Abstract: A method of treating biocells includes the steps of: a. providing biocells; b. applying at least one stressor to the biocells sufficient to cause nonlethal and reparable cell wall damage to the biocells, thereby putting the biocells in a catabolic state during which catabolic metabolic functions predominate over anabolic metabolic functions; and c. obtaining at least one product produced by the biocells during the catabolic state. In another embodiment, the method includes the steps of: a. providing biocells that are mammalian cells; b. applying at least one stressor to the biocells sufficient to cause nonlethal and reparable cell wall damage to the biocells, the reparable cell wall damage comprising openings that allow increased passage of materials through the cells walls; and c. inserting foreign DNA through the openings into the biocells.

Abstract: A method for introducing biologically active molecules into animal or human cells using an electric current includes suspending the cells and dissolving the biologically active molecules in a buffer solution including HEPES and at least 10 mmol×1?1 magnesium ions (Mg2+), the buffer solution having a buffer capacity of at least 20 mmol×1?1 ×pH?1 at a change in the pH from pH 7 to pH 8 and at a temperature of 25° C., and an ionic strength of at least 200 mmol×1?1. An electric voltage is applied to the suspension.

Abstract: The invention relates to a novel circuit arrangement for electrotransfection or electrofusion, which enables the transportation of DNA and/or other biologically active molecules to the nucleus of higher eukaryotic cells or the fusion of cells, independent of cell division and with reduced cell mortality.

Abstract: Antisense sequences, including duplex RNAi compositions, which possess improved properties over those taught in the prior art are disclosed. The invention provides optimized antisense oligomer compositions and method for making and using the both in in vitro systems and therapeutically. The invention also provides methods of making and using the improved antisense oligomer compositions.

Abstract: A plasma containing at least one reactive species are used to deliver agents into living cells or an extracellular matrix of the living cells. The method and system allows for treatment of the living cells without causing irreversible damage to the cell membranes of the living cells. The plasma is produced by dielectric barrier discharge.

Abstract: Embodiments of the invention are directed to a technique for electroporation that allows for a delivery of long electrical pulses of high magnitude in highly conductive buffers and minimizes damage to cells undergoing electroporation.

Abstract: The present invention relates to a device for treating biological cells in an object, the device comprising: —a single winding coil element; —an electrical generator connected to the single winding coil element, the single winding being configured to be positioned essentially around the object; wherein the electrical generator is configured to discharge into the single winding coil element so that the single winding coil element generates a short duration pulsed electromagnetic field by magnetic induction in the single winding coil element, the electromagnetic field having a field strength that is sufficiently high to affect, preferably increase the permeability of cell membranes and/or intracellular membranes of the biological cells contained in the object when in operation the object is placed inside the single winding coil element.

Abstract: Methods, tip assemblies and kits are provided for introducing material into cells. The tip assemblies include an attachment portion, a channel portion, and a constriction that function to reduce fluid pressure as a fluid passes through the constriction portion from the channel portion, whereby the tip assemblies form pores in the membranes of cells and introduce material into the cells. The material includes for example one selected from the group of: an inorganic compound, a drug, a genetic material, a protein, a carbohydrate, a synthetic polymer, and a pharmaceutical composition.

Abstract: A system and method are disclosed for extracting lipids from algal cells. In the method, lipids are extracted from algal cells by exposing the algal cells in an aqueous medium to an electric field sufficient to cause release of lipids from said cells. In the system, an electric field is formed between two electrodes connected with an electrical power supply and configured such that during use an aqueous medium containing the algal cells passes between the electrodes to extract lipids therefrom.

Abstract: A system for treating benign prostate hyperplasia (BPH) of a prostate. At least first and second mono-polar electrodes are configured to be introduced at or near a BPH tissue site of the prostate gland of the patient. A voltage pulse generator is coupled to the first and second mono-polar electrodes. The voltage pulse generator is configured to apply sufficient electrical pulses between the first and second mono-polar electrodes to induce electroporation of cells in the BPH tissue site, to create necrosis of cells of the BPH tissue site, but insufficient to create a thermal damaging effect to a majority of the BPH tissue site.

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: The invention concerns a container 1 with chambers 2 which each comprise at least one pair of electrodes including a first 4 and a second electrode 5 for the application of electric voltage for generating an electric field within one chamber 2. At least two first electrodes 4 of different chambers 3 are conductively coupled and at least one second electrode 5 of said chamber 2 is separately conductively connectable. The invention further concerns a method for manufacturing said container 1 as well as a device for electrically contacting at least one of said containers 1.

Abstract: This document discloses electroporation vessels, electrocompetent cells that have been aliquoted and frozen in electroporation vessels, and a number of other apparatuses, kits, and methods for electroporation. Some embodiments of electroporation vessels described herein may include a pair of opposing walls that are downwardly angled toward one another in a gap between two electrode surfaces. Further embodiments of devices and methods described herein may eliminate the need for an end user to transfer competent cells from a capped tube to electroporation cuvette, thereby saving time and producing less waste.

Abstract: Certain embodiments disclosed herein include, but are not limited to, at least one of compositions, methods, devices, systems, kits, or products regarding rejuvenation or preservation of germ cells or gametes. Certain embodiments disclosed herein include, but are not limited to, methods of modifying germ cells or gametes, or methods of administering modified germ cells or gametes to at least one biological tissue.

Abstract: In order to ensure consistently good maceration of biological material in an electroporation reactor, it is proposed to monitor the conductivity of the mixture therein and to detect any arcing which occurs therein. The results of such monitoring are used to modify the operating voltage of the electroporation reactor and/or the composition of the mixture located in the reactor channel.

Abstract: Disclosed is a circuit arrangement comprising at least one storage device for electrical charges to generate at least one voltage pulse by selectively discharging the storage device, and at least one control unit for controlling the discharge. A controller for monitoring the chronological progression of the voltage pulse is provided which controls at least one continuation of discharge after termination. Biomaterial is treated by using at least one electrical field generated by a first voltage pulse which is terminated once the value for an electrical parameter has exceeded or dropped below a preset limit. After the first voltage pulse has been terminated, it is continued by an additional voltage pulse.

Abstract: An electroporation cuvette is constructed with electroporation electrodes arranged in non-parallel relation to form a gap whose width varies with the location within the cuvette, plus a pair of positioning electrodes that are arranged to cause electrophoretic migration of biological cells within the cuvette according to cell size. Once the cells, suspended in a solution of the impregnant, are distributed in the cuvette by the positioning electrodes, electric field pulses are generated by the non-parallel electroporation electrodes. Because of their distribution in the cuvette, the various cells will experience voltage differentials across their widths that approach uniformity regardless of cell diameter, since the larger cells will be positioned at locations where the gap between the electrodes is greater and the smaller cells at locations where the gap is relatively small while the voltage drop across the entire gap is uniform along the length of the cell.

Abstract: A method and apparatus are provided for delivering an agent into a cell through the application of nanosecond pulse electric fields (“nsPEF's”). The method includes circuitry for delivery of an agent into a cell via known methods followed by the application of nanosecond pulse electric fields to said cell in order to facilitate entry of the agent into the nucleus of the cell. In a preferred embodiment, the present invention is directed to a method of enhancing gene expression in a cell comprising the application of nanosecond pulse electric fields to said cell. An apparatus for generating long and short pulses according to the present invention is also provided. The apparatus includes a pulse generator capable of producing a first pulse having a long duration and low voltage amplitude and a second pulse having a short duration and high voltage amplitude.

Abstract: Methods for material transfer into a cell are provided according to the present invention which include electroporation of the cell in the presence of the material, such as nucleic acids, and 2,3-butanedione monoxime; and lipofection with a complex of a lipid-based carrier, such as liposomes, and the material, in the presence of 2,3-butanedione monoxime.

Abstract: The present invention provides a method and apparatus for transferring an agent into a cell. The method includes the steps of providing an agent outside of a cell and generating a vapor bubble and a plasma discharge between an avalanche electrode and a conductive fluid surrounding the cell. The vapor bubble and plasma discharge generate a mechanical stress wave and an electric field, respectively. The combination of this mechanical stress wave and electric field results in permeabilization of the cell, which in turn results in transfer of the agent into the 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: The present invention provides for a system and method whereby aspirin and acetic acid help to permeabilize cell membranes to allow exogenous molecules to gain access to the cell interior. As such, the present invention provides a low cost drug and gene delivery tool that can be applied in combination with other molecular delivery methods.

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

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: The present invention comprises methods and systems for manipulation of media and particles, whether inert materials or biomaterials, such as cells in suspension cell culture. The methods and systems comprise use of an apparatus comprising a rotating chamber wherein the actions of the combined forces fluid flow force and centrifugal force form a fluidized bed within the rotating chamber.

Abstract: The present invention comprises methods and systems for manipulation of media and particles, whether inert materials or biomaterials, such as cells in suspension cell culture. The methods and systems comprise use of an apparatus comprising a rotating chamber wherein the actions of the combined forces of gravity, fluid flow force and centrifugal force form a fluidized bed within the rotating chamber.

Abstract: A method for introducing biologically active molecules into animal or human cells using electric current includes suspending the cells and dissolving the biologically active molecules in a buffer solution which has a buffer capacity of at least 20 mmol×l?1×pH?1 and an ionic strength of at least 200 mmol×l?1 at a change in the pH from pH 7 to pH 8 and at a temperature of 25° C. to form a suspension. The method further includes applying an electric voltage to the suspension so as to introduce the biologically active molecules into animal or human cells.

Abstract: The invention relates to a method of inducing pluripotency in a responsive mammalian cell, which comprises introducing into the cell an effective amount for initiating pluripotency within the cell of Oct4 protein or a functionally equivalent analogue, variant or fragment thereof. The invention also relates to a method of treatment and/or prophylaxis of a degenerative disease or injury in a mammal, which comprises removing from the mammal one or more responsive cells and culturing the cells in a suitable medium, introducing into the cells an effective amount of Oct4 protein or a functionally equivalent analogue, variant or fragment thereof and subsequently returning the cells to the patient. A further aspect of the invention relates to a method of treatment and/or prophylaxis of a degenerative disease or injury in a mammal, which comprises introducing into responsive cells of the patient an effective amount of Oct4 protein or a functionally equivalent analogue, variant or fragment thereof.

Abstract: Using electroporation, it is possible to activate the natural reprogramming potential of living Xenopus laevis oocytes and pass it on to donor cells placed with eggs in one electroporation chamber. We demonstrated that co-electroporation at 150 v/cm/25 ?F of mature oocytes with ˜105 cells/ml of suspension of various normal and cancerous human cell lines, such as bone marrow stromal cells, foreskin fibroblasts, pre-adipocytes, CD4+ T-lymphocytes, cheek cells, cervical carcinoma (HeLa) cells and breast adenocarcinoma (MCF-7) cells, reprograms donor cells into iPSc-like cells, which form colonies on irradiated MEF feeders. The iPSc-like cells generated by this study resemble human embryonic stem cells in colony morphology and expression of stem cell-associated transcription factors, including Oct3/4, Nanog, SOX-2, Rex-1, TRA-1-60 and SSEA-1. New method obviates the use of retroviral or lentiviral gene delivery vectors and other “non-parental” reprogramming approaches.

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 method for introducing biologically active molecules into animal or human cells using an electric current includes suspending the cells and dissolving the biologically active molecules in a buffer solution including HEPES and at least 10 mmol×1?1 magnesium ions (Mg2+), the buffer solution having a buffer capacity of at least 20 mmol×1?1×pH?1 at a change in the pH from pH 7 to pH 8 and at a temperature of 25° C., and an ionic strength of at least 200 mmol×1?1. An electric voltage is applied to the suspension.

