Abstract: Described are methods for electrical treatment of biological cells, in particular using electrical field pulses, involving the steps: arrangement of the cells (1) on apertures (2) of a solid planar carrier element (3) which divides a measuring chamber (10) into two compartments (11, 12); and temporary formation of an electrical treatment field which permeates the cells, wherein an alternating-current impedance measurement takes place on the carrier element (3), and from the result of the alternating-current impedance measurement, a degree of coverage of the carrier element and/or healing of the cells after electrical treatment are/is acquired. Also described are devices for implementing the methods.

Abstract: Transfecting biology cells with nucleic acid molecules (DNA, siRNA) is an essential prerequisite in elucidating how genes function in complex cellular context and how their activities could be modulated for therapeutic intervention. Traditionally studies are carried out on a low throughput gene-by-gene scale, which has created a huge bottleneck in functional genomic study and drug discovery. Development of high-throughput cell transfection technology will permit functional analysis of massive number of genes and how their activities could be modulated by chemical or biological entities inside cells. This invention describes design, construction of device and apparatus for high throughput effective cell transfection. Procedures and protocols for using the device and apparatus are also described in the application. Novel methods of using the device in cell-based assays are also disclosed.

Abstract: A molecule introducing apparatus is provided with a power supply circuit 20 for supplying power, a step-up transformer 30 which is supplied with power from the power supply circuit 20 and which outputs a high voltage, a switching transistor 22 that blocks off and allows the supply of power from the power supply circuit 20 to the step-up transformer 30 and which generates an instantaneous high voltage, and a pair of electrode probes 50 for applying the instantaneous high voltage generated at the step-up transformer 30 to a predetermined region in a body. The molecule introducing apparatus oscillates DNA arranged outside a cell by the instantaneous high voltage so as to introduce the DNA into the cell.

Abstract: Disclosed is an apparatus and method for inserting one or several chemical or biological species into phospholipid containers that are controlled within a microfluidic network, wherein individual containers are tracked and manipulated by electric fields and wherein the contained species may be chemically processed.

Abstract: An electroporation device having a signal generating circuit (3) connectable at the output to electrodes (5) fittable to a substrate (35) having cells. The electrodes (5) produce an electric field which induces permeabilization of the cell membranes to facilitate introduction of substances into the cells. The amplitude of the signal (12) applied to the electrodes is closed-loop controlled on the basis of the impedance (Z(?)) of the substrate, so as to regulate the amplitude of the signal following a reduction in impedance caused by permeabilization of the cell membranes.

Abstract: An embodiment of the present invention provides a method for performing a lateral flow assay. The method includes depositing a sample on a test strip at an application region, detecting a first detection signal arising from the test strip in the first detection zone, and generating a baseline for the first measurement zone by interpolating between values of the detection signal outside of the first measurement zone and inside of the first detection zone. The method may include locating a beginning boundary and an ending boundary for the first measurement zone on the test strip. Additional detection zones having measurement zones may also be incorporated with the embodiment.

Abstract: The present invention relates to a substantially planar substrate for use in patch clamp analysis of the electrophysiological properties of a cell membrane comprising a glycocalyx, wherein the substrate comprises an aperture having a rim, the rim being adapted to form a gigaseal upon contact with the cell membrane, The invention further provides a method of making such a substrate and method for analysing the electrophysiological properties of a cell membrane comprising a glycocalyx.

Abstract: A biochemical analysis unit includes a substrate made of a material capable of attenuating radiation energy and/or light energy and formed with a plurality of holes, and a plurality of absorptive regions formed by forming an absorptive region in every hole. According to the thus constituted biochemical analysis unit, even in the case where the absorptive regions are formed at a high density, when a stimulable phosphor layer formed on a stimulable phosphor sheet is exposed to a radioactive labeling substance contained in the plurality of absorptive regions, electron beams (? rays) released from the radioactive labeling substance contained in the individual absorptive regions are reliably prevented from being scattered in the substrate and advancing to regions of the stimulable phosphor layer that should be exposed to electron beams released from absorptive regions formed in neighboring holes.

Abstract: An electrospraying apparatus and/or method is used to coat particles. For example, a flow including at least one liquid suspension may be provided through at least one opening at a spray dispenser end. The flow includes at least particles and a coating material. A spray of microdroplets suspending at least the particles is established forward of the spray dispenser end by creating a nonuniform electrical field between the spray dispenser end and an electrode electrically isolated therefrom. The particles are coated with at least a portion of the coating material as the microdroplet evaporates. For example, the suspension may include biological material particles.

