cells. At present, the minimum resolution of the proposed method for measuring the cell mass can be 1.25 × 10{sup −15 }kg. Above advantages make it very appropriate for quantifying the amount of the materials injected into or moved out of the cells in the biological applications, such as nuclear enucleations and embryo microinjections.

71 CLASSICAL AND QUANTUM MECHANICS, GENERAL PHYSIC↗

Photonic methods and technologies for monitoring the ocean and atmosphere

In this paper, we review photonic methods and technologies that are promising for monitoring the ocean and atmosphere and have been implemented mainly in recent years at the Institute of Automation and Control Processes, Far Eastern Branch, Russian Academy of Sciences. We present results of lidar studies that have made it possible to understand key features of ocean – atmosphere interaction processes under continent – ocean transition conditions, which determine specific features of the atmospheric aerosol distribution, small gaseous components of the atmosphere and its optical characteristics. We consider methods and tools for combined optical and laser fluorescence monitoring of the ocean surface. Particular attention is paid to results of research on remote methods and tools for real-time laser-induced and laser fluorescence environmental monitoring of the ocean, including specialised fibre-optic probes and mobile underwater robotic systems. We present results of the development and investigation of highly sensitive, noise-proof fibre-optic hydro- and seismic/acoustic sensors for remote monitoring of the ocean and robotic systems for underwater laser protection of marine vessels, hydraulic structures and oceanographic instruments against the negative impact of biofouling. (invited paper)

46 INSTRUMENTATION RELATED TO NUCLEAR SCIENCE AND ↗

Digital logic for soft devices

Although soft devices (grippers, actuators, and elementary robots) are rapidly becoming an integral part of the broad field of robotics, autonomy for completely soft devices has only begun to be developed. Adaptation of conventional systems of control to soft devices requires hard valves and electronic controls. This paper describes completely soft pneumatic digital logic gates having a physical scale appropriate for use with current (macroscopic) soft actuators. Each digital logic gate utilizes a single bistable valve—the pneumatic equivalent of a Schmitt trigger—which relies on the snap-through instability of a hemispherical membrane to kink internal tubes and operates with binary high/low input and output pressures. Soft, pneumatic NOT, AND, and OR digital logic gates—which generate known pneumatic outputs as a function of one, or multiple, pneumatic inputs—allow fabrication of digital logic circuits for a set–reset latch, two-bit shift register, leading-edge detector, digital-to-analog converter (DAC), and toggle switch. The DAC and toggle switch, in turn, can control and power a soft actuator (demonstrated using a pneu-net gripper). We note ehese macroscale soft digital logic gates are scalable to high volumes of airflow, do not consume power at steady state, and can be reconfigured to achieve multiple functionalities from a single design (including configurations that receive inputs from the environment and from human users). This work represents a step toward a strategy to develop autonomous control—one not involving an electronic interface or hard components—for soft devices.

36 MATERIALS SCIENCE↗

Hanging droplets from liquid surfaces

Natural and man-made robotic systems use the interfacial tension between two fluids to support dense objects on liquid surfaces. Here, we show that coacervate-encased droplets of an aqueous polymer solution can be hung from the surface of a less dense aqueous polymer solution using surface tension. The forces acting on and the shapes of the hanging droplets can be controlled. Sacs with homogeneous and heterogeneous surfaces are hung from the surface and, by capillary forces, form well-ordered arrays. Locomotion and rotation can be achieved by embedding magnetic microparticles within the assemblies. Direct contact of the droplet with air enables in situ manipulation and compartmentalized cascading chemical reactions with selective transport. Applications including functional microreactors, motors, and biomimetic robots are evident.

36 MATERIALS SCIENCE↗

Designing and Utilizing Material Acceleration Platforms: Need for Workforce Development

In the quest to accelerate scientific discovery, the materials science field is rapidly moving toward the implementation of robotics and artificial intelligence driven workflows. Our recent summer school “Future Labs: Robotic Synthesis Coupled with Machine Learning for Energy Materials” provided learning opportunities for students, researchers, and educators in the materials science community. We describe this experience and provide our perspective on which new directions could be pursued to enable the future workforce to acquire cross-disciplinary skills.

