Life Sciences, Biotech, and Healthcare Technologies

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Monodisperse microdroplet generation

The steady-state phenomenon generates thousands of microdroplets per second which is a problem when the stream of droplets needs to be slowed down or stopped. LLNL technology provides a method for generating and trapping microdroplets at a desired location and subsequently stopping the stream of microdroplets without droplet coalescence. These microdroplets can then be chemically reacted, heated, cooled, optically interrogated, sorted and analyzed for as long as desired before channel flow is restarted. By trapping microdroplets at desired locations, greater optical or electromagnetic interrogation can occur. These results in a more thorough analysis of target analytes as well as increases reliability for techniques such as quantitative PCR.

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Pumping and valving for microfluidic

Researchers at LLNL have created a new technology for performing pumping and valving operations in microfabricated fluidic systems. Traditional microfabricated devices have some disadvantages that defeat the advantages of miniaturization. For example, they require high power and voltage, and they need specific fluids to work properly and to be broadly applicable. The technology described here uses low power, high pressure approach to slide the device's polymer plug within its microchannel, in turn controlling complex fluidic processes on-chip. This method is more efficient than current micro pumping and valving technology and addresses unmet needs for a totally integrated on-chip microfluidic system.

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Polymer-based microfluidic system

Researchers at LLNL have invented a new method for forming microfluidic system platforms that allow the incorporation of various microfluidic devices into a single unit. The method involves creating channels, reservoirs, and ports using a polymer-based platform that allows for the interconnection of building blocks. Pre-fabricated structures such as T’s and elbows are used to interconnect the building blocks. Microdevices can be embedded or bonded to the platform, creating a versatile and easily scalable system with applications in a variety of settings. One advantage of the present invention is that it can integrate several functions into one system, including pumping, mixing, diluting, separating, filtering, sensing, etc.

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Selective Immobilization of Proteins
Researchers at LLNL have developed a new method to utilize highly selective molecular recognition events to attach proteins to any solid support through the C-terminus. The approach is based on the use of protein trans-splicing, which is a naturally occurring process similar to protein splicing with the difference that the intein (e.g., DnaE intein from Synechocystis sp. PCC6803) self-processing domain is split into two fragments (N-intein and C-intein). These two intein fragments are inactive individually, however, under appropriate conditions they can bind to each other with high specificity to form a functional protein-splicing domain. In the method described here, the C-intein fragment is covalently immobilized onto a glass surface through a PEGylated-peptide linker, whereas the N-…

This LLNL-developed invention is multiplexed and utilizes the Luminex bead-based liquid array, which contains 100 different unique beads. Oligonucleotide probes with sequences complementary to the target sequences are covalently coupled to these unique beads. These capture beads are mixed with viral samples obtained from the patient via cheek swabbing or a throat wash and subjected to PCR in a conventional thermocycler. The amplified target sequence is then hybridized to complementary capture oligonucleotide probes via forward biotinylated primers. If this bead-probe-amplicon unit contains the target nucleic acid, it will be bound by the reporter molecule and fluorescence will be detected by a flow cytometer. This multiplexed assay would thus be able to detect and identify respiratory…

Researchers at LLNL have developed a novel method to express and purify significant quantities of AMPs. AMP is fused to the N-terminus of a self-assembling protein called encapsulin from Thermotoga maritima, which forms protein cages with 60 monomer units. N-terminal fusion of the peptide to encapsulin results in encapsulation of the peptide within the protein cage, which prevents cytotoxicity of the peptide to the host and protects the peptide from host proteolysis, ultimately enabling high cellular production levels. In order to isolate the peptide from the protein cage, specific protease cleavage sites (e.g., TEV protease sites) are engineered into the encapsulin cage. Upon treatment with the appropriate protease, the peptide is released and can be isolated for a designated purpose…

LLNL’s carbon nanotube trans-membrane channels invention is a new class of nanopores that combines the best features of all three existing types of pores while substantially mitigating a number of shortcomings exhibited by each of these types of pores.

The method involves sonication of nanotube in presence of lipids, including but not limited to DOPC or DPhPC. One advantage of this structure is the extended stability at room and/or elevated temperature.

LLNL has developed a novel process of production, isolation, characterization, and functional re-constitution of membrane-associated proteins in a single step. In addition, LLNL has developed a colorimetric assay that indicates production, correct folding, and incorporation of bR into soluble nanolipoprotein particles (NLPs).

