Abstract: The present invention relates to a cell lysis or electroporation device comprising at least one pyroelectric material.

Abstract: The invention relates to a reagent and a process for the identification and counting of biological cells in a sample. This reagent comprises a cell lysing agent selected from at least one detergent in a concentration capable of specifically lysing a given type of cells in the sample, and a stain capable of marking the intracellular nucleic acids of the remaining unlysed cells. Application in particular for the identification and counting of cells using an automated analysis system based on flow cytometry.

Abstract: Method and apparatus for controlling acoustic treatment of a sample including a liquid. A processing volume in which the sample is acoustically treated may be controlled, e.g., by positioning a suitable element so as to reduce and/or eliminate a headspace at a sample/gas interface. An interaction between the acoustic energy and the sample may be controlled, e.g., by using an energy director positioned at least partially in the sample that helps to reduce splashing or other sample ejection that would otherwise occur.

Abstract: The instant invention provides an economical flow-through method for determining amount of target proteins in a sample. An antibody preparation (whether polyclonal or monoclonal, or any equivalent specific binding agent) is used to capture and thus enrich a specific monitor peptide (a specific peptide fragment of a protein to be quantitated in a proteolytic digest of a complex protein sample) and an internal standard peptide (the same chemical structure but including stable isotope labels). Upon elution into a suitable mass spectrometer, the natural (sample derived) and internal standard (isotope labeled) peptides are quantitated, and their measured abundance ratio used to calculate the abundance of the monitor peptide, and its parent protein, in the initial sample.

Abstract: A device for sample tissue disruption and/or cell lysis comprising: a piezoelectric material; and at least a second material in contact with the piezoelectric material; and wherein the second material has an uneven surface on an opposite side to that in contact with the piezoelectric material. The device may be made by assembling at least three layers and membranes for the valves and pumps. The piezoelectric material is actuated by an external voltage source to generate cavitation, which disrupts tissue and/or lyses cells, in particular by a modulated alternative external voltage. The invention further provides a method of disrupting tissue and/or lysing cells in a device. Also provided is a piezoelectric device comprising a piezoelectric material in contact with a second material, and wherein the second material has an uneven surface on an opposite side to that in contact with the piezoelectric material.

Abstract: Apparatus for microbial disruption, comprises a cylindrical transducer having a beat conducting, solid transmission layer disposed along at least a part of the inner longitudinal surface thereof, said transmission layer or said transducer and transmission layer defining a channel for receipt of a liquid sample, and means capable of delivering an alternating potential to said transducer at a frequency and voltage exciting resonance and inducing cavitation in said liquid sample.

Abstract: The present invention relates to the field of light induced therapy. The invention relates more particularly to a method of modulating receptor function of cells having receptor proteins, said method comprising illuminating the cells with light in the wavelength interval of 250-305 nm or with light having longer wavelengths that by means of non-linear processes and/or multiphoton excitation promotes the same electronic transitions as light in the wavelength interval of 250-305 nm to modulate said receptor function.

Abstract: The invention is a procedure for measuring the binding of an entity (ligand) to a surface by using a hapten-conjugated version of the ligand (hapten-ligand). An excess of the hapten-ligand is presented to the binding surface and excess (unbound) hapten-ligand is washed off. Bound hapten-ligand is then solubilized (removed) and applied to a membrane support or separated by electrophoresis and applied to a membrane support. Known amounts of hapten-ligand are similarly applied to the membrane, to provide for hapten-ligand standards. The membrane-bound hapten-ligand is detected by application of an enzyme-conjugated antibody to the hapten; or by application of an antibody to the hapten followed by application of an enzyme-conjugated antibody to the anti-hapten antibody. The resultant membrane-associated enzyme is detected and quantitated by the application of a color or light-producing substrate which reacts with the enzyme.

Abstract: A differential extraction system is directed to a microfluidic-based integrated cartridge to automate differential extraction of specific cell types within a mixed sample. The integrated cartridge includes a sonication module for selective cell lysis, separating means to eliminate centrifugation, high surface area pillar chip modules to purify DNA from a cell lysate, and microfluidic circuitry to integrate the steps in an automated platform.

Abstract: A biochip (100) for lysing and/or cell separation is formed to provide a sealed chamber for biological fluid. A conductive layer (140) bonded between upper (130) and lower (150) insulating layers is etched to form a microfluidic channel (250) between two electrodes (190, 200). The microfluidic channel connects a fluid inlet (11) and fluid outlet (120). The electrodes (190, 200) form an un-even electric field in the channel (250) to generate a dielectrophoretic force on the cells/particles within the sample fluid. A voltage source applies a suitable voltage to separate and/or lyse cells within the fluid.

Abstract: A sonication apparatus is directed to a microfluidic-based system to automate differential extraction of specific cell types within a mixed sample. The microfluidic-based system includes a sonication module for selective cell lysis, separating means to eliminate centrifugation, high surface area pillar chip modules to purify DNA from a cell lysate, and microfluidic circuitry to integrate the steps in an automated platform.

Abstract: The present invention relates to systems for releasing genetic materials from a solid medium. The present invention also relates to methods for releasing genetic materials from a solid medium. The present invention further relates to methods for isolating genetic material from a biological sample.

Abstract: A composition is disclosed with the composition comprising a mixture of natively glycosylated mammalian biological molecules produced by electromagnetically stimulating living mammalian cells.

Abstract: The invention relates to a method for the non-invasive selection of single living cells under gentle conditions from mixtures of cells or cell cultures with respect to a specific production performance. To this end, the concentration of substances produced by the individual cells which become enriched at the cell membrane, such as reporter gene products (GFP) or specifically secreted products, such as antibodies, is determined by fluorescence-microscopic detection methods.

Abstract: A device for measuring an extracellular potential of a test cell includes a substrate having a well formed in a first surface thereof and a first trap hole formed therein. The well has a bottom. The first trap hole includes a first opening formed in the bottom of the well and extending toward a second face of the substrate, a first hollow section communicating with the first opening via a first connecting portion, and a second opening extending reaching the second surface and communicating with the first hollow section via a second connecting portion. The first connecting portion has a diameter smaller than a maximum diameter of the first hollow section, greater than a diameter of the second connecting portion, and smaller than a diameter of the test cell. The device can retain the test cell securely and accept chemicals and the test cell to be put into the device easily.

Abstract: The present invention makes use of resonant acousto-EM energy (alone or in combination with resonant acoustic energy) applied to biologic structures for the disruption of at least one function of the biologic structure. In particular, the invention provides methods of generating acousto-EM energy in biologic structures such as virus, bacteria, fungi, worms and tumors for the disruption of these structures.

Abstract: Systems, devices, and methods for releasing one or more cell components from a photosynthetic organism. A bioreactor system is operable for growing photosynthetic organisms. Some of the methods include contacting the photosynthetic organism with an energy-activatable sensitizer, and activating the energy-activatable sensitizer, thereby releasing a cellular component from at least one of, for example, a membrane structure, tubule, vesicle, cisterna, organelle, cell compartment, plastid, or mitochondrion, associated with the photosynthetic organisms.

Abstract: A method for the physical and biological treatment of an organic effluent, which is decomposed by microorganisms in a biological treatment zone, includes recovering a fraction of the biomass derived from the biological treatment, filling a compression reservoir with the biomass fraction, isolating the compression reservoir, and subjecting the filled compression reservoir to a very high pressure so to destroy or damage the microorganisms in the biomass fraction. Thereafter, the compression reservoir is returned to atmospheric pressure and the treated biomass fraction is reinjected into the biological treatment zone or into another biological treatment zone.

Abstract: The present invention relates to a method by which a controlled source of heat (preferably a laser 3) generates a pulsating vapour bubble 4 in an enclosed liquid. The pulsating rate (the frequency) is preferably in the ultrasonic region, whereby cavitation occurs in the liquid. The cavitation effect will cause disruption of suspended components such as cells or bacterial spores. The ultrasonic disruption of cellular components is known as lysing by sonication.

Abstract: A device and methods for monitoring status of at least one cell, wherein the cell has a membrane forming a substantially enclosed structure and defining an intracellular space therein. In one embodiment of the present invention, the device includes a first substrate having a first surface and an opposite second surface, a second substrate supported by the first substrate, the second substrate having a first surface, an opposite second surface, a body portion between the first surface and the second surface, a first side surface and an opposite second side surface, wherein the body portion defines a first passage between the first side surface and the second side surface and an opening on the first surface of the second substrate and in fluid communication with the first passage, and sidewalls positioned above the first surface of the second substrate.

Abstract: Microfluidic systems and methods are disclosed which are adapted to transport and lyse cellular components of a test sample for analysis. The disclosed microfluidic systems and methods, which employ an electric field to rupture the cell membrane, cause unusually rapid lysis, thereby minimizing continued cellular activity and resulting in greater accuracy of analysis of cell processes.