Educational policy↗

A comprehensive review of the lipid cubic phase or in meso method for crystallizing membrane and soluble proteins and complexes

A comprehensive and up-to-date review of the lipid cubic phase or in meso method for crystallizing membrane and soluble proteins and complexes is reported. Recent applications of the method for in situ serial crystallography at X-ray free-electron lasers and synchrotrons are described. The lipid cubic phase or in meso method is a robust approach for crystallizing membrane proteins for structure determination. The uptake of the method is such that it is experiencing what can only be described as explosive growth. This timely, comprehensive and up-to-date review introduces the reader to the practice of in meso crystallogenesis, to the associated challenges and to their solutions. A model of how crystallization comes about mechanistically is presented for a more rational approach to crystallization. The possible involvement of the lamellar and inverted hexagonal phases in crystallogenesis and the application of the method to water-soluble, monotopic and lipid-anchored proteins are addressed. How to set up trials manually and automatically with a robot is introduced with reference to open-access online videos that provide a practical guide to all aspects of the method. These range from protein reconstitution to crystal harvesting from the hosting mesophase, which is noted for its viscosity and stickiness. The sponge phase, as an alternative medium in which to perform crystallization, is described. The compatibility of the method with additive lipids, detergents, precipitant-screen components and materials carried along with the protein such as denaturants and reducing agents is considered. The powerful host and additive lipid-screening strategies are described along with how samples that have low protein concentration and cell-free expressed protein can be used. Assaying the protein reconstituted in the bilayer of the cubic phase for function is an important element of quality control and is detailed. Host lipid design for crystallization at low temperatures and for large proteins and complexes is outlined. Experimental phasing by heavy-atom derivatization, soaking or co-crystallization is routine and the approaches that have been implemented to date are described. An overview and a breakdown by family and function of the close to 200 published structures that have been obtained using in meso-grown crystals are given. Recommendations for conducting the screening process to give a more productive outcome are summarized. The fact that the in meso method also works with soluble proteins should not be overlooked. Recent applications of the method for in situ serial crystallography at X-ray free-electron lasers and synchrotrons are described. The review ends with a view to the future and to the bright prospects for the method, which continues to contribute to our understanding of the molecular mechanisms of some of nature’s most valued proteinaceous robots.

75 CONDENSED MATTER PHYSICS, SUPERCONDUCTIVITY AND↗

Importance Sampling Model-Based Diffusion for Trajectory Optimization

Trajectory optimization for robotic systems remains a challenging problem. This is especially true for robotic systems featuring nonlinear dynamics and many degrees of freedom. Data-based or model-free diffusion has recently been popularized in the fields of artificial intelligence and trajectory optimization. Model-Based Diffusion provides a data-free method of trajectory optimization, trained at runtime on a system dynamics model, suitable for high-dimensional models. This paper examines how importance sampling can enhance the performance of Model-Based Diffusion for trajectory optimization. Here, we quantify the benefits of importance sampling across three long horizon planning tasks. These results show as much as a 13x improvement in sample efficiency depending on environment and optimization parameters.

Golembeski, Seth [Georgia Institute of Technology,↗

Electrical Power Conversion of River and Tidal Power Generator

As renewable generation has become less expensive during recent decades, and it becomes more accepted by the global population, the focus on renewable generation has expanded to include new types with promising future applications, such as river and tidal generation. Although the utilization of power electronics and electric machines in industry is phenomenal, the emphasis on system design is different for various sectors of industry. In precision control, robotics, and weaponry, the design emphasis is on accuracy and reliability with less concern for the cost of the final product. In energy generation, the cost of energy is the prime concern; thus, capital expenditures (CAPEX) and operations and maintenance expenditures (OPEX) are the major design objectives. This paper describes the electrical power conversion aspects of river and tidal generation. Although modern power converter control is available to control the generation side, the design was chosen on the bases of minimizing the CAPEX and OPEX; thus, the architecture is simple and modular for ease of replacement and maintenance. The power conversion is simplified by considering a simple diode bridge and a DC-DC power converter to take advantage of abundant and low-cost photovoltaic inverters that have well-proven grid integration characteristics (i.e., the capability to produce energy with good power quality and control real power and voltage on the grid side).

