JP6518971B2 - Electroporator power supply - Google Patents

Description

  The present invention relates to an electroporator power source, an electroporator including the same, and an electroporation method using the same.

  Various electroporators are known as an apparatus for performing an electroporation to open micropores in cell membranes by electrical stimulation and introduce macromolecules such as DNA into cells. As such an electroporator, for example, in vitro electroporation is performed on culture cells suspended in an appropriate buffer solution, culture cells in a state of being attached to a culture vessel, and the like. And those for performing in vivo electroporation for living tissue etc. are commercially available, and the administration device for DNA vaccine is one of the electroporators. is there.

  Early electroporators loaded high-voltage square-wave pulses with a constant voltage multiple times at the desired pulse length, pulse interval, and number of pulses, but to increase electroporation efficiency, high-voltage After applying a pulse for a relatively short time (on the order of 10 microseconds), a dual pulse type electroporation was subsequently developed to load a low voltage pulse for a longer time (on the order of 10 milliseconds) (non-patented Literature 1). In this method, the first high voltage pulse contributes to the formation of fine pores in the cell membrane (poration pulse; hereinafter abbreviated as Pp), and the subsequent low voltage pulse contributes to the movement into the inside of the details such as DNA. It is considered to be (driving pulse, hereinafter Pd). Although the electroporation described in the above-mentioned Non-Patent Document 1 loads one pulse both in the poration pulse and the driving pulse, after loading one poration pulse (Pp), A device (CUY21EX, BEX Co., Ltd.) that can load a square wave pulse at a constant voltage multiple times (Figure 1) or an exponential decay wave multiple times (Figure 3) as a driving pulse (Pd) Are also commercially available.

  The electroporator before such a dual pulse type electroporation apparatus was developed was only equipped with a single power supply for outputting a square wave pulse of a constant voltage, but a dual pulse type This problem has been solved by providing two constant voltage sources, since it is necessary to output two types of pulses having substantially different voltages with a short time interval between them.

Biophys. J., 1992, Vol. 63, p.

  An object of the present invention is to provide an electroporation power source that is implemented by combining two or more types of pulses, and to provide an electroporation power source capable of achieving a higher introduction rate.

The present invention
[1] It has two or more independent DC power supply units that can be used in conjunction with each other,
At least one of the DC power supply units includes a constant current source capable of outputting one or more pulses of unipolarity at a predetermined current value;
[2] As the DC power supply unit,
(A) DC power supply unit (Pp / P) for outputting a poration pulse (Pp);
(B) A power supply for electroporator of [1], including a DC power supply unit (Pd / P) for outputting a driving pulse (Pd);
[3] The DC power supply unit (Pd / P)
(B2) A constant current source (Pd / a) capable of outputting one or a plurality of unipolar pulses at a predetermined current value as a driving pulse (Pd)
[2] power source for electroporator including;
[4] The DC power supply unit (Pd / P) further includes:
(B1) A constant voltage source (Pd / p) capable of outputting one or a plurality of unipolar pulses at a predetermined voltage value as a driving pulse (Pd)
[3] power sources for electroporators, including:
[5] As the DC power supply unit (Pd / P),
(B ⁺ ) DC power supply unit (Pd + / P) for outputting positive polarity driving pulse (Pd +);
(B ⁻ ) A power supply for electroporator of [2], including a DC power supply unit (Pd − / P) for outputting a negative driving pulse (Pd ⁻ );
[6] As the constant current source (Pd / a),
(B2 ⁺⁾ as a positive polarity driving pulse (Pd +), a constant current source for one or more pulses of positive polarity can be output at a predetermined current value (Pd + / a) and (B2 ^-) negative driving pulse ( A power source for electroporator of [3], including a constant current source (Pd− / a) capable of outputting one or a plurality of pulses of negative polarity at a predetermined current value as Pd−);
[7] As the constant voltage source (Pd / p),
(B1 ⁺⁾ as a positive polarity driving pulse (Pd +), constant voltage source to one or more pulses of positive polarity can be output at a predetermined voltage value (Pd + / p) and (B1 ^-) negative driving pulse ( The electroporate of [4] comprising Pd-), a constant voltage source (Pd- / p) capable of outputting an exponential decay wave consisting of one or more pulses of negative polarity at a predetermined voltage value. Power supply for
[8] The DC power supply unit (Pp / P)
(A1) A constant voltage source (Pp / p) capable of outputting one or a plurality of unipolar pulses at a predetermined voltage value as a poration pulse (Pp), and (A2) as a poration pulse (Pp) , A constant current source (Pp / a) capable of outputting one or more pulses of unipolarity at a predetermined current value
A power source for an electroporator according to any one of [2] to [7], comprising at least one selected from the group consisting of
[9] An electroporator comprising the electroporator power source according to any one of [1] to [8] and an electrode for electroporation;
[10] An electroporation method characterized by using the electroporator of [9];
About.

In the present specification, “constant voltage source” means not only a power supply capable of outputting a plurality of pulses at a constant voltage, but also, for example, outputs a plurality of pulses at a predetermined voltage preset by an initial setting voltage and a reduction rate. It also means the power that you can.
Further, in the present specification, "constant current source" means not only a power supply capable of outputting a plurality of pulses at a constant current value, but also a power supply capable of outputting a plurality of pulses at a predetermined current value set in advance.

According to the power supply for electroporator of the present invention, since it is possible to output a plurality of pulses at a predetermined current value set in advance, it is possible to carry out the electroporation performed under the condition that the resistance value is expected to fluctuate during the load. Even if it is, high introduction efficiency can be achieved.
Further, according to a preferred embodiment of the present invention, a DC power supply unit (Pd + / P) for outputting a positive polarity driving pulse (Pd +) and a DC power supply unit for outputting a negative polarity driving pulse (Pd−) According to the power supply for the electroporator provided with (Pd− / P), it is possible to output the driving pulse of the pattern (for example, FIGS. 4 to 9) which can not be implemented by the electroporator so far.

In the dual pulse type electroporation (hereinafter the same in FIGS. 2 to 9) loading a driving pulse (Pd) following a poration pulse (Pp), a square wave pulse of positive polarity as a driving pulse (Pd) Is a schematic waveform diagram in the case of outputting a plurality of times. It is a typical waveform figure in the case of outputting an exponential decay wave which consists of a plurality of positive polarity pulses (each pulse is itself a square) as a driving pulse (Pd) in the same electroporation. In the same electroporation, it is a typical waveform diagram in the case of outputting exponential decay wave (each pulse itself is also exponential decay (exponential)) which consists of a plurality of positive polarity pulses as a driving pulse (Pd). . In the same electroporation, after outputting a square wave pulse of positive polarity multiple times as a driving pulse (Pd), it is a typical waveform diagram in the case of outputting a rectangular wave pulse of negative polarity multiple times more. FIG. 7 is a schematic waveform diagram in the case of alternately outputting positive square wave pulses and negative square wave pulses alternately as a driving pulse (Pd) in the same electroporation. In the same electroporation, as the driving pulse (Pd), a positive polarity exponential decay wave (each pulse is itself a square) is output a plurality of times, and then a negative polarity exponential decay wave (each The pulse itself is a schematic waveform diagram in the case of outputting a square multiple times. In the same electroporation, as a driving pulse (Pd), an exponentially decaying wave of positive polarity (each pulse is itself a square) and an exponentially decaying wave of negative polarity (each pulse is a square (square) ) Is a schematic waveform diagram in the case of alternately outputting one by one. In the same electroporation, as the driving pulse (Pd), a positive polarity exponential decay wave (each pulse itself is also output exponential decay multiple times) is output a plurality of times, and then negative polarity exponential decay It is a typical waveform figure in the case of outputting a wave (each pulse itself is also exponential decay (exponential)) multiple times. In the same electroporation, as a driving pulse (Pd), an exponentially decaying wave of positive polarity (each pulse itself is also exponentially decaying) and an exponentially decaying wave of negative polarity (each pulse is itself too FIG. 6 is a schematic waveform diagram in the case of alternately outputting exponential decay (exponential) once. It is a block diagram which shows typically the structure of the one aspect | mode of the power supply for electroporators of this invention. FIG. 11 is a block diagram showing a constant voltage source (Pp / p) functioning as a DC power supply unit (Pp / P) for outputting a poration pulse (Pp) in a block diagram shown in FIG. 10 by a thick line. FIG. 11 is a block diagram showing, in thick lines, constant voltage sources (Pd + / p) constituting a DC power supply unit (Pd + / P) for positive polarity driving pulse (Pd +) output in the block diagram shown in FIG. 10; FIG. 11 is a block diagram showing, in thick lines, constant current sources (Pd + / a) constituting a DC power supply unit (Pd + / P) for positive polarity driving pulse (Pd +) output in the block diagram shown in FIG. 10;

The power supply for electroporator of the present invention includes at least two or more (preferably three or more) independent DC power supply units that can be used singly or in conjunction. Such a combination of DC power supply units is not particularly limited as long as it is a combination usable for electroporation, but, for example, a combination of a high voltage output power supply and a low voltage output power supply is mentioned In particular, a combination of a DC power supply unit (Pp / P) for outputting a poration pulse (Pp) and a DC power supply unit (Pd / P) for outputting a driving pulse (Pd) is cited. be able to.
The power supply for electroporator of the present invention can also be provided with an alternating current power supply unit in addition to the direct current power supply unit.

  For example, a constant voltage source capable of outputting a square wave pulse of a constant voltage a plurality of times, or an index consisting of one or more pulses of single polarity as a DC power supply unit used in a conventional electroporator power supply A constant voltage source capable of outputting a functional attenuation wave at a predetermined voltage value may be mentioned, but in the electroporator power source according to the present invention, at least one of the DC power source units has one or more pulses of unipolarity. It includes a constant current source capable of outputting at a predetermined current value.

  For example, the power supply for electroporator of the present invention includes a DC power supply unit (Pp / P) for outputting a poration pulse (Pp) and a DC power supply unit (Pd / P) for outputting a driving pulse (Pd). And one of the DC power supply unit (Pp / P) and the DC power supply unit (Pd / P) may include the constant current source, or the DC power supply unit (Pp / P) and the DC power supply unit. Both (Pd / P) can also include the constant current source.

  The DC power supply unit (Pp / P) for outputting a poration pulse (Pp) that can be used in the present invention has, for example, one or more pulses of unipolarity as a poration pulse (Pp) and a predetermined voltage Value can be output as a constant voltage source (Pp / p) or as a poration pulse (Pp), a constant current source (Pp) capable of outputting one or a plurality of unipolar pulses at a predetermined current value / A), and either the constant voltage source (Pp / p) or the constant current source (Pp / a) or both may be included.

  Similarly, a DC power supply unit (Pd / P) for outputting a driving pulse (Pd) that can be used in the present invention is, for example, of outputting one or more pulses of unipolarity at a predetermined voltage value. It is possible to include a constant current source (Pd / a) capable of outputting one or a plurality of unipolar pulses at a predetermined current value as a constant voltage source (Pd / p) that can be used or a driving pulse (Pd). It is possible to provide either the constant voltage source (Pd / p) or the constant current source (Pd / a) or both.

  In ordinary dual pulse type electroporation, the number of loadings of driving pulses is often one. Therefore, in such an application, the constant voltage source (Pp / p) is considered in consideration of the easiness of construction and cost. DC power supply unit for output for poration pulse (Pp) consisting only of (Pp), DC power supply for driving pulse (Pd) output consisting of the constant voltage source (Pd / p) and constant current source (Pd / a) A combination with part (Pd / P) is preferred.

  The electroporator power source according to the present invention has two types for outputting driving pulses (Pd +) of different polarities in various combinations as a DC power source (Pd / P) for outputting driving pulses (Pd). DC power supply unit, that is, a DC power supply unit (Pd + / P) for outputting a positive polarity driving pulse (Pd +), and a DC power supply unit (Pd− for outputting a negative polarity driving pulse (Pd−) And (P) can be included.

  A DC power supply unit (Pd / P) for outputting a driving pulse (Pd) that can be used in the present invention; and a DC power supply unit (Pd + / P) for outputting a driving pulse (Pd +) of positive polarity; When the DC power supply unit (Pd− / P) for outputting the negative polarity driving pulse (Pd−) is included, each DC power supply unit (Pd + / P) and (Pd− / P) is as described above. As a constant voltage source (Pd / p) capable of outputting one or more pulses of unipolarity at a predetermined voltage value and / or one or more pulses of unipolarity as a driving pulse (Pd) A constant current source (Pd / a) capable of outputting at a predetermined current value can be included.

Specifically, the DC power supply unit (Pd + / P) for outputting the positive polarity driving pulse (Pd +) has a predetermined voltage value of one or a plurality of positive polarity pulses as the positive polarity driving pulse (Pd +). And / or a constant current source capable of outputting one or more pulses of positive polarity at a predetermined current value as the constant voltage source (Pd + / p) and / or the positive polarity driving pulse (Pd +) (Pd + / a) can be included.
Further, the DC power supply unit (Pd− / P) for outputting the negative driving pulse (Pd−) is a negative driving pulse (Pd−), and one or more negative polarity pulses have a predetermined voltage value. Constant voltage source (Pd − / p) and / or negative polarity driving pulse (Pd −) that can output at a predetermined current value. And a current source (Pd− / a).

Hereinafter, the present invention will be further described based on FIGS. 10 to 13 by taking one aspect of the power supply for electroporator of the present invention shown in FIG. 10 as an example. One embodiment of the present invention shown in FIG.
A DC power supply unit (Pp / P) for outputting a poration pulse (Pp);
A DC power supply unit (Pd + / P) for outputting a positive polarity driving pulse (Pd +);
DC power supply unit (Pd− / P) for outputting negative polarity driving pulse (Pd−);
As the DC power supply unit (Pp / P) for output of the above-mentioned poration pulse (Pp), it includes a constant voltage source (Pp / p) capable of outputting one or a plurality of unipolar pulses at a predetermined voltage value;
And a constant voltage source (Pd + / p) capable of outputting one or a plurality of pulses of positive polarity at a predetermined voltage value as the DC power supply unit (Pd + / P) for outputting the positive polarity driving pulse (Pd +). A constant current source (Pd + / a) capable of outputting one or more pulses of positive polarity at a predetermined current value;
A constant voltage source (Pd− / p) capable of outputting one or a plurality of negative polarity pulses at a predetermined voltage value as the DC power supply unit (Pd− / P) for the negative polarity driving pulse (Pd−) output And a constant current source (Pd− / a) capable of outputting one or more pulses of negative polarity at a predetermined current value.

  11 to 13 show one of each DC power supply unit by a thick line in the embodiment shown in FIG. FIG. 11 shows only a constant voltage source (Pp / p) functioning as a DC power supply unit (Pp / P) for outputting a poration pulse (Pp) in bold lines, and FIG. 12 shows a positive polarity driving pulse (Pd + Only the constant voltage source (Pd + / p) constituting the output DC power source (Pd + / P) is indicated by a thick line, and FIG. 13 shows the DC power source (Pd + / P) for positive polarity driving pulse (Pd +) output. Only the constant current source (Pd + / a) which comprises is shown by a thick line.

  In the DC power supply unit (Pp / P) for poration pulse (Pp) output shown in FIG. 11, by switching the switch (Pp output), the Pp power is stored in the capacitor (Pp capacity) and input to the Pp output switch According to the Pp drive signal, one or more unipolar pulses can be output from the output terminal at a predetermined voltage value.

  In the DC power supply unit (Pd + / P) for outputting positive polarity driving pulse (Pd +) in FIG. 10, one or more pulses of positive polarity shown in FIG. 12 are output at a predetermined voltage value by switch operation. Can be switched between a constant voltage source (Pd + / p) capable of generating a constant current source (Pd + / a) capable of outputting one or a plurality of pulses of positive polarity at a predetermined current value as shown in FIG. . Specifically, by switching the switch to the capacitor mode, it is possible to cause the DC power supply unit (Pd + / P) for positive polarity driving pulse (Pd +) output to function as a constant voltage source (Pd + / p). Further, by switching the switch to the amplifier mode, it is possible to cause the DC power supply unit (Pd + / P) for positive polarity driving pulse (Pd +) output to function as a constant current source (Pd + / a).

  In the constant voltage source (Pd + / p) shown in FIG. 12, the switch is switched to store the capacitor (Pd capacity) from the Pd + power supply and at a predetermined voltage value according to the Pd + drive signal input to the Pd + output switch. One or more unipolar pulses can be output from the output terminal.

On the other hand, in the constant current source (Pd + / a) shown in FIG. 13, the applied voltage of the Pd + power supply is directly output from the output terminal according to the Pd + drive signal input to the Pd + output switch. At that time, the current value and the voltage value are detected, and a feedback can be made to output a predetermined current value from the output terminal.
In the power supply for electroporator of the present invention, this feedback can be implemented, for example, using a semiconductor ± switch amplifier.

The above-described DC power supply unit for negative driving pulse (Pd−) output (Pd− / P) in FIG. 10 has a positive polarity driving pulse (Pd +) output DC power supply unit (Pd + / P) except that the polarity is opposite. It has the same configuration as P), and operates in the same manner as the DC power supply (Pd + / P) for positive polarity driving pulse (Pd +) output.
That is, by switching the switch to the capacitor mode, it is possible to cause the DC power supply unit (Pd− / P) for negative driving pulse (Pd−) output to function as a constant voltage source (Pd− / p). Further, by switching the switch to the amplifier mode, it is possible to cause the DC power supply unit (Pd− / P) for negative polarity driving pulse (Pd−) output to function as a constant current source (Pd− / a).
In addition, the constant voltage source (Pd− / p) and the constant current source (Pd− / a) are also constant voltage source (Pd + / p) and constant current source (Pd + / a), respectively, except that the polarities are opposite to each other. It works in the same way as).

FIGS. 1 to 9 show waveform patterns that can be output by the power supply for electroporator of the present invention in dual pulse type electroporation in which a driving pulse (Pd) is loaded following a poration pulse (Pp).
FIG. 1 shows a square wave pulse of positive polarity as a driving pulse (Pd) following a poration pulse (Pp) outputted by a constant voltage source (Pp / p) (hereinafter the same in FIGS. 2 to 7). It is output multiple times. When the positive rectangular wave pulse is output by a constant current source (Pd + / a), the rectangular wave pulse can be loaded at a constant current value. Moreover, when it outputs with a constant voltage source (Pd + / p), the square wave pulse of fixed voltage can be loaded.

In FIG. 2, following the same poration pulse (Pp), an exponential decay wave consisting of a plurality of positive polarity pulses (each pulse is itself a square) is output as a driving pulse (Pd) a plurality of times It is a thing. The exponentially decaying wave may be loaded at a predetermined current value preset by an initial setting current and a reduction rate by outputting the positive polarity exponentially decaying wave by the constant current source (Pd + / a). it can.
FIG. 3 shows, as the driving pulse (Pd), a plurality of exponential decay waves (each pulse itself is also exponential decay) as the driving pulse (Pd) following the same poration pulse (Pp) It will be output once. By outputting the positive polarity exponentially decaying wave by the constant voltage source (Pd + / p), the exponentially decaying wave can be loaded at a predetermined voltage preset by the initial setting voltage and the decrease rate etc. .

  1 to 3 output a square wave pulse or exponentially decaying wave of positive polarity, but in place of the constant voltage source (Pