Abstract: The introduction of genetic material or molecules of biological interest into cells is a procedure with an increasing interest both for experimental and application purposes, so that electroporation is a widely used technique, but the electroporation of single adhering cells is still impaired. The present application describes an apparatus for the electroporation of any kind of cell adhering to a substrate at any stage of development, where an electrical signal can be driven and applied to a single adhering cell in culture in order to obtain its electroporation. The method to electroporate a single adhering cell with the apparatus of the invention is also described.

Abstract: The present invention relates to an apparatus for conducting a variety of assays for the determination of analytes in liquid samples, and relates to the methods for such assays. In particular, the invention relates to a single-use cartridge designed to be adaptable to a variety of real-time assay protocols, preferably assays for the determination of analytes in biological samples using immunosensors or other ligand/ligand receptor-based biosensor embodiments. The cartridge provides novel features for processing a metered portion of a sample, for precise and flexible control of the movement of a sample or second fluid within the cartridge, for the amending of solutions with additional compounds during an assay, and for the construction of immunosensors capable of adaptation to diverse analyte measurements.

Abstract: Electroporation on a plurality of samples of membranous structures is performed in an electroporation well plate that includes a frame that can hold a plurality of well strips to form a two-dimensional array of wells, and a set of well strips, the set containing strips that differ in the number of wells while having the same outer dimensions and hence being interchangeable, thereby allowing the user to select strips appropriate for a given electroporation procedure, and allowing the manufacturer to replace defective strips without rejecting an entire well plate when a small number of wells is found to be defective.

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: Electroporation is performed on a population of cells, liposomes, vesicles, or other membrane-encased structures with uniform results regardless of size variations within the population, by drawing the membrane-encased structures into micron-sized openings that contain paired electrodes. An electric potential is then imposed between the paired electrodes to permeabilize only that portion of each cell that extends into the openings and resides within the electric field focused in the area between the electrodes.

Abstract: A device for performing cell assays has at least one assay site. The site comprises a pair of stimulating electrodes and a cell confinement cavity for confining at least one cell in an extracellular fluid medium. The cell confinement cavity and the stimulating electrodes are arranged so that, in use, each cell is exposable to an electrical field generated by a potential difference applied across the stimulating electrodes. The stimulating electrodes are spaced a distance apart such that the potential difference can induce field stimulation of each cell and simultaneously be below the level that would result in electrolysis of the extracellular fluid medium.

Abstract: A micro-organ structure comprising at least two micro-organ portions formed from a tissue, in which said at lest two micro-organs are linked one to the other by means of a junction formed from said tissue of which the micro-organs were formed therefrom.

Abstract: An implantable medical device is provided for use with an external detector to detect an analyte in vivo. In one embodiment, the device consisting essentially of a substrate; a plurality of discrete reservoirs located in the substrate, each reservoir having at least one opening; a reacting component contained in each reservoir; and at least one non-degradable barrier layer covering each reservoir opening, the barrier layer being permeable to an analyte to be detected, wherein the reacting component remains inside the reservoirs and can react with the analyte to be detected, and wherein the device is adapted for implantation into the body of a patient.

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: The present invention provides an apparatus, comprising a first mechanical structure having a first rigid surface, an area of the first rigid surface having a nanostructured surface. The apparatus also includes a second mechanical structure having a second rigid surface and opposing the first mechanical structure. The second rigid surface is cooperable with the nanostructured surface such that a microscopic particle is locatable between the nanostructured surface and the second rigid surface.

Abstract: Electroporation is performed in a controlled manner in either individual or multiple biological cells or 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. In addition, a particular method and apparatus are disclosed in which electroporation and/or mass transfer across a cell membrane are accomplished by securing a cell across an opening in a barrier between two chambers such that the cell closes the opening. The barrier is either electrically insulating, impermeable to the solute, or both, depending on whether pore formation, diffusive transport of the solute across the membrane, or both are sought.

Abstract: A system for qualifying cells of a cell sample labeled with a magnetic or magnetizable moiety is provided. The system includes a cell sample holder for holding a cell of the cells and a first cell analyzer which includes a magnetic field source for applying a magnetic field to the cell and a sensor for qualifying and/or quantifying an effect of the magnetic field on the cell.