16 TIDAL AND WAVE POWER↗

HLRW management during MR reactor decommissioning in NRC 'Kurchatov Institute'

A program of decommissioning of MR research reactor in the Kurchatov institute started in 2008. The decommissioning work presumed a preliminary stage, which included: removal of spent fuel from near reactor storage; removal of spent fuel assemble of metal liquid loop channel from a core; identification, sorting and disposal of radioactive objects from gateway of the reactor; identification, sorting and disposal of radioactive objects from cells of HLRW storage of the Kurchatov institute for radwaste creating form the decommissioning of MR. All these works were performed by a remote controlled means with use of a remote identification methods of high radioactive objects. A distribution of activity along high radiated objects was measured by a collimated radiometer installed on the robot Brokk-90, a gamma image of the object was registered by gamma-visor. Spectrum of gamma radiation was measured by a gamma locator and semiconductor detector system. For identification of a presence of uranium isotopes in the HLRW a technique, based on the registration of characteristic radiation of U, was developed. For fragmentation of high radiated objects was used a cold cutting technique and dust suppression system was applied for reduction of volume activity of aerosols in air. The management of HLRW was performed by remote controlled robots Brokk-180 and Brokk-330. They executed sorting, cutting and parking of high radiated part of contaminated equipment. The use of these techniques allowed to reduce individual and collective doses of personal performed the decommissioning. The average individual dose of the personnel was 1,9 mSv/year in 2011, and the collective dose is estimated by 0,0605 man x Sv/year. Use of the remote control machines enables reducing the number of working personal (20 men) and doses. X-ray spectrometric methods enable determination of a presence of the U in high radiated objects and special cans and separation of them for further spent fuel inspection. The sorting of radwaste enabled shipping of the LLRW and ILRW to special repositories and keeping of the HLRW for decay in the Kurchatov institute repository. (authors)

12 MANAGEMENT OF RADIOACTIVE AND NON-RADIOACTIVE W↗

Novel techniques and devices for in-situ film coatings of long, small diameter tubes or elliptical and other surface contours

In this study, devices and techniques that can, via physical vapor deposition,coat various surface contours or very long small aperture pipes, are described. Recently, a magnetron mole was developed in order to in-situ coat accelerator tube sections of the Brookhaven National Lab relativistic heavy ion collider that have 7.1 cm diameter with access points that are 500 m apart, for copper coat the accelerator vacuum tube in order to alleviate the problems of unacceptable ohmic heating and of electron clouds. A magnetron with a 50 cm long cathode was designed fabricated and successfully operated to copper coat a whole assembly containing a full-size, stainless steel, cold bore, of the accelerator magnet tubing connected to two types bellows, to which two additional pipes made of accelerator tubing were connected. The magnetron is mounted on a carriage with spring loaded wheels that successfully crossed bellows and adjusted for variations in vacuum tube diameter, while keeping the magnetron centered. Electrical power and cooling water were fed through a cable bundle. The umbilical cabling system, which is enclosed in a flexible braided metal sleeve, is driven by a motorized spool. To increase cathode lifetime, movable magnet package was developed, and thickest possible cathode was made, with a rather challenging target to substrate distance of less than 1.5 cm. Optimized process to ensure excellent adhesion was developed. Coating thickness of 10 μm Cu passed all industrial tests and even exceeded maximum capability of a 12 kg pull test fixture. Room temperature radio frequency (RF) resistivity measurement indicated that 10 μm Cu coated stainless steel accelerator tube has conductivity close to copper tubing. Work is in progress to repeat the RF resistivity measurement at cryogenic temperatures. Over 20 years ago, a device using multi axis robotic manipulators controlling separate robotic assemblies resulted in nine-axes of motion combined with conformal shape of the cathodes that can adapt to various curved surface contours was developed and successfully used for depositing optical coating on aircraft canopies. The techniques can be utilized for in situ coating of elliptical and other surface contour RF cavities and long beam pipes with thick superconducting films. Plans are to incorporate ion assisted deposition in those techniques for attaining dense, adherent and defect free coatings.