LLNL has developed an approach, for formation of NLP/membrane protein complexes by simultaneous co-expression of both apolipoprotein and target membrane protein in a cell-free protein synthesis system. This approach involves cell-free transcription/translation technology adapted to co-express both apolipoproteins and a target membrane protein. It is carried out in a single reaction chamber with cell extract, buffer, phospholipds, detergents and the…

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Solid-state node-based rapid thermal cycler

Solid-state distributed node-based rapid thermal cycler for extremely fast nucleic acid amplification (LLNL Internal Case # IL-12275, US Patent 8,720,209)

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Laser heating for PCR

Laser heating of aqueous samples on a micro-optical-electro-mechanical system (LLNL Internal Case # IL-11719, US Patents 8,367,976;

LLNL researchers have designed a synthetic, concatemeric bacterial expression vector that expresses a protein sequence that can be digested into a single peptide. The synthetic protein is designed to be secreted outside E. coli cells, and therefore can be purified using a His-tag from the cell supernatant (thereby reducing the need to lyse the cells for purification). The protein sequence is optimized for protein solubility, and the concatemeric peptide sequence is optimized for ionization using electrospray ionization (ESI), which is the most common ion source for proteomics. Taken together, the protein can be spiked into a complex mixture at very low levels as an internal control, and then digested and detected as single peptide species.

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sensing, tracking and actuating droplets

LLNL scientists have created a technology that utilizes electrical means, instead of optical methods, to (1) provide label-free detection of droplet morphology; (2) manipulate droplet position through trapping and actuation; (3) track individual droplets in a heterogeneous droplet population; and (4) generate droplets with target characteristics automatically without optical intercession. The technology includes the use of electrodes to assay droplet contents and/or to actuate the droplet to release it from the trap. The technology is useful to detect the presence of cells, nucleic acids, proteins, or solute concentrations in an array of retrievable, trackable, trapped droplets in a closed fluidic system.

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Droplet array

This device allows for observation of single cells encapsulated in droplets and provide the ability to recover droplets containing a cell of interest. This system provides the unique capability to monitor droplet contents from a few minutes to hours and overcome the limitations of the fluorescence activated cell sorting (FACS) in the purification of cell populations. The ability of this technology, to retrieve individual droplets, allows for a wide variety of downstream analyses, such as PCR, sequencing, chromatography and mass spectrometry (e.g. LC-MS or GC-MS), or other analytical techniques.

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Rapid separation

This invention consists of a functionalized membrane (e.g. polyethylene glycol (PEG)) and osmosis or electric potential as a driving force. The PEG membrane provides high biological particles separation and prevents sample for clogging due to the strong hydration of functional polymers layer and their resistance to protein adsorption. The device is comprised of three chambers: 1) a feed and blood cells chamber, 2) a viral or bacterial concentration chamber, and 3) a cell waste chamber (see Figure). Each chamber is divided by membranes with different pore sizes and filled with electrolyte or draw solution to transport target solutes.

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Functionally graded materials

This invention is an improved chromatography device that utilizes the concept of a functionally graded material (FGM) for separation of components. The technology consists of a device that contains a FGM that is patterned to have a gradient in material properties (e.g. chemical affinity, surface chemistry, chirality, pore size, etc.) normal to the direction of flow of the mobile phase. The mixture is carried through the FGM by a mobile fluid phase, and the fluid that emerges from the FGM has the different components separated from each other.

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solid-state real-time optical monitoring

The art described here incorporates a planar integrated optical system that allows for multiple biochemical assays to be run at the same time or nearly the same time. Briefly, each assay can include one or more tags (e.g. dyes, other chemicals, reagents) whose optical characteristics change based on chemical characteristics of the biological sample being tested.

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Microfluid partitioning

This technology describes a method for partitioning fluid into “packets” between polymeric sheets. The fluid to be partitioned is introduced between two polymeric layers or within a polymeric channel and the layers are sealed together to form an array or sequence of individual milliliter to picoliter samples as shown in figure below. This approach allows a continuous flow of samples through the system and minimizes reaction and processing times. The invention described herein provides several advantages for DNA (or RNA) detection over the approach of suspending droplets in an immiscible fluid. The partitioned packets can be used in any assay such as PCR, isothermal amplification or other types of chemical or enzymatic analysis.

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AMS

LLNL researchers have developed a variation of AMS technology that improves sample preparation, analysis, and cost for AMS. The device involves depositing liquid samples on an indented moving wire and passing the moving wire through a combustion oven to convert the carbon content of samples to carbon dioxide gas in a helium stream. The gas is then directed via a capillary to a high efficiency gas-accepting ion source for AMS analysis (see Figure). The device is also applicable to handling continuous flows of liquid enabling on-line real-time liquid chromatography. This technology allows for assessment of human-relevant exposure levels directly in humans and reduces extrapolation from animal and in vitro testing.

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Microfluidic ultrasonic particle separator

LLNL has invented a new high-throughput assay for sample separation that uses the vibrations of a piezoelectric transducer to produce acoustic radiation forces within microfluidic channels. The system includes a separation channel for conveying a sample fluid containing the different size particles, an acoustic transducer and a recovery fluid stream. The polymeric films containing the fluid would be sealed upon application of heat and further partitioned into individual microliter or picoliter samples. This approach would allow for the purification, separation, and fractionation of different types of particles (viruses, proteins, nuclei acids, etc.) from complex biological samples suspended in a sample fluid.