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: The present invention provides a method and apparatus for transferring an agent into a cell. The method includes the steps of providing an agent outside of a cell and generating a vapor bubble and a plasma discharge between an avalanche electrode and a conductive fluid surrounding the cell. The vapor bubble and plasma discharge generate a mechanical stress wave and an electric field, respectively. The combination of this mechanical stress wave and electric field results in permeabilization of the cell, which in turn results in transfer of the agent into the cell.

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 present invention provides methods for de-differentiating somatic cells into stem-like cells without generating embryos or fetuses. More specifically, the present invention provides methods for effecting the de-differentiation of somatic cells to cells having stem cell characteristics, in particular pluripotency, by introducing RNA encoding factors inducing the de-differentiation of somatic cells into the somatic cells and culturing the somatic cells allowing the cells to de-differentiate.

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 provides an apparatus and a method for generating and applying an electric field according to a user configured arbitrary waveform pulsing train. A control module allows the user to input operation parameters and configure the arbitrary waveform pulsing train for the electroporation process. A micro-controller unit coupled to the control module controls an arbitrary waveform generator unit and a customized power module. During the electroporation process, the arbitrary waveform generator unit produces the arbitrary waveform pulsing train that is amplified by the customized power module. The customized power module is coupled to at least two electrodes, wherein the at least two electrodes will produce an electric field across a target media.

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 instant invention provides methods for the in vivo imaging of cells using one or more imaging modalities.

Abstract: The present invention is directed to methods of delivering macromolecules to a target cell or tissue by microwave irradiation. A target cell or tissue is exposed to one or more macromolecules to be delivered into the desired cell or tissue and irradiated with microwave radiation. The strength or power of the microwave radiation is such that the macromolecules are delivered into the target cell or tissue. Preferably, the strength of the microwave radiation does not significantly impact cell viability in a negative manner (e.g., apoptosis).

Abstract: A method of fractionating biomass, by permeability conditioning biomass suspended in a pH adjusted solution of at least one water-based polar solvent to form a conditioned biomass, intimately contacting the pH adjusted solution with at least one non-polar solvent, partitioning to obtain an non-polar solvent solution and a polar biomass solution, and recovering cell and cell derived products from the non-polar solvent solution and polar biomass solution. Products recovered from the above method. A method of operating a renewable and sustainable plant for growing and processing algae.

Abstract: The present invention provides a method and apparatus for transferring an agent into a cell. The method includes the steps of providing an agent outside of a cell and generating a vapor bubble and a plasma discharge between an avalanche electrode and a conductive fluid surrounding the cell. The vapor bubble and plasma discharge generate a mechanical stress wave and an electric field, respectively. The combination of this mechanical stress wave and electric field results in permeabilization of the cell, which in turn results in transfer of the agent into the cell.

Abstract: A method of treating biocells includes the steps of: a. providing biocells; b. applying at least one stressor to the biocells sufficient to cause nonlethal and reparable cell wall damage to the biocells, thereby putting the biocells in a catabolic state during which catabolic metabolic functions predominate over anabolic metabolic functions; and c. obtaining at least one product produced by the biocells during the catabolic state. In another embodiment, the method includes the steps of: a. providing biocells that are mammalian cells; b. applying at least one stressor to the biocells sufficient to cause nonlethal and reparable cell wall damage to the biocells, the reparable cell wall damage comprising openings that allow increased passage of materials through the cells walls; and c. inserting foreign DNA through the openings into the biocells.

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 configures 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: 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: 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 a method for treatment of benign prostatic hyperplasia are disclosed. The apparatus includes an applicator piece carrying a set of electrodes shaped and positioned to create a substantial electric field in the volume of hyperplasia and a pulse generator adapted for delivery of electrical pulses above the upper electroporation limit for the neoplastic cells. The amplitude, duration and number of the electrical pulses are generally selected to cause necrosis of a significant fraction of the volume of benign prostatic hyperplasia. The apparatus may include a high frequency system for heating the prostatic tissue and a cooling system for cooling the urethra. The combined action of heating and cooling may increase the temperature of the prostate cells to 45 degrees C. to 55 degrees C., while keeping the urinary tract at a temperature 15 degrees C. to 20 degrees C.