Abstract: The present invention relates to a microfluidic system for processing a cell-containing liquid. The system includes a microfabricated substrate having an enrichment channel to prepare an enriched cell sample from the cell-containing liquid. A flow through member is in liquid communication with the enrichment zone. The flow through member substantially prevents cells of the cell-containing fluid from exiting the enrichment zone while allowing liquid of the cell-containing liquid to exit the enrichment zone. A gas actuator associated with the enrichment zone provides a gas pressure sufficient to move the enriched cell sample from the enrichment zone.

Abstract: A circuit for control of an output current in a multiple unit cell array includes an array of unit cells arranged in rows and columns. Each unit cell includes a column select transistor being adapted for control by a column selector and a row select transistor being adapted for control by a row selector. The column select transistor and the row select transistor are connected together in series to each other and between an output node and a first supply. A return electrode is provided to complete the circuit.

Abstract: Supports for immobilizing molecules, particularly biomolecules, methods of making such supports, kits, and biomolecular hybridization assay devices are disclosed. The methods, supports and devices include forming a triamine layer on at least a portion of a substrate and attaching a compound to a portion of the substrate.

Abstract: The present invention relates to a microfluidic system for processing a cell-containing liquid. The system includes a lysing zone to receive the cell-containing sample and a positioning element to position the cell-containing sample in a lysing position in the vicinity of a lysing mechanism. The lysing mechanism releases intracellular material, such as DNA or RNA, from the cells. In one embodiment, the lysing mechanism includes electrodes for generating an electric field sufficient to release intracellular contents from the cells. Alternatively, the lysing mechanism may lyse the cells using chemical, heat and/or ultrasonic techniques or any combination of these techniques.

Abstract: The present invention is directed to an electrofusion microelectrode used in the alignment, manipulation, fusion, or electroporation of cells. This device is particularly useful for transplantation of cells and cellular components.

Abstract: Drug candidate screening methods are applied to discover compounds with activity against ion channel targets. The method may include modulating the transmembrane potential of host cells in a plurality of sample wells with a repetitive application of electric fields so as to set the transmembrane potential to a level corresponding to a pre-selected voltage dependent state of a target ion channel.

Abstract: A micromachined cell lysis device with electrodes that are spaced by less than 10 ?m from one another. The cells are attracted to the space between the electrodes and then lysed.

Abstract: The present invention relates to methods and apparatus for the encapsulation of biologically-active substances in various cell populations. More particularly, the present invention relates to a method and apparatus for the encapsulation of biologically-active substances in various cell populations in blood by electroporation to achieve therapeutically desirable changes in the physical characteristics of the various cell populations in blood.

Abstract: Provided is an apparatus for detecting biopolymers (DNA) capable of total analysis including non-reacted samples without complicated operations such as washing. A DNA probe is fixed to one of electrodes and direct current voltage is placed between the electrodes, so that it becomes possible to separate complementary strand sample DNA and non-complementary strand sample DNA. By analyzing from a ratio in the whole reaction system, it is possible to obtain clearer results. Further, by using electrophoresis by gel together, it is possible to separate reacted samples and non-reacted samples to perform measurements therefor in the same reaction field.

Abstract: A sample separation apparatus including a porous, or rough, capillary column. The porous capillary column includes a matrix which defines pores, and may be formed rough surface of hemispherical grain silicon. The capillary column is defined in a surface of a substrate, such as silicon. The sample separation apparatus may include a stationary phase or a capture substrate disposed on the surfaces thereof. The sample separation apparatus may also include a detector positioned proximate the capillary column. A variation of the sample separation apparatus includes an electrode proximate each end of the capillary column. The sample separation apparatus may be employed to effect various types of chromatographic separation, electrophoretic separation, and analyte identification.

Abstract: The invention relates to a method for permeabilization of a cell structure consisting of a single cell, an intracellular structure or an organelle comprising the following steps: (a) microelectrodes, preferably two carbon fiber electrodes or hollow fiber electrodes, are provided, (b) the microelectrodes are connected to a power supply, (c) the electrodes, individually controlled by high-graduation micromanipulators, are placed close to the cell structure at an appropriate inter-electrode distance, and (d) a highly focused electric field of a strength sufficient to obtain electroporation is applied between the electrodes. The method may be used in order to transfer cell impermeant solutes, such as drugs or genes, into the cell structure or out of the cell structure, in biosensors, in the treatment of tumours and neurodegenerative diseases and in the study of biophysical processes.

Abstract: The present invention is directed to an electrofusion microelectrode used in the alignment, manipulation, fusion, or electroporation of cells. This device is particularly useful for transplantation of cells and cellular components.

Abstract: A ready-to-use electroporation cuvette is provided that includes a cuvette, first and second electrodes positioned within the cuvette and electroporation competent cells frozen in a suspension solution within the cuvette, wherein the electroporation cuvette is configured to permit electroporation of the cells when the cells are thawed. The electroporation cuvette may be sealed with a cap that may be color coded to aid the user.

Abstract: Assays for detecting molecular modifications such as phosphate modifications and the presence and/or activity of enzymes and other agents involved in facilitating or otherwise regulating such modifications.

Abstract: Electroporation is performed in a controlled manner in individual and multiple biological cells present in biological tissue by monitoring the electrical impedance, defined herein as the ratio of current to voltage in the electroporation cell. The impedance detects the onset of electroporation in the biological cell(s), and this information is used to control the intensity and duration of the voltage to assure that electroporation has occurred without destroying the cell(s). This is applicable to electroporation in general.

Abstract: A multiple electrode includes a plurality of micro-electrodes provided on a substrate, and a wiring portion for providing an electrical signal to the micro-electrodes or extracting an electrical signal from the micro-electrodes. Each micro-electrode has porous conductive material on its surface, and the impedance of the micro-electrode is 50 k? or less. Preferably, the porous conductive material is gold, and formed by the passage of current at a current density of 1.0 to 5.0 A/dm2 for 10 to 360 sec. The multiple electrode may include micro-electrodes provided on a substrate in the form of a matrix, a lead line connected to the micro-electrodes, and an electrical junction connected to an end of the lead line.

Abstract: The present invention relates to methods and apparatus for the encapsulation of biologically-active substances in various cell populations. More particularly, the present invention relates to a method and apparatus for the encapsulation of biologically-active substances in various cell populations in blood by electroporation to achieve therapeutically desirable changes in the physical characteristics of the various cell populations in blood.

Abstract: This new automatic bio-manipulation factory system for manipulating a single cell comprises: a bio-MEMS that transports, separates, and extracts a single cell for manipulation from a plurality of cells, controls the rotation-orientation of the extracted cell automatically, and enables the extracted cell to maintain a certain temperature; and a micro manipulation system that obtains vision information and manipulation information about the extracted single cell and the manipulation tool, provides such information to operator, manipulates the extracted single cell by transmitting the rotation-orientation feedback information and the cell manipulation order of the operator to the bio-MEMS, and manipulates the manipulation tool by transmitting the tool manipulation order of the operator to the manipulation tool. The conventional manual and individualized single cell manipulation can be automated and processed collectively by systematically combining the bio-MEMS with the micro manipulation system.

Abstract: A chamber for treating cells contained in a suspension in an electric field, the chamber having a vessel for holding the suspension and at least two electrodes with electrode surfaces that face each other. The suspension may be placed between the electrode surfaces and the electrodes are connectable to different poles of a source of voltage to generate an electric field between the electrode surfaces. The electrodes are laminate and continuous and are configured to generate a non-uniform field. Moreover, the electrodes have electrode surfaces made of electrically conductive material, wherein the electrode surface of at least one of the two electrodes is shaped such that a non-uniform field with several maxima distributed over the electrode surface may be generated between the two electrodes.

Abstract: The invention comprises an apparatus and a method of effecting localized electroporation in a relatively small target area and for introducing foreign matter into cellular material in which the target area is located. The cellular material may be in vitro, in ovo or in vivo.

Abstract: An electroporation chamber is provided wherein cells are borne by a fluid medium through a conductive mesh receiving electrode (with the mesh size being such that it allows passage of cells of the desired size/type) and into the chamber. The cells are then captured on a conductive mesh capturing electrode having a mesh size which retains the desired cells. The electrodes are then charged to effect electroporation or other operations within the captured cells between the electrodes. If desired, different fluid media may flow through the electrodes and chamber (and over the cells) during such activities. When operations are completed, fluid flow may be reversed to carry the treated cells back out of the chamber through the receiving electrode.

Abstract: The controllable electroporation system and method described herein allows control over the size, the number, the location, and the distribution of aqueous pores, thus increasing flexibility of use. The herein described system and method for controllable electroporation generally employs at least two actuating sub-systems and sub-processes. One sub-system and sub-process employs a relatively broad effect in order to weaken the membrane, a broad effect sub-system. Another sub-system and sub-process employs a relatively narrow effect in order to localize the position of the pore in the membrane, a narrow effect sub-system.

Abstract: Disclosed are devices for amplifying a preselected polynucleotide in a sample by conducting a polynucleotide polymerization reaction. The devices comprise a substrate microfabricated to define a sample inlet port and a mesoscale flow system, which extends from the inlet port. The mesoscale flow system includes a polynucleotide polymerization reaction chamber in fluid communication with the inlet port which is provided with reagents required for polymerization and amplification of a preselected polynucleotide. In one embodiment the devices may be utilized to implement a polymerase chain reaction (PCR) in the reaction chamber (PCR chamber).

Abstract: The manipulator includes a support and at least one member affixed to and extending away from the support. The member has at least two differentially activatable areas of conductivity. The members are configured to establish a first electromagnetic field in vivo between selected areas of conductivity sufficient to manipulate a molecule relative to a target tissue and to establish a second, typically higher, electromagnetic field sufficient to cause transient permeability of a cell membrane within the target tissue. Restraining means are also described for restricting movement of the members with relation to each other. One method of using the device is for enhancing the delivery of a molecule into a tissue site; another is for poration of the tissue alone or in combination with the migration. The target tissue may include a tumor, organ, or wound site.

Abstract: The present invention provides a method and apparatus adapted to facilitate the entry of a preselected molecule into the intracellular space of a cellular sample through the use of ions generated by a corona charge source. With the present method and apparatus, molecules are manipulated within cells and in the extracellular space surrounding the cells. Manipulation enhances the permeability of cell barriers to allow the subsequent introduction of molecules of interest into the interior of a cell.

Abstract: A system and method are described for electroporating a sample that utilizes one or more sets of electrodes that are spaced apart in order to hold a surface tension constrained sample between the electrodes. The first electrode is connected to the lower body of the system while the second electrode is connected to the upper body. Both electrodes are connected to a pulse generator. Each electrode has a sample contact surface such that the first electrode and the second electrode may be positioned to hold a surface tension constrained sample between the two sample contact surfaces and the sample may receive a selected electric pulse.

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: The invention relates to an affinity sensor for detecting specific molecular binding events, for use in the field of molecular biology, e.g., in medical diagnostics, especially in biosensor technology or in DNA microarreay tests. The aim of the invention is to provide an affinity sensor of this type for rapidity, sensitively, specifically, economically and routinely detecting the presence of molecules, especially bioactive molecules, and to provide special applications for an affinity sensor of this type. To this end, the affinity sensor consists of a support substrate which is provided with at least two electrodes. The electrodes are situated equidistantly from each other and cover an area on both sides, at least this area being provided for receiving immobilized specific binding partners which are capable of coupling complementary corresponding binding partners directly or with other specific binding molecules.

Abstract: The present invention is a composition for percutaneous administration suitable for electroporation. Percutaneous absorption of a drug and so forth in a composition for electroporation can be promoted by adding alkaline earth metal ions to the composition.

Abstract: An apparatus is provided for receiving and aligning a multiple electrode pair array and a multiple well plate used in electroporation. The apparatus includes a housing which includes a bottom housing portion and a pair of side housing portions which project upward from the bottom housing portion. A pair of board-reception structures are adjacent to the inner portions of the side housing portions. Multiple well plate engaging and disengaging means, supported by the housing, are provided for engaging a multiple well plate with a multiple electrode pair array and for disengaging a multiple well plate from a multiple electrode pair array. Preferably, the multiple well plate engaging and disengaging means include a multiple well plate lifting and lowering assembly. The multiple well plate lifting and lowering assembly can include lifting/lowering handles located outside the housing.

Abstract: Microchip devices and methods of use are provided for the controlled exposure of a secondary device. In one embodiment, the microchip device include a substrate, a plurality of reservoirs in the substrate, a secondary device in one or more of the reservoirs, at least one barrier layer covering each of the reservoirs to isolate the secondary device from an environmental component outside the reservoirs, wherein the barrier layer is impermeable to the environmental component, and a microprocessor-controlled actuation means for rendering the barrier layer porous or permeable to the environmental component to expose the secondary device to said environmental component. The secondary device can comprise a sensor or sensing component (e.g., a biosensor). In other embodiments, microchip devices are provided for the controlled exposure of a reacting component, such as an enzyme. In still other embodiments, sensors are provided on the device outside of the reservoirs.

Abstract: A single cell electroporation assembly and method involve delivery of a substance into a single cell. The substance is combined into an electroporation fluid and placed into a container having a distal opening. The distal opening of the container is placed in proximity to a target cell. Electrical pulses are delivered between a first electrode which is at least partially disposed in the container, and a second electrode outside the cell, the cell being positioned between the distal opening and the second electrode. The electrical pulses induce temporary formation of pores in the cell membrane and the substance enters the cell through the pores by passive diffusion or by active electrophoretic motion.

Abstract: The controllable electroporation system and method described herein allows control over the size, the number, the location, and the distribution of aqueous pores, thus increasing flexibility of use. The herein described system and method for controllable electroporation generally employs at least two actuating sub-systems and sub-processes. One sub-system and sub-process employs a relatively broad effect in order to weaken the membrane, a broad effect sub-system. Another sub-system and sub-process employs a relatively narrow effect in order to localize the position of the pore in the membrane, a narrow effect sub-system.

Abstract: A culture apparatus and cultivation method are provided which enable liquid materials such as cells or blood plasma to be evenly cultivated by making current density uniform.

Abstract: Apparatus for conducting electrophysiological measurements on cells wherein a measuring head is provided with at least one electrode in a common support for impaling the cells.

Abstract: A method for sustained intravascular delivery via electroporation is provided. The method is useful for delivery of therapeutic compositions such as antithrombotic and anticoagulant agents. The invention also provides a catheter apparatus for introducing a composition into at least one cell in a vessel in a subject.

Abstract: The present invention relates to methods and apparatus for the encapsulation of biologically-active substances in various cell populations. More particularly, the present invention relates to a method and apparatus for the encapsulation of biologically-active substances in various cell populations in blood by electroporation to achieve therapeutically desirable changes in the physical characteristics of the various cell populations in blood.

Abstract: An electrospraying apparatus and/or method is used to coat particles. For example, a flow including at least one liquid suspension may be provided through at least one opening at a spray dispenser end. The flow includes at least particles and a coating material. A spray of microdroplets suspending at least the particles is established forward of the spray dispenser end by creating a nonuniform electrical field between the spray dispenser end and an electrode electrically isolated therefrom. The particles are coated with at least a portion of the coating material as the microdroplet evaporates. For example, the suspension may include biological material particles.

Abstract: A device for manipulating a molecule in vivo relative to a target tissue includes at least one elongated member having at least two discrete and separately activatable electrodes separated by an insulating material interposed therebetween. The electrodes are configured to establish at least one of a first electromagnetic field between selected electrodes sufficient to manipulate a molecule relative to a target tissue and a second, typically higher-level, electromagnetic field sufficient to cause transient permeability of a cell membrane within the target tissue. A third electromagnetic field may also be applied to cause further translation of the molecule into an electropermeabilized cell and/or manipulated with respect to the tissue. Thus three-dimensional manipulation of the molecule relative to the target tissue may be effected to optimize a desired positioning thereof, such as entry into a cell.

Abstract: A method for cell and electrical therapy of living tissue including administration of exogenous cells into a region of injured tissue and application of electrical energy. In one application the combined cell and electrical therapy is applied in vivo to damaged heart tissue. In such applications, minimally invasive procedures are used to apply the cell therapy and the electrical therapy is provided via an implantable pulse generator. In one application an implantable pacemaker is used in the VDD mode with an atrioventricular delay kept relatively short when compared to the intrinsic atrioventricular delay.

Abstract: A method for cell and electrical therapy of living tissue including administration of exogenous cells into a region of injured tissue and application of electrical energy. In one application the exogenous cells are conditioned in vitro before their administration into tissue. In one application the combined cell and electrical therapy is applied in vivo to damaged heart tissue. In such applications, minimally invasive procedures are used to apply the cell therapy and the electrical therapy is provided via an implantable pulse generator. In one application an implantable pacemaker is used in the VDD or DDD mode with an atrioventricular delay kept relatively short when compared to the intrinsic atrioventricular delay.