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Ultraviolet radiation detector

LLNL researchers have developed an apparatus capable of measuring and recording ultraviolet radiation that uses the Schottky diode/ZnSe/metal type UV sensor. This device can detect both UV-A (320-400nm) and UV-B(280-320nm) radiation. The present invention can also measure and accumulate doses with good sensitivity, and it can also store and make available the readings to be downloaded for a period of one year. The capability of the instrument to record and storage the data is provided by the integration of the single-chip MSP430-type computer produced by Texas Instruments. The microchip contains a built-in analog-to-digital converter, which allows substantial energy efficiency and long service time of the instrument without replacement of the power supply.

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isotachophoresis system

The invention developed by LLNL researchers proposes to use staged isotachophoresis to improve sample separation. One of the problems with isotachophoresis is that there is a tradeoff between the diameter of the separation column and the ability to isolate a species into a detectable band. For example, wider diameter channels run faster, but narrower channels provide better ability to isolate one species into a detectable band. This technology uses staged isotachophoresis in which larger-diameter channels flow into one or more sequential channels with a reduced diameter (Figure). Although the width of the interface between species remains fixed, by reducing the channel width, the deleterious effect of diffusion across the interface is reduced.

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Liquid and gel electrodes

LLNL researchers have devolved a technique to separate or purify samples using electrophoretic separation. This invention corrects the problem associated with pH changes by using the electrode, which contacts the sample, itself a high-conductivity electrolyte made of liquid or gel materials. This will keep the metal surface electrochemistry physically remote from the sample, while applying the necessary electric field. The technique can be applied within microfluidic and chip-based platforms. In addition, it can also enable transverse-mode free-flow isotachophoresis. The device can be used to separate and purify samples such as biomolecules, viruses, bacteria, and cells.

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Bioluminescent probes

Researchers at LLNL have developed a more efficient and cost-effective method and system for synthesizing a critical D-aminoluciferin precursor and related compounds. D-aminoluciferin is as active as luciferin and provides a free -NH2 group for functionalization to attach peptide sequences corresponding to the cleavage site of a protease. This allows for the synthesis of bioluminescent probes for the detection of various proteases as diagnostic tools for diseases. However, D-aminoluciferin does not survive the harsh deprotection conditions used in the solid phase synthesis. LLNL inventors describe an alternative solution phase synthesis of D-aminoluciferin for generating protease probes.

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Chip-based droplet sorting

LLNL's technology employs improved sorting strategies related to chip-based droplet sorting. This technology uses electromagnetic fields and non-contact methods to sort and identify monodispersed water-in-oil emulsion droplets in a microfluidic chip-based device. The system selects individual droplets from a continuous stream based on optical or non-optical detection methods as well as the droplets interactions with applied alternating electromagnetic fields. The droplets are then sorted to different channels which are created by the system’s array of electrodes. The system also provides for the fabrication of the array of electrodes that allow selection and diversion of one or more droplets from a continuous-flowing stream.

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Passive chip-based droplet sorting

Researchers at LLNL have developed a method to passively sort individual microdroplet samples of uniform size based on stiffness and viscosity. Unlike electrical or optical methods for droplet sorting, this apparatus does not require a measurement step. Instead, particle separation occurs through changes in shearing forces determined by the stiffness of the particles in the microdroplet sample and controlled by the particle’s flow rate. Forks diverge from the devices’ main flow channels into two separate flow channels.

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Isotachophoresis

LLNL researchers have created a method that uses isotachophoresis for the exclusion and or purification of nucleic acids. Isotachopheresis (ITP) is an electrophoretic separation technique that leverages a heterogeneous buffer system of disparate electrophoretic mobilities. The researchers created a transverse ITP system that offers high-throughput sample preparation as the amount of sample that can be processed is not limited by an initial confined sample plug as in a traditional ITP system. With a judicious choice of electrolytes and some knowledge of electrophoretic mobilities of the samples to be separated, the transverse ITP system can be used to isolate DNAs from the mixture of viruses, cells, and other electrolytes in a filter-free approach.

Proteins and other functional molecules can often be synthesized in significant quantities, but their purification presents challenges. Also, many chemical/biological sensor technologies require that a small number of nanometer-sized molecules be filtered prior to being exposed to molecular recognition chemistries. Existing methods of filtering molecules by size for purification or sensing (e.g., porous microcomposites) suffer from several drawbacks, including (1) insufficiently small pore size (2) insufficiently uniform pore size, or (3) lack of mechanical stability or other deficiency in delivery mechanisms for forcing Iiquid through the pores. Thus, there is a need for low-cost filters with uniform pore size, scalable between 1-100 nm for protein screening.

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Chip-based sequencing

The present invention uses magnetic fields to hold particles in place for faster DNA amplification and sequencing. This invention provides a method for faster DNA sequencing by amplification of the genetic material within microreactors, denaturing and de-emulsifying and then sequencing the material while retaining it in the PCR/sequencing zone by a magnetic field. Briefly, nucleic acid sequencing occurs on a microchip. The nucleic acids are next isolated and hybridized to magnetic nanoparticles or to magnetic polystyrene-coated beads and microreactor droplets are formed.