Abstract: The present disclosure relates to the delivery of electric pulses any organic or inorganic conductive material and/or any biological material and/or to cells in vivo, ex vivo or in vitro, for example for the electroporation of the cells, for the electrically mediated transfer gene transfer of nucleic acids into tissue cell using a pulsed electric field and/or for the electromanipulation, in general, of the cell membrane or of the cell inside. The electric pulse applicator for the treatment of a conductive material such as biological material allowing an electric field to be applied to said conductive material in such a way as to modify it properties, includes at least one electrode including a conductive main body and an electrically insulating coating intended to be introduced into and/or at the vicinity of the conductive material to be treated, and a pulse generator sending pulses to the electrodes having a slope (dE/dt) greater than 1015 V/m/s.

Abstract: A chamber that includes electrical contacts and a protective lid with a manually operated release is designed to receive a multi-well electroporation plate and to allow high-throughput electroporation on the well contents with minimal risk of electrical shock to the user and minimal opportunity for sample loss and contamination.

Abstract: This invention generally relates to devices and methods for transfection of living cells using electroporation, in particular high throughput microfluidic electroporation, and to therapeutic uses of the transfected cells.

Abstract: Systems, methods and apparatus provide flexible and efficient high throughput electroporation systems. An electrical pulse may be transmitted to any number of channels of a multi-channel sample plate. Drivers can provide the selection of which channels to transmit the electrical pulse. To provide efficient transitions between electrical pulses, discharge circuits provide efficient means achieve a desired voltage.

Abstract: The present invention relates to an electrofusion microelectrode made of a tube having a first proximal end and a second distal end. The tube has an electrically conductive coating on its exterior surface that extends continually from the first proximal end of the tube toward the second distal end of the tube. Also disclosed is an electrofusion microelectrode unit having an electrofusion microelectrode and a holding tool capable of receiving the electrofusion microelectrode at the second distal end of the tube. The present invention also relates to a system having two or more electrofusion microelectrodes of the present invention and to methods of manipulating cells and/or cellular components using the electrofusion microelectrodes, units, and systems of the present invention.

Abstract: The electroporation chamber and its related devices combine the features of an electroporation chamber that acts as a manifold for regulation of sample flow with those of a flow electroporation device to form a regulated flow electroporation device. The invention further comprises a novel regulated flow electroporation chamber that enables conditions in which a sample is uniformly processed in individual fractions or volumes in a fully closed (sterile) system.

Abstract: The present invention relate to methods and devices for holding a cell and positioning recording electrodes inside it.

Abstract: A pulse generator circuit may include a diode configured to operate as an opening switch, a tank circuit in series with the diode having an admittance that is switchable from a first value to a second value that is different from the first value, and a switching system configured to cause the tank circuit to switch between the first value and the second value. The diode may saturate in less than 100 nanoseconds. A saturable core transformer may operate as a switch that controls the opening of the diode. The pulse generator may generate a plurality of pulses, each having a length of no more than 3 nanoseconds and an amplitude of at least 1 kilovolt. Electrodes may be connected to the pulse generator to deliver the plurality of pulses to biological cells.

Abstract: Device for the measurement of biological activities and/or physiological magnitudes, includes a measuring cell, equipped with a chamber suitable to receive micro-organisms to be analyzed, and one or more probes opening into the chamber. The probes are connected to measuring resources and resources for processing the electrical signals emitted by the probes. The measuring cell includes a vertical well, equipped in its lateral wall with holes uniformly distributed around the well in order to allow the probes to open into the well. The probes are sealingly attached to the well, and rest in the support resources, and suitable to receive a cup whose cross section is homothetic to the section of the well. The cup is equipped on its lateral wall with holes which are located opposite to the holes of the well when the cup is positioned in the well.

Abstract: A pulse generator circuit may include a diode configured to operate as an opening switch, a tank circuit in series with the diode having an admittance that is switchable from a first value to a second value that is different from the first value, and a switching system configured to cause the tank circuit to switch between the first value and the second value. The diode may saturate in less than 100 nanoseconds. A saturable core transformer may operate as a switch that controls the opening of the diode. The pulse generator may generate a plurality of pulses, each having a length of no more than 3 nanoseconds and an amplitude of at least 1 kilovolt. Electrodes may be connected to the pulse generator to deliver the plurality of pulses to biological cells.

Abstract: The present invention relates to a method, a chip, a device, and a system for detection of biological particles. The method of the invention typically comprises collecting the biological particles from a gaseous sample, contacting the biological particles with a first liquid reagent, extracting biological material from the collected biological particles, and analyzing the biological material for the presence of a target nucleic acid sequence.

Abstract: This invention generally relates to devices and methods for transfection of living cells using electroporation, in particular high throughput microtluidic electroporation, and to therapeutic uses of the transfected cells.

Abstract: An object of the present invention is to provide a cell culture vessel which is simple in structure and easy to handle, and is capable of preventing damage to the cells when separated, promoting transport of nutrients and excretion of effete matter, and elevating the culturing efficiency improving effect by the structural features. In order to attain the above object, there is provided a cell culture vessel including a culture section provided with a plurality of projections having an equivalent diameter smaller than the cells to be cultured and the culture section side walls enclosing the culture section, wherein the distance between an arbitrary position on the culture section/side wall boundary line and the nearest projection is smaller than the diameter of the cells to be cultured. The effect of the projections in the vessel given to the cultured cells is enhanced.

Abstract: The present invention provides a method whereby, in transferring a gene into a cell by contacting the cell with a gene to be transferred into the cell in a container and then electrospraying a spray liquid free from the gene on the cell and gene in the container, the gene can be rapidly and conveniently transferred into the cell with high transfer efficiency while minimizing the degradation of the gene, and an apparatus therefor. A nozzle for electrospraying comprising a tube portion for applying a high voltage, which is made of an electrically conductive substance and located on the side of the spray liquid suction port, and another tube portion for spraying, which is made of an insulating substance and located on the side of the spray liquid ejection port. By using this nozzle, electrospraying can be performed while preventing a discharge phenomenon that causes degradation of the gene.

Abstract: Provided is a reaction treatment apparatus in which, in a case of mixing a plurality of solutions in a microchip used in a biochemical reaction system, an electric field generation area for changing solute concentration distribution in the solutions is provided in a solution upstream fluid path. Diffusion between the solutions is accelerated by bringing an area with a high solute concentration into contact with another solution. This may shorten a time required for mixing the solutions.

Abstract: The present invention relates to a method of introducing nucleic acids into cells by electroporation, comprising the step (A) of loading nucleic acids to the surface of an electrode; the step (B) of adhering cells on the obtained nucleic acid-loaded electrode surface; and the step (C) of applying electric pulses to the adhering cells. According to this method, not only efficient introduction of a gene into cells but also gene introduction at desirable timing and at desirable sites can be performed without damaging the adhering cells.

Abstract: An apparatus and method for the dissociation of soft proteinaceous tissue using pulsed rapid variable direction energy field flow fractionization is disclosed. The pulsed rapid disruptive energy field is created by the use of a probe which surrounds the soft proteinaceous tissue to be removed. Once the adhesive mechanism between tissue constituents has been compromised, fluidic techniques are used to remove the dissociated tissue.

Abstract: Analytes using an active assay may be detected by introducing an analyte solution containing a plurality of analytes to a lacquered membrane. The lacquered membrane may be a membrane having at least one surface treated with a layer of polymers. The lacquered membrane may be semi-permeable to nonanalytes. The layer of polymers may include cross-linked polymers. A plurality of probe molecules may be arrayed and immobilized on the lacquered membrane. An external force may be applied to the analyte solution to move the analytes towards the lacquered membrane. Movement may cause some or all of the analytes to bind to the lacquered membrane. In cases where probe molecules are presented, some or all of the analytes may bind to probe molecules. The direction of the external force may be reversed to remove unbound or weakly bound analytes. Bound analytes may be detected using known detection types.

Abstract: The present invention relates to an improvement in an apparatus for detecting chemotaxis of cells. It aims at providing a structure for detecting chemotaxis of cells at an elevated accuracy with the use of a microquantity of cells. That is to say, an object of the present invention is to provide an apparatus for detecting chemotaxis of cells by which cell injection and position control can be easily carried out while ensuring the prevention of unexpected migration of the cells definitely positioned in a well or the injected sample so that a stable concentration gradient due to the diffusion of the specimen can be maintained and which ensures further automated operation and controlling.

Abstract: An electroporator for high-throughput electroporation is constructed with a well plate in which each well has internal electrodes that extend beyond the opening of the well to form contact areas, either as horizontal platforms extending laterally from the well rims or as extended heights of thin electrode plates. The electroporator also includes a lid that contains circuitry and electrical contacts that mate with the exposed contact areas in the well plate. The interchangeability of lids allows the wells to be shocked according to different protocols.

Abstract: The electroporation chamber and its related devices combine the features of an electroporation chamber that acts as a manifold for regulation of sample flow with those of a flow electroporation device to form a regulated flow electroporation device. The invention further comprises a novel regulated flow electroporation chamber that enables conditions in which a sample is uniformly processed in individual fractions or volumes in a fully closed (sterile) system.

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: Electroporation is performed in a controlled manner in either individual or multiple biological cells or 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. In addition, a particular method and apparatus are disclosed in which electroporation and/or mass transfer across a cell membrane are accomplished by securing a cell across an opening in a barrier between two chambers such that the cell closes the opening. The barrier is either electrically insulating, impermeable to the solute, or both, depending on whether pore formation, diffusive transport of the solute across the membrane, or both are sought.

Abstract: A pulse generator circuit may include a diode configured to operate as an opening switch, a tank circuit in series with the diode having an admittance that is switchable from a first value to a second value that is different from the first value, and a switching system configured to cause the tank circuit to switch between the first value and the second value. The diode may saturate in less than 100 nanoseconds. A saturable core transformer may operate as a switch that controls the opening of the diode. The pulse generator may generate a plurality of pulses, each having a length of no more than 3 nanoseconds and an amplitude of at least 1 kilovolt. Electrodes may be connected to the pulse generator to deliver the plurality of pulses to biological cells.

Abstract: An apparatus and methods for monitoring the status of a cell that consumes oxygen. In one embodiment of the present invention, the method includes the steps of confining the cell in a sensing volume, measuring dynamically intracellular or extracellular signaling of the cell, and determining the status of the cell from the measured intracellular or extracellular signaling of the cell.

Abstract: Disclosed is a device for processing biological material which at least comprises a chamber at least closable to the outside and having an inner space for receiving the biological material. The chamber comprises at least one electrode placed in contact with the inner space for generating an electric field. Also disclosed is a method for processing biological material. The biological material is introduced into the inner space above and the electrode can generate an electric field after said biological material is introduced by applying voltage to said electrode and a further electrode in contact with the inner space. The chamber comprises at least one inlet line having at least one opening arranged close to the electrode. The biological material is almost completely rinsed out of the inner space after the electric field is generated, via a solution guided via an inlet line of the chamber along at least one electrode.

Abstract: In method of injecting a substance into a living cell having a cell membrane, the substance, the cell and a liquid are placed into a tapering passage. Energy is applied to the cell, thereby inducing poration. To sort cells, a cellular suspension is placed in a tapering passage, including a narrow end that defines an opening that has a dimension corresponding to a cell size. An acoustic wave is applied, thereby forcing cells having a cell size smaller than the selected cell size through the opening, with a portion of the cells having a cell size not smaller than the selected cell size not forced through the opening. To extract material from a cell, an electric field and an acoustic wave are applied, thereby causing the cell membrane to allow the material to pass out of the cell.

Abstract: Disclosed is a method for treating biological material via electrical current, in which the biological material is added to a small volume of a buffer solution having relative high ionic strength. A strong electrical field is generated in the buffer solution by a high voltage pulse having a preset duration. The biological material is added to at most 50 ?l of a buffer solution with an ionic strength of at least 100 mmol/l. By at least one voltage pulse having a preset duration of at least 10 ?s, an electrical field with a field strength of at least 1 kV/cm is generated in the buffer solution. The voltage pulse is hereby interrupted at least once for a duration of at least 100 ?s and is then again continued.

Abstract: An object of the present invention is to provide a cell culture vessel which is simple in structure and easy to handle, and is capable of preventing damage to the cells when separated, promoting transport of nutrients and excretion of effete matter, and elevating the culturing efficiency improving effect by the structural features. In order to attain the above object, there is provided a cell culture vessel including a culture section provided with a plurality of projections having an equivalent diameter smaller than the cells to be cultured and the culture section side walls enclosing the culture section, wherein the distance between an arbitrary position on the culture section/side wall boundary line and the nearest projection is smaller than the diameter of the cells to be cultured. The effect of the projections in the vessel given to the cultured cells is enhanced.

Abstract: (Problems) To provide a cell fusion chip, which allows to perform a cell fusion on a single chip, excels in the operating efficiency, does not require experience so much in cell feed or recovery, and has no risk of the contamination to the cells during the operation. (Means for Solving Problems) The present invention provides a cell fusion chip comprising: an isolation chamber for receiving isolated cells to be subjected to an fusion operation; a fusion chamber for fusing the cells together supplied from the isolation chamber; a culture chamber for culturing the cells fused in the fusion chamber; a first channel for connecting the isolation chamber and the fusion chamber; and a second channel for connecting the fusion chamber and the culture chamber, wherein the isolation chamber, the fusion chamber, the culture chamber, the first channel and the second channel are formed on a single chip.