Abstract: A cell lysis device for lysing cells or viruses, comprising a cell lysis tube having a sample inlet; a pump connected to the cell lysis tube for transferring a sample into the tube; a sealing unit for reversibly sealing a specific region of the tube; and a laser source for generating a laser is provide. Further, a method of lysing cells or viruses using the cell lysis device is provide. The method comprises introducing a sample containing cells or viruses and optionally magnetic beads to the cell lysis tube through the sample inlet; transferring the sample to a specific region in the cell lysis tube by means of the pump; temporarily sealing the region of the cell lysis tube where the sample is placed with the sealing unit; irradiating the sample with the laser; removing the sealing unit from the cell lysis tube; and discharging the sample from the cell lysis tube by means of the pump.

Abstract: A method of perforating a membrane 10 is presented. The method comprises: bringing a membrane-denaturing substance into contact with or close proximity to at least a site of said membrane 10, said substance inducing a membrane-denaturing reaction by a stimulus; providing said stimulus to said substance so as to denature said membrane 10; perforating said membrane with a membrane-destroying member 1. The stimulus is carried through said membrane-destroying member 1. The present invention eliminates the influence of membrane-denaturing agent to the substance to be injected into the cell. The present invention also enables applying the stimulus locally, with a simple construction.

Abstract: Treatment and/or prophylaxis, in mammalian patients, of neurodegenerative and other neurological medical disorders is effected by administering to the patient effective amounts of apoptotic bodies and/or apoptotic cells, preferably those derived from the patient's own white blood cells, e.g. by extracorporeal treatment of the patient's blood cells to induce apoptosis and administration of the apoptotic bodies and/or cells so formed to the patient.

Abstract: A method is presented for killing a first population of cells within a mixture of viable cells by providing the mixture of viable cells, contacting the cells with a label, illuminating a portion of the mixture, capturing an image of multiple cells in the illuminated portion of the mixture, determining at least two dimensional coordinates of one or more cells of the first population of cells using the first captured image, and applying a lethal dose of energy to said dimensional coordinates of the one or more cells at a first focal plane.

Abstract: The present invention provides a method and apparatus for selectively destroying dividing cells in living tissue formed of dividing cells and non-dividing cells. The dividing cells contain polarizable intracellular members and during late anaphase or telophase, the dividing cells are connected to one another by a cleavage furrow. According to the present method the living tissue is subjected to electric field conditions sufficient to cause movement of the polarizable intracellular members toward the cleavage furrow in response to a non-homogenous electric field being induced in the dividing cells. The non-homogenous electric field produces an increased density electric field in the region of the cleavage furrow. The movement of the polarizable intracellular members towards the cleavage furrow causes the break down thereof which results in destruction of the dividing cells, while the non-dividing cells of the living tissue remain intact.

Abstract: The present invention comprises devices and methods for performing channel-less separation of cell particles by dielectrophoresis, DC high voltage-pulsed electronic lysis of separated cells, separation of desired components from crude mixtures such as cell lysates, and/or enzymatic reaction of such lysates, all of which can be conducted on a single bioelectronic chip. A preferred embodiment of the present invention comprises a cartridge (10) including a microfabricated silicon chip (12) on a printed circuit board (14) and a flow cell (16) mounted to the chip (12) to form a flow chamber. The cartridge (10) also includes output pins (22) for electronically connecting the cartridge (10) to an electronic controller. The chip (12) % includes a plurality of circular microelectrodes (24) which are preferably coated with a protective permeation layer. Specific cells from various cell mixtures were separated, lysed, and enzymatically digested on the chip.

Abstract: An improved reagent system for preparation of cells for flow cytometry having a physiologically compatible salt solution composition including a lysing agent, an agent for minimizing white blood cell lysis, and optionally a preservative.

Abstract: The present invention relates to a method for disrupting cells by subjecting the cells to ultrasonic energy in the absence of beads. The present invention also relates to the enhancement of cell disruption methods using ultrasonic energy by reducing the surface tension of the liquid in which the cells are located.

Abstract: A method of determining information about cell viability and other characteristics relating to cell membrane permeability is disclosed. The method involves determining the effect of a cell on current flow and relating that effect to a known standard which standard may be a known healthy cell and thereby deducing the viability of the cell being tested. The cells being tested can be subjected to different environmental conditions such as surrounding chemicals, temperature, pH and pressure to determine the effects of such conditions on cell viability and/or cell permeability. The cell being tested can be in a cell suspension, grown on s ubstarte, in tissue in vitro or in tissue in vivo. The method provides substantially instantaneous results and need not include the use of dyes or other markers.

Abstract: A device for lysing components (e.g., cells, spores, or microorganisms) of a fluid sample comprises a cartridge having a lysing chamber for receiving the sample and having at least one solid phase in the lysing chamber for capturing the sample components to be lysed. An ultrasonic transducer is coupled to a wall of the lysing chamber to transfer ultrasonic energy to the captured sample components.

Abstract: This invention relates to devices and methods for performing active, multi-step molecular and biological sample preparation and diagnostic analyses employing immunochemical techniques. It relates generally to bioparticle separation, bioparticle enrichment, and electric field-mediated immunochemical detection on active electronic matrix devices utilizing AC and DC electric fields. More specifically, the invention relates to devices and methods for sample preparation/manipulation, immunoimmobilization, and immunoassays, all of which can be conducted on one or more active electronic chip devices within a single system. These manipulations are useful in a variety of applications, including, for example, detection of pathogenic bacteria and biological warfare agents, point-of-care diagnostics, food or medical product quality control assays, and other biological assays.

Abstract: Microfluidic systems and methods are disclosed which are adapted to transport and lyse cellular components of a test sample for analysis. The disclosed microfluidic systems and methods, which employ an electric field to rupture the cell membrane, cause unusually rapid lysis, thereby minimizing continued cellular activity and resulting in greater accuracy of analysis of cell processes.

Abstract: Intracellular material is released from bacterial, yeast, plant, animal, insect or human cells by the application of a low voltage such as 1 to 10 V to a suspension containing the cells. The conditions may be selected such that DNA released from the cells is electrochemically denatured so as to be available for use in an amplification procedure.

Abstract: The invention relates to a method for selectively releasing an agent loaded into a red blood cell, comprising electrosensitizing the red blood cell by application of an electric field and subsequently disrupting the cell selectively using ultrasound.

Abstract: The present invention provides low cost microfluidic devices having embedded metal conductors. The devices of the invention comprise a electronic component comprising a substrate having a first surface, a layer of electrically-conductive material deposited on a portion of the first substrate surface, a first sublayer of electrically-insulating material deposited on the first substrate surface and on the layer of electrically-conductive material, a second sublayer of electrically-insulating material deposited on the first sublayer of insulating material, and a third sublayer of electrically-insulating material deposited on the layer of dielectric material, and a fluid-handling component having a contoured surface affixed to the electronic component. The devices of the invention are advantageously used for performing electric field lysis and the polymerase chain reaction. The invention further advantageously provides simple, low cost methods for fabricating such microfluidic devices.

Abstract: A method for controlling organisms which comprises growing, decreasing, activating or inactivating cells, bacteria, viruses or fungi at an N-surface or a P-surface of a ceramic which is formed by treating the ceramic by polarization; and a material for controlling organisms, a method for selective adsorption of proteins, a material for selective adsorption of proteins, a cement material for filling bones and dental applications and a biomaterial, in which the ceramic treated by polarization is utilized. By utilizing difference in properties among surfaces of the ceramic treated by polarization, growth, decrease, activation or inactivation of organisms such as cells, bacteria, viruses or fungi can be controlled. Therefore, the above methods and materials are useful in the medical, dental and biochemical areas.

Abstract: A process and apparatus for cell purification and ablation is disclosed. The present invention comprises a laser system which directs radiant energy at computer or manually selected individual cells thereby disrupting DNA, RNA and protein structure in those cells. The present invention produces a purified tissue section containing relatively intact DNA, RNA or protein from only the untreated cells. This purified sample is suitable for amplification of material by PCR or other techniques for the analysis of molecular genetic features in the selected cells of interest.

Abstract: A method for counting leukocytes which comprises liberating elastase from granulocytes contained in a specimen, adding an anti-granulocyte elastase antibody to the thus liberated elastase, measuring the antibody bonded to the elastase to thereby determine the concentration of the elastase, and then calculating therefrom the number of leukocytes contained in the specimen with the use of the ratio of the leukocyte count to the elastase concentration. This method makes it possible to conveniently and less expensively count leukocytes at a high accuracy comparable to the one established by using a conventional automatic blood cell counter, as the figure shows.

Abstract: The invention provides apparatus and methods for subsecond lysis of selected cells in a cell chamber using a voltage pulse of 10 ms to 10 &mgr;s in duration followed by nearly simultaneous collection of the lysed cellular contents into a capillary electrophoresis tube or other suitable micro-collection device. Cell chambers and capillary electrophoresis tubes configured with electrodes for performing the electrical lysis are described. The influence of variables that govern the rate of cell lysis, such as inter-electrode distance, pulse duration, and pulse strength are also described. The methods are illustrated using fluorophores that are loaded into a cell then collected following electrical lysis, separated by electrophoresis, and then detected by laser-induced fluorescence detection in a capillary electrophoresis system.

Abstract: The invention describes a method for the isolation of components from samples, particularly large molecular weight DNA from biological samples. The method involves the application of controlled oscillatory mechanical energy to the sample for short periods of time of about 5 to 60 seconds to lyse the sample and release the component(s) from the sample, followed by standard isolation methods. In preferred embodiments, the method includes the use of a spherical particle for applying the mechanical energy.

Abstract: An apparatus including at least one sonotrode (2) designed to generate ultrasound of variable power in at least one biological sample (5) containing cells to be lysed, the sample (5) being contained in at least one receptacle (4 or 10) such that the sonotrode (2) is in direct contact with the receptacle(s) (4). Also disclosed is a method for using ultrasound to lyse a biological sample (5) contained in a receptacle (4), which includes placing the receptacle (4) in direct contact with the sonotrode (2), and activating the sonotrode (2) for long enough to lyse the cells in the sample (5) but preserve the DNA and/or RNA molecules released for subsequent operations, e.g. amplification. The invention is particularly applicable in the discipline of molecular biology.

Abstract: A process and apparatus for cell purification and ablation is disclosed. The present invention comprises a laser system which directs radiant energy at computer or manually selected individual cells thereby disrupting DNA, RNA and protein structure in those cells. The present invention produces a purified tissue section containing relatively intact DNA, RNA or protein from only the untreated cells. This purified sample is suitable for amplification of material by PCR or other techniques for the analysis of molecular genetic features in the selected cells of interest.

Abstract: Efficient cell lysis in small samples, i.e., samples less than one milliliter, is achieved by exposing the sample to microwave radiation in the frequency range of 18 to 26 GHz. The sample containing cells is supported in a wave-guide cavity, and a microwave source provides microwave radiation to the input port of the wave-guide cavity. A computer controls the frequency and source power level of the microwave radiation produced by the microwave source. The computer also monitors the input power level of the microwave radiation at the input port by means of an input power measuring instrument, the output power level at the output port by means of an output power measuring instrument, and the temperature of the sample by means of a thermocouple. In this way, the computer can control the operating parameters to achieve efficient cell lysis.

Abstract: The invention concerns a method for treating an aqueous flow colonized by cells with a pulsed electric field applied to a flow, characterized in that the applied field is substantially parallel to the direction of flow and to its application to the transfer of nucleic acids (RNA, DNA, oligonucleotides) into cells, to the transfer of proteins to cells, to the extraction of cytoplasmic macromolecules and molecules contained in the cells, to cell fusion and the production of hybrids and/or to insertion of membrane proteins. It also concerns an electropulsing chamber, a method for destroying cells and a membrane permeabilization method.

Abstract: A microwave device has a monolithic microwave integrated circuit (MMIC) disposed therein for applying microwave radiation to a microfluidic structure, such as a chamber, defined in the device. The microwave radiation from the MMIC is useful for heating samples introduced into the microfluidic structure and for effecting lysis of cells in the samples. Microfabrication techniques allow the fabrication of MMICs that perform heating and cell lysing of samples having volumes in the microliter to picoliter range.

Abstract: The present invention involves methods and devices which enable discrete objects having a conducting inner core, surrounded by a dielectric membrane to be selectively inactivated by electric fields via irreversible breakdown of their dielectric membrane. One important application of the invention is in the selection, purification, and/or purging of desired or undesired biological cells from cell suspensions. According to the invention, electric fields can be utilized to selectively inactivate and render non-viable particular subpopulations of cells in a suspension, while not adversely affecting other desired subpopulations. According to the inventive methods, the cells can be selected on the basis of intrinsic or induced differences in a characteristic electroporation threshold, which can depend, for example, on a difference in cell size and/or critical dielectric membrane breakdown voltage.

Abstract: Cysteine-depleted CTL epitopes can elicit a stronger or more specific CTL response than the native, cysteine-containing CTL epitope of a disease associated antigen.

Abstract: Microfluidic systems and methods are disclosed which are adapted to transport and lyse cellular components of a test sample for analysis. The disclosed microfluidic systems and methods, which employ an electric field to rupture the cell membrane, cause unusually rapid lysis, thereby minimizing continued cellular activity and resulting in greater accuracy of analysis of cell processes.

Abstract: This invention provides a method and apparatus for selectively identifying, and targeting with an energy beam, specific cells within a mixed cell population, for the purpose of inducing a response in the targeted cells. Using the present invention, every detectable cell in a population can be identified and affected, without substantially affecting non-targeted cells within the mixture.