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Chip-based device

This invention is designed to sort and identify complex samples using parallel nucleic acid characterization. By isolating single or double stranded nucleic acids derived from complex samples, researchers can sequence previously unknown genetic material to identify novel viruses and organisms. The chip-based microfluidic system achieves this through microdroplet PCR amplification, electrophoresis analysis and, genomic sequencing all performed on an integrated coplanar microfluidic system (see Figure). The present invention provides the opportunity for the scalable mass production for parallel and inexpensive microfluidic analysis chips.

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Multiplexed photonic membranes

This technology is a photonic detection system developed by researchers at LLNL for the detection of biological or chemical threats with the intention of combining the collection, concentration and detection process onto a single platform. The present invention consists of a porous membrane containing flow-through photonic silicon crystals (see figure). Photonic crystals constitute an emerging alternative technology, due to their powerful light-confinement abilities which would enable direct refractive index measurements, but also empower technologies based on absorption and Raman spectroscopy.

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Separating particles from a sample fluid

The described invention is a miniature fluidic device for separating particles suspended within a liquid sample that is introduced into the interior volume of the device. The device uses laminar flow and a combination of gravity and acoustic, electrophoretic, dielectrophoretic, and diffusion-based processes in concert to separate the different particle types and allow them to be collected separately or passed selectively to a subsequent device. The particles may be biological in nature, such as cells, viruses, or bacteria, possibly in combination with non-biological particles.

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Lipid nanotube or nanowire

Researchers at LLNL have developed a nanotube sensor (single-walled or multi-walled carbon nanotubes) enclosed within a highly selective lipid bilayer that can detect variations in ion transport using signal amplification generated from the disruption of protein pores across the lipid layer. Changes in the device’s transistor current are recorded by an external circuit with high efficiency as a result of the direct interface between the ion flow channels and the reporting nanostructure. The main sensing element of the device, as shown in the figure, is a carbon nanotube field-effect transistor (FET) with a single-wall carbon nanotube (CNT) connecting the source (S) and drain (D) electrodes.

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NERVe System Design
The team’s prototype is intended to be safe, simple and easy to build, while still achieving the minimally required functionality necessary to treat patients with COVID-19. The ventilator has two functional air flow circuits: an inhalation and an exhalation circuit (Figure 1). The pressure in each circuit—Peak Inspiratory Pressure (PIP) and Positive End-Expiratory Pressure (PEEP)—are controlled by two high-accuracy back pressure regulators. Thus, the device operates in pressure-controlled CMV (continuous mandatory ventilation) mode, which appears to be the most commonly used configuration for late-stage COVID-19 patients who require manual ventilation.
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Area vs Concentration Graph

LLNL scientists developed novel hydrogels, which are biodegradable soft materials synthesized by a water-soluble polymer. Incorporating silver imparts antimicrobial activity to the material at low concentration compared to currently used silver nanoparticles. Our hydrogels are composed of silver ions instead of silver nanoparticles, which eliminates the toxicity concerns of modern silver hydrogels. The hydrogels may be applied directly as a topical treatment for burns and wounds or may be added to a bandage or wound dressing.

LLNL scientists have invented a method that is able to identify SE specific sequences that can be utilized as diagnostics markers. The method, called suppression subtractive hybridization (SSH), is a PCR-based technique that identifies restriction fragments that are present in the target strain but not other strains. A set of restriction enzymes specific to the target species isolates their associated restriction fragments which are then amplified via PCR. These PCR amplicons are then validated for specificity by testing with primer pairs from several types of phages for the target pathogen and its close relatives. In this validation test, only the specific target pathogen would pair with its known phage primers, isolating it with high specificity even among its close relatives.

LLNL scientists have developed a method to ensure the accuracy of that tomographic image by applying adaptive optics (AO) to OCT in a single instrument (AO-OCT). AO stabilizes the image being captured by the OCT device by utilizing a Hartmann-Shack wavefront sensor and a deformable mirror, a type of mirror designed to compensate for detected waveform abnormalities (such as ones caused by a twitching eye) to produce a constant waveform image resulting in a sharp instead of a blurry image. The lateral resolution of the image is further improved by means of a second deformable mirror. Together, the sensors and mirrors stabilize and sharpen the image using the same light source as the OCT, where the sensors "instruct" the mirrors to deform to reflect the best image, thus enabling in vivo…

LLNL researchers have discovered unique DNA signatures that can be used to identify, with high specificity, three such organisms with bio-weapon potential, including Yersinia pestis and Francisella tularensis (both Category A agents), and Brucella species (Category B agent). The DNA sequence information of a desired region of an organism unique to that organism is recorded, a DNA primer is used to amplify the target fragment using the polymerase chain reaction (PCR), and then a hybridization probe is used to increase the specificity of the detection process by marking the target. Combined, this process uniquely identifies a DNA signature of an organism.

The LLNL invention has two assay chambers wherein each chamber is comprised of another two chamber modules. This allows the device to process up to two assays per chamber module, or four total assays per biological sample. These two duplex assays are each fed by parallel interrogation ports while the device still maintains a small physical profile. Each port has its own LED for excitation, allowing the second assay to have particularly improved excitation. The excitation of both assay chambers is achieved by a much simplified and straightforward optical system, which lowers the size, complexity, and cost of the device while increasing reliability and performance.