43 PARTICLE ACCELERATORS↗

An elastography method based on the scanning contact resonance of a piezoelectric cantilever

Purpose: Most tissues may become significantly stiffer than their normal states when there are lesions inside. The tissue's modulus can then act as an identification parameter for clinic diagnosis of tumors or fibrosis, which leads to elastography. This study introduces a novel elastography method that can be used for modulus imaging of superficial organs. Methods: This method is based on the scanning contact-resonance of a unimorph piezoelectric cantilever. The cantilever vibrates in its bending mode with the tip pressed tightly on the sample. The contact resonance frequency of the cantilever-sample system is tracked at each scanning point, from which the sample's modulus can be derived based on a beam dynamic model and a contact mechanics model. Scanning is performed by a three-dimensional motorized stage and the whole system is controlled by a homemade software program based on LabVIEW. Results: Testing onin vitro beef tissues indicates that the fat and the muscle can be easily distinguished using this system, and the accuracy of the modulus measurement can be comparable with that of nanoindentation. Imaging on homemade gelatin phantoms shows that the depth information of the abnormalities can be qualitatively obtained by varying the pressing force. The detection limit of this elastography method is specially examined both experimentally and numerically. Results show that it can detect the typical lesions in superficial organs with the depth of several centimeters. The lateral resolution of this elastography method/system is better than 0.5 mm, and could be further enhanced by using more scanning points. Conclusions: The proposed elastography system can be regarded as a sensitive palpation robot, which may be very promising in early diagnosis of tumors in superficial organs such as breast and thyroid.

62 RADIOLOGY AND NUCLEAR MEDICINE↗

SU-E-T-228: Liquid Ionisation Chamber Array and MicroDiamond Measurements with the CyberKnife System

Purpose: The aim of this study was to measure the dose profile and output factors with a CyberKnife accelerator using a TM60019 microDiamond detector and a 1000SRS liquid chamber array (both PTW Freiburg, Germany). Methods: An MP3 water phantom (PTW, Freiburg) was positioned along the robotic world coordinate system. The TM60019 detector was adjusted to the center of the according fields and the semiconductor axis was aligned with the beam direction. Profiles at 5cm water depth and SSD = 80 cm were measured along the robotic x axis and y axis for the cylindrical collimators of the CyberKnife (diameter 60, 50, 40, 30, 20, 15, 12.5, 10, 7.5 and 5mm). To determine the output factors the dose profile was measured at 0.1 mm steps around the field center to find the maximum dose value. The liquid chamber array (1000SRS) measurement was performed with the same setup, but with RW3 buildup. Results: The 1000SRS measurements closely conform with the TM60019 profile measurement in all profile regions and for all collimator sizes. The profile measurement is influenced by the almost equal spatial resolution of the TM60019 detector (radius of the sensitive area 1.1mm) and of the 1000SRS liquid chamber array (single chamber width 2.3mm). The measured dose profiles have not been corrected for this limited spatial resolution. Rather we purpose to consider that spatial dose averaging over 2 mm wide regions might be justified in view of patient positioning inaccuracies and of the spaces in tissue participating in the biological radiation responses. Conclusion: The 1000SRS data points conform with the TM60019 profile measurements at all profile regions showing the applicability of liquid ion chamber arrays with the CyberKnife system.

60 APPLIED LIFE SCIENCES↗

SU-E-T-642: Safety Procedures for Error Elimination in Cyberknife Stereotactic Radiosurgery (SRS) and Stereotactic Body Radiation Therapy (SBRT)

Purpose: Cyberknife system is used for providing stereotactic radiosurgery (SRS) and stereotactic body radiation therapy (SBRT) hypofractionation scheme. The whole treatment delivery is based on live imaging of the patient. The minor error made at any stage may bring severe radiation injury to the patient or damage to the system itself. Several safety measures were taken to make the system safer. Methods: The radiation treatment provided thru a 6MV linac attached to Kuka robot (Cyberknife G4, Accuray Inc. Sunnyvale, CA, USA). Several possible errors were identified related to patient alignment, treatment planning, dose delivery and physics quality assurance. During dose delivery, manual and visual checks were introduced to confirm pre and intra-treatment imaging to reduce possible errors. One additional step was introduced to confirm that software tracking-tools had worked correctly with highest possible confidence level. Robotic head move in different orientations over and around the patient body, the rigidity of linac-head cover and other accessories was checked periodically. The vender was alerted when a tiny or bigger piece of equipment needed additional interlocked support. Results: As of our experience treating 525 patients on Cyberknife during the last four years, we saw on and off technical issues. During image acquisition, it was made essential to follow the site-specific imaging protocols. Adequate anatomy was contoured to document the respective doses. Followed by auto-segmentation, manual tweaking was performed on every structure. The calculation box was enclosing the whole image during the final calculation. Every plan was evaluated on slice-by slice basis. To review the whole process, a check list was maintained during the physics 2nd-check. Conclusion: The implementation of manual and visual additional checks introduced along with automated checks for confirmation was found promising in terms of reduction in systematic errors and making the system safer than before.

60 APPLIED LIFE SCIENCES↗

TU-CD-304-01: FEATURED PRESENTATION and BEST IN PHYSICS (THERAPY): Trajectory Modulated Arc Therapy: Development of Novel Arc Delivery Techniques Integrating Dynamic Table Motion for Extended Volume Treatments

Purpose: Integration of coordinated robotic table motion with inversely-planned arc delivery has the potential to resolve table-top delivery limitations of large-field treatments such as Total Body Irradiation (TBI), Total Lymphoid Irradiation (TLI), and Cranial-Spinal Irradiation (CSI). We formulate the foundation for Trajectory Modulated Arc Therapy (TMAT), and using Varian Developer Mode capabilities, experimentally investigate its practical implementation for such techniques. Methods: A MATLAB algorithm was developed for inverse planning optimization of the table motion, MLC positions, and gantry motion under extended-SSD geometry. To maximize the effective field size, delivery trajectories for TMAT TBI were formed with the table rotated at 270° IEC and dropped vertically to 152.5cm SSD. Preliminary testing of algorithm parameters was done through retrospective planning analysis. Robotic delivery was programmed using custom XML scripting on the TrueBeam Developer Mode platform. Final dose was calculated using the Eclipse AAA algorithm. Initial verification of delivery accuracy was measured using OSLDs on a solid water phantom of varying thickness. Results: A comparison of DVH curves demonstrated that dynamic couch motion irradiation was sufficiently approximated by static control points spaced in intervals of less than 2cm. Optimized MLC motion decreased the average lung dose to 68.5% of the prescription dose. The programmed irradiation integrating coordinated table motion was deliverable on a TrueBeam STx linac in 6.7 min. With the couch translating under an open 10cmx20cm field angled at 10°, OSLD measurements along the midline of a solid water phantom at depths of 3, 5, and 9cm were within 3% of the TPS AAA algorithm with an average deviation of 1.2%. Conclusion: A treatment planning and delivery system for Trajectory Modulated Arc Therapy of extended volumes has been established and experimentally demonstrated for TBI. Extension to other treatment techniques such as TLI and CSI is readily achievable through the developed platform. Grant Funding by Varian Medical Systems.

60 APPLIED LIFE SCIENCES↗

TU-F-CAMPUS-T-03: A Novel Iris Quality Assurance Phantom for the CyberKnife Radiosurgery System

Purpose: A novel CCD camera and conical scintillator based phantom that is capable of measuring the targeting and field size accuracy of a robotic radiosurgery system has been developed. This work investigates its application in measuring the field sizes and beam divergence of the CyberKnife variable aperture collimator (Iris). Methods: The phantom was placed on the treatment couch and the robot position was adjusted to obtain an anterior -posterior beam perpendicular to the cone’s central axis. The FWHM of the 12 Iris apertures (5, 7.5, 10, 12.5, 15, 20, 25, 30, 35, 40, 50, and 60 mm) were measured from the beam flux map on the conical scintillator surface as seen by the CCD camera. For each measurement 30 MU were delivered to the phantom at a dose rate of 1000 MU/min. The measurements were repeated at 4 SAD distances between 75 and 85 cm. These readings were used to project the aperture size as if the flux map on the scintillator were located 80 cm from the source (SSD). These projected FWHM beam diameters were then compared to the commissioning data. Results: A series of 12 beam divergence equations were obtained from the 4 sets of data using linear trend lines on Excel scatter plots. These equations were then used to project the FWHM measurements at 80 cm SSD. The average aperture accuracy for beams from 5 through 40 mm was 0.08 mm. The accuracy for the 50 and 60 mm beams were 0.33 and 0.58 mm when compared to film commissioning data. Conclusion: The experimental results for 10 apertures agree with the stated Iris accuracy of ±0.2 mm at 80 cm SAD. The results for the 50 and 60 mm aperture were repeatable and can serve as a reliable trend indicator of any deviations away from the commissioning values. Brett Nelson is President/CTO of Logos Systems.

60 APPLIED LIFE SCIENCES↗