Abstract: The invention relates to a container 20, 30 for receiving an aqueous solution, which is formed at least partially by an outer limit 21 forming an inner chamber 22, 32 for receiving the solution, and which comprises at least one area which acts as an electrode 25, 26, 33, 34 when an electric voltage is applied and a subsequent discharge occurs, wherein at least one electrode 25, 26, 33, 34 is made of a conductive synthetic material at least based on a plastic material which is doped with at least one conductive substance. A container 20, 30 of the above-mentioned kind is created this way, which is simple and economical to produce and also, for example, enables an efficient transfection of living cells by means of electroporation or an effective electrofusion.

Abstract: Methods for differentially identifying cells in an instrument employ compositions containing a combination of selected antibodies and fluorescent dyes having different cellular distribution patterns and specificities, as well as antibodies and fluorescent dyes characterized by overlapping emission spectra which form non-compensatable spectral patterns. When utilizing the compositions described herein consisting of fluorescent dyes and fluorochrome labeled antibodies with overlapping spectra that cannot be separated or distinguished based upon optical or electronic compensation means, a new fluorescent footprint is established. This new fluorescent footprint is a result of the overlapping spectra and the combined cellular staining patterns of the dyes and fluorochrome labeled antibodies chosen for the composition. The new fluorescent footprint results in histogram patterns that are useful for the identification of additional cell populations or subtypes in hematological disease.

Abstract: Methods, systems, and apparatus provide an accurate time constant for electroporation. A model voltage function is created (analog, digital, or combination) to provide a model voltage having the desired time constant. A voltage is applied to the sample. A comparing circuit compares the output voltage, which may be attenuated, to the model voltage and provides an output control signal. This output signal is used to modify a resistance in parallel with the sample, thereby altering the output voltage to approximate the model voltage, which has the desired time constant. In one aspect, the control signal may be used to turn on and off a transistor that is in series with a resistor in order to modify the parallel resistance.

Abstract: The invention provides sensor, preferably biosensor devices and method of fabrication. The devices have significant advantages over the prior art methods having compatibility with future trends in clinical diagnostics and chemical detection. The underlying principle involves the integration of nanometer diameter, micron long metal or semiconductor rods onto a substrate to form a suspended nanomechanical cantilevers. The cantilever rods are rigidly attached to the substrate on one or both ends, and resonate at a characteristic frequency depending on the diameter, length, and stiffness of the rod. The metal or semiconductor rods are integrated onto the substrate using electrofluidic or fluidic assembly techniques. A receptor coating is placed on the metal or semiconductor rods prior to or following rod alignment using self-assembly chemistries.

Abstract: The present invention is directed to devices and methods that apply ultrasonic energy for the purpose of inducing transfection and cell transformation. A sonoporation system in accordance with embodiments of the present invention includes an ultrasonic electrical energy generator connected to an ultrasonic transducer producing stress waves. The ultrasonic transducer is connected to a fluid containment tank configured to accept at least a portion of the ultrasonic transducer whereby the ultrasonic stress waves may be delivered into the fluid medium. A cell holder is configured to hold one or more cells desirable for transfection. A hydrophone may be electrically connected to an acoustic stress wave intensity detection circuit. A motion control system having an arm configured to receive one or both of the cell holder and the hydrophone is configured to provide motion of one or both of the cell holder and the hydrophone within the fluid medium.

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: Electroporation is performed on a population of cells, liposomes, vesicles, or other membrane-encased structures with uniform results regardless of size variations within the population, by drawing the membrane-encased structures into micron-sized openings that contain paired electrodes. An electric potential is then imposed between the paired electrodes to permeabilize only that portion of each cell that extends into the openings and resides within the electric field focused in the area between the electrodes.

Abstract: Disclosed is a sensor comprising a substrate, a source contact region, a drain contact region, and the gate oxide of a transistor. A gate electrode is disposed between the gate oxide and a detection electrode made of a nonconducting material. The contact area Asens between the gate electrode and the detection electrode is larger than the contact area Agate between the gate electrode and the gate oxide, whereby the receptor can be immobilized on the surface of the detection electrode in a technically simple manner while the small contact area Agate between the gate electrode and the transistor provides for high sensitivity for detecting the analyte. According to the inventive method for detecting at least one analyte, at least one analyte is brought into contact with a receptor immobilized at the detection electrode so as to modify the electrical charge at the surface of the detection electrode. The analyte is detected by detecting the modified voltage in the transistor.