LLNL researchers have developed a method to quickly and accurately identify the family of a virus infecting a vertebrate via PCR. Universal primer sets consisting of short nucleic acid strands of 7 to 30 base pairs in length were created to amplify target sequences of viral DNA or RNA. These primers can amplify certain identifying sequences of all viral genomes sequenced to date as well as numerous virus subgroupings. The PCR products are separated on a gel by gradient electrophoresis to identify the virus. Altogether, these primers can identify all 28 known virus families that affect vertebrates. Each strain or species of virus produces its own unique electrophoretic banding signature, allowing for easy and quick virus identification. Primer libraries will be updated as new virus…

LLNL scientists have developed a high-confidence, real-time multiplexed reverse transcriptase PCR (RT-PCR) rule-out assay for foot and mouth disease virus (FMDV). It utilizes RT-PCR to amplify both DNA and RNA viruses in a single assay to detect FMDV as well as rule out other viruses that cause symptoms in livestock indistinguishable from those caused by FMDV, such as Bovine Herpes Virus-1 (BHV-1, 2), Bovine Papular Stomatitis Virus (BPSV), Bovine Viral Diarrhea Virus (BVD), Bluetongue Virus (BTV), Swine Vesicular Disease Virus (SVD) and Vesicular Exanthema of Swine Virus (VESV). This multiplexed assay contains individual Luminex-based assays for each of those seven diseases. Each assay involves probe sequences that are complementary to the target pathogen nucleic acid. The target…

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side_view_of_one_channel

LLNL researchers have invented a system for identifying all known and unknown pathogenic or non-pathogenic organisms in a sample. This invention takes a complex sample and generates droplets from it. The droplets consist of sub-nanoliter volume reactors which contain the organism sized particles. A lysis device lyses the organisms and releases the nucleic acids. An amplifier then magnifies the quantity of available nucleic acids. Then, a fractionator liberates the nucleic acids from the droplets. Finally, a parallel analyzer identifies all the known and unknown pathogen or non-pathogenic organisms in the complex sample. This device functions with DNA or RNA samples.

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image_of_field_deployable_instrument

LLNL researchers have developed a portable device which analyzes one or multiple types of body fluids or gases to test for one or more medical conditions. A bodily fluid (such as blood, perspiration, saliva, breath, or urine) is put into a condenser surface and is then separated into both a primarily gas fluid component and a second one that is primarily liquid. These two samples from the same fluid or gas source are subjected to analysis by, in various combinations, five different instruments: a condenser, functionalized nanostructures, an optional volatilizer, a differential mobility spectrometer, and an optional biomarker analyzer. Each instrument provides a unique analysis of a physical or chemical element of the tested bodily fluid.

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figure_of_kiosk

LLNL scientists have created a standalone pathogen identifier that can be placed in public settings, such as in stores or on street corners. Not unlike an ATM in physical size, this kiosk will accept biological samples from an individual for multiplexed analysis. The sample collection process will be sufficiently simple such that anyone could begin the diagnostic process after making the appropriate payment via cash, credit, or debit cards. After the customer signs the appropriate disclaimers concerning diagnosis and liability, a sterile swab or collection tool or vial, viral transport media, instructions on collecting a sample, gloves, and antiseptic wipes would be dispensed to them. The customer then would select what pathogens they want to be screened for before the assay begins.…

LLNL scientists have developed a technology which fulfills this need. The LLNL technology itself is comprised of two elements which are to be embedded in a user's personal electronic device (e.g. cell phone, tablet device, pager, etc.). The first is a proximity monitor which transmits location and temporal data such as the distance between the user and a contagious individual and the duration of proximity. The second is a personal exposure notification which comes after the user has been positively diagnosed with a contagious disease by a healthcare provider. Information of their contagiousness is downloaded to a server and the user's device would then transmit exposure warnings to other individuals who have encountered the user and are deemed by the proximity monitor to be at high…

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scheme

LLNL scientists have developed a method to synthesize long DNA sequences of varying length starting from short oligos. Synthetic oligos are generated using bioinformatics tools by overlapping multiple small segments, such as 4-mers or 6-mers, derived from both strands of the source DNA strand. DNA polymerases fill the gaps between these short n-mers to create the new, longer DNA strand. This process can be repeated multiple times using same or different length n-mers until the DNA strand of user specific length is synthesized within the microwell where this reaction takes place. An alternative version of this method, which separates the groups of different length n-mers spatially into distinct wells prior to the polymerase reaction occurring, is to separate them temporally. By…

LNLL scientists have invented a method for multiplexed detection of PCR amplified products which can be completed in a single step. Highly validated species-specific primer sets are used to simultaneously amplify multiple diagnostic regions unique to each individual pathogen. Resolution of the mix of amplified products is achieved by PCR product hybridization to corresponding probe sequences, attached to unique sets of fluorescent beads in liquid. The hybridized beads are processed through a flow cytometer, which detects presence and quantity of each PCR product. The assay is optimized to allow for maximum sensitivity in a multiplexed format. A background PCR product is formed via background multiplex PCR amplification reaction using a control DNA sequence. Comparing the fluorescence…

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figure_of_system

LLNL scientists have developed a rapid parallel genetic profiling technology that can be used to detect an array of pathogens from a small, complex sample. Detectable pathogens by the LLNL technology include viruses, bacteria, protozoa, and other microbes. The device works by first splitting a given sample into millions of emulsified, encapsulated microdroplets each of which are then split once more and run through a parallel analysis consisting of both a genomic and a proteomic assay. The droplets within the assays are first run through a PCR process which amplifies even the smallest quantities of DNA or RNA for analysis. Finally, the droplets, now with sufficient quantities of DNA or RNA for analysis after PCR, are dissolved and run on an agarose gel via electrophoresis. Analysis of…

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image_of_the_device

LLNL scientists have developed a battery-powered device which is low-cost and multi-chambered for the extraction and amplification of nucleic acids from environmental, clinical, and laboratory samples via loop-mediated isothermal amplification (LAMP). This platform identifies pathogenic bacteria and assists in determining the optimal treatment plan. A multi-chamber amplification cartridge in the device includes necessary reagents, a sample collection and transfer unit, and a heating unit which also contains a timer to track reaction start and stop times. The device also has a fluorescence detection component as well which is used with isothermal amplification techniques utilizing colorimetric detection to identify the target pathogen. The heating unit contained within the device…

IMPLANTABLE NEUROMODULATION SYSTEM FOR CLOSED-LOOP STIMULATION AND RECORDING SIMULTANEOUSLY AT MULTIPLE BRAIN SITES (IL13065; PCT Application WO2017100649)

This technology relates to a modular system for deep brain stimulation (DBS) and electrocorticography (ECoG). The system has an implantable neuromodulator for generating electrical stimulation signals adapted to be applied to a desired region of a brain via an attached electrode array. An aggregator module is used for collecting and aggregating electrical signals and transmitting the electrical signals to the neuromodulator. A control module that communicates with the aggregator module is used for controlling generation of the…

SENSOR ARRAY AND APPARATUS FOR SIMULTANEOUS OBSERVATION OF TISSUE ELECTROPHYSIOLOGY, CONTRACTILITY, AND GROWTH (IL13165, Pending US patent application)

Cardiac toxicity is one of the major causes of drug candidate failure in clinical studies and is responsible for the failure in regulatory approval of drugs as well as the retraction of numerous drugs from the market. Critical to the success of early stage drug discovery is the ability to obtain high-quality and high-throughput data in a cost-effective manner. Predicting cardiotoxicity requires the ability to create cardiac tissues that mimic in vivo physiology at multiple scales, as well as the ability to record two key cardiac functions: tissue electrophysiology and contractility.

Described is an iCHIP (in-vitro Chip-…

HIGH DENSITY POLYMER-BASED INTEGRATED ELECTRODE ARRAY (IL11207, US Patent 7,035,692)

This invention is a high-density polymer-based integrated electrode apparatus that comprises a central electrode body and multiple arms extending from the electrode body. The central electrode body with multiple arms is comprised of a silicone material with metal features in the silicone material, which comprises electronic circuits.

An advantage of this design is increased density of electrodes to meet increased resolution for devices, such as artificial vision and hearing implants.

DEPOSITING BULK OR MICRO-SCALE ELECTRONICS (IL12387, US Patent 9,485,873 and US patent Application US2017/0013713)

This invention provides thicker electrodes on microelectronic devices using thermo-compression bonding. A thin-film electrical conducting layer forms electrical conduits and bulk depositing provides an electrode layer on the thin-film electrical conducting layer. An insulating polymer layer encapsulates a thin-film of electrically conducting layer and the electrode layer. Some of the insulating layer is removed to expose the electrode layer.

This technology is advantageous for long-term…

Adhesive Actuated Insertion Shank

RIGID STIFFENER-REINFORCED FLEXIBLE NEURAL PROBES, AND METHODS OF FABRICATION USING WICKING CHANNEL-DISTRIBUTED ADHESIVES AND TISSUE INSERTION AND EXTRACTION (IL12469, US Patent Application US2014/0378993)

This invention is superior to silicon based neural probes, as it reduces localized damage induced by post-insertion movements. It is also more useful than purely polymer based probes, which are unable to penetrate neural tissue and therefore require an initial incision in the form of additional surgery, which may result in more glial damage.

Additionally, this stiffening shank apparatus may be easily and efficiently fabricated in large…

SERPENTINE AND CORDUROY CIRCUITS TO ENHANCE THE STRETCHABILITY OF A STRETCHABLE ELECTRONIC DEVICE (IL11169, US Patents 7,265,298 and 7,871,661)

STRETCHABLE POLYMER-BASED ELECTRONIC DEVICE (IL11206, US Patent 7,337,012)

This invention has a central longitudinal axis and is a stretchable polymer body in a longitudinal direction. Within the polymer is a circuit line extending in the longitudinal direction with an offset component that is at an angle to the longitudinal direction, forming a serpentine shaped circuit, allowing the…

MULTI-ELECTRODE NEURAL PROTHESIS SYSTEM (IL12575, US Patent Application US2016/0030753)

This invention entails a hermetically sealed electronics package of a multi-electrode neural prosthesis system, where the sealed enclosure communicates with external components via feedthroughs. The feedthrough density is increased, however, via signal demultiplexing prior to feedthrough submission.

This invention improves performance of long-term wireless, implantable devices by increasing signal density via demultiplexing. Additionally, this reduces the overall electronics package size, which is especially important in neural implants. The demultiplexing also reduces wireless data/power…

LLNL has developed specific technical approaches and methods to obtain proteomic information from various human tissue types (hair, skin, teeth, bone). These processes have been developed to maximize proteomic information recovery using liquid chromatography/mass spectrometry methods. LLNL has also developed software tools and processes to mine genetic databases and human genetic sequence data for specific mutations related to human identity that can be observed in the proteome. Finally, LLNL has established several advanced methods including biochemical processes that allow significant proteomic data to be obtained from short segments of single hairs as well as biochemical processes that enable both mitochondrial DNA (mtDNA) as well as proteomic information to be obtained from a…

LLNL Polyelectrolyte Enabled Liftoff (PEEL), is used to fabricate freestanding polymer films as thin as 10 nm that are capable of bearing loads ranging from milligrams to grams and deformations of up to forty percent (40%). PEEL employs robust, water-based, and self-optimizing surface chemistry to fabricate ultrathin films greater than 100 cm2 in area. The process is easily scalable in size and manufacturing quantity and applicable to a variety of polymeric materials.

The flexibility of PEEL is the key to its usefulness to industrial processes. It is scalable up to roll-to-roll level for high manufacturing volumes. It can use any kind of chemistry to generate membranes. Another significant benefit is that the film removal is water-based—it doesn’t need hazardous organic…

LLNL’s Forensic Science Center (FSC) is currently the only facility in the United States that is accredited to accept samples and analyze them for the possible presence of chemical weapons under the Chemical Weapons Convention. FSC scientists are global experts in chemical, nuclear, and biological counterterrorism and their work leads to innovative tools and therapies with biosecurity and public health applications. Working with LLNL computational biologists, and utilizing LLNL’s supercomputing infrastructure, FSC chemists recently identified a class of novel, neutral, cyclic oximes with increased blood brain barrier permeability serving as optimal antidotes against nerve agent poisoning. Cyclic oximes from this class show drastically increased permeability over HI-6 for the…

LLNL researchers have developed an acoustofluidic device design consisting of a silicon and glass chip bonded to a piezoelectric plate. The acoustic microfluidic chip design is optimized using numerical modelling for maximal pressure standing wave amplitude, and its unique configuration with subdivided channels enables high-throughput operation and customized placement of the acoustic pressure node. Experimental verification has demonstrated high-throughput size-separation of cells and viral particles, and label-free purification of biological samples.

Refer to the following publications for additional information on the research.

Efficient coupling of acoustic modes in microfluidic channel devices, Lab Chip, 2015, 15, 3192

Spatial tuning of…

LLNL has developed a brain-on-a-chip system with a removable cell-seeding funnel to simultaneously localize neurons from various brain regions in an anatomically relevant manner and over specific electrode regions of a MEA. LLNL’s novel, removable cell seeding funnel uses a combination of 3D printing and microfabrication that allows neurons from select brain regions to easily be seeded into an area approximately 2 mm across, with cell populations separated by less than 100 microns. LLNL’s MEA design is anatomically relevant, allowing cortical cells to be seeded around the periphery of the array and up to three other neuronal populations from interior regions of the brain in the central regions of the MEA.

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This technology uses microelectromechanical systems (MEMS), adaptive optics (AO), and optical coherence tomography (OCT) to produce 3-D retina images at the cellular level. AO compensates for optical aberrations by continuously sampling images, and rapidly compensating for these aberrations via a wavefront corrector. MEMS reduce the size and cost of the system without sacrificing speed or accuracy.

A select number of intracellular pathogens persist naturally in some amoebas and their cysts. LLNL has invented a technology that exploits this process to use amoeba cysts as natural containers for portable transport and long term storage. In addition, this novel encystment system incorporates sample purification and enrichment for clinical samples, taken in the hospital, lab, or any natural environment.

LLNL's multi-well plate cover penetration system is an array cutting and tape folding tool, based on 96-well, 384-well, and 1536-well geometries, that can be robotically operated and will cut, open, and fold inward the sealing tape so that samples can be subsequently aspirated without the need for human intervention to remove the seal which is an aerosol generating and contaminating process).

This technology uses either of two X-ray wave-front sensor techniques, Hartmann sensing and two-dimensional shear interferometry, both of which are capable of measuring the entire two-dimensional electric field, both the amplitude and the phase, with a single measurement. Capturing both the absorption and phase coefficients of the index of refraction can help to reconstruct the image. Measurement of the phase shift could enable the use of higher energy X-rays which would result in a lower amount of absorbed radiation to the patient, thereby reducing the potential damage to tissues. These wave-front sensors do not require a temporally coherent source and are therefore compatible with X-ray tubes and with laser-produced or X-pinch X-ray sources.

LLNL’s BioBriefcase is a compact and portable instrument capable of autonomously detecting the full spectrum of bioagents, including bacteria, viruses and toxins in the air. It uses the state of art technologies to collect, process, and analyze samples to detect, and identify genetic and protein signatures of bioagents.

The patented intracranial hematoma detection technology uses Micropower Impulse Radar (MIR). MIR uses short, high frequency electromagnetic pulses to obtain information in a non-invasive manner. Unlike ultrasound and other electromagnetic techniques, MIR can operate well through the skull, which is of great importance for intracerebral as well as epidural and subdural hematomas. The MIR hematoma detector has been studied in both laboratory model and human subject clinical experiments. The results have been reported in the Proceedings of SPIE 2005, Volume 6007.

Using various excitation wavelengths, a hyperspectral microscope takes advantage of autofluorescence and polarized light scattering from cellular components to obtain composite images that highlight their presence. The light collection efficiency is maximized to achieve image acquisition times and rates suitable for in vivo applications.

Lawrence Livermore National Laboratory scientists have developed a signal enhancing microchip apparatus and method that enhances a microfluidic detector's limits by magnetically focusing the target analytes in a zone of optical convergence. In summary, samples are associated with magnetic nanoparticles or magnetic polystyrene coated beads and moved down the flow channels until they are trapped in a magnetic trap-zone. The concentrated sample is then analyzed and provides up to a thousand fold signal enhancement for optical detection. Reaction volumes required are drastically reduced which improves sensor signal to noise ratio. (Sample amounts needed are one million times smaller than volumes used in current commercial instruments).

In addition to providing an enhanced signal…

LLNL researchers have combined a novel approach or using bioinformatics with cell-free expression to identify and characterize a class of proteins that kill Gram-positive bacteria with extremely high specificity. The class of proteins is collectively known as muramidases and possess bacterial lytic activity. Muramidases generally represent a potential class of novel antimicrobials for use against other Gram positive and, potentially, against Gram-negative infectious microorganisms. Strategies for effectively delivering these antimicrobial agents are being developed. These lytic proteins attack for the outside of the cells and circumvent mechanisms of antibiotic resistance that are found in many of the so-called drug-resistant superbugs. As such, they should be effective against these…

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Lawrence Livermore National Laboratory’s scientists have developed the Lawrence Livermore Microbial Detection Array (LLMDA), a technology enabling detection of bacteria, viruses and other organisms. This technology has shown value for applications in detection for product safety, diagnostics and bioterrorism events.

LLMDA contains probes fitted onto a one-inch by three-inch glass slide. Each probe tests for a particular sequence of DNA and small groups of probes can be used to check for specific bacteria or viruses up to the species level. The LLMDA can test for over 2,000 viruses and 900 bacteria. The newer version of the LLMDA will expand that capability to nearly 6,000 viruses and 15,000 bacteria as well as fungi and protozoa organisms. After DNA and/or RNA is extracted…

The VITA-D personal biodosimeter technology is designed to measure the vitamin D synthetic capacity of sunlight and/or artificial ultraviolet radiation in-situ, using the same photochemical process from which vitamin D3 is synthesized in human skin.

The innovators envision two embodiments of this technology:

  • Polymer films are doped with molecules of pro-vitamin D and are used as bio-analogue photo-registering elements. The dose measurement is based on changes in the transmission coefficient properties of the film. This can result in a disposable patch and reader technology.
  • Cholesteric liquid crystals (CLC) are doped with molecules of pro-vitamin D and the qualitative dose measurement is based on the color change, and the quantitative dose measurement…

The biotech industry aims to move towards an on-chip system for sample generation, amplification and detection of both DNA and RNA based organisms. LLNL has invented a new way of isolating samples in a system.

This invention enables creation of partitioned fluid "packets" between polymeric sheets for chemical separation, DNA amplification or PCR-based DNA detection. The polymeric films containing the fluid would be sealed upon application of heat and further partitioned into individual microliter or picoliter samples. This approach would allow a continuous flow of samples through the system and minimize reaction and processing times.

LLNL has developed a technology that provides near-instantaneous heating of aqueous samples in microfluidic devices. The method heats samples in a focused area within a microfluidic channel on miniaturized chips. The microwave heating device is composed of a waveguide or microstrip transmission line embedded in a microfluidic channel. Aqueous solution microwave heating allows extremely fast heat transfer for both heating and cooling.

LLNL is interested in developing a universal platform for the delivery and presentation of any protein antigen, including toxin, viral and bacterial proteins, with apparent concomitant adjuvant activity to enhance the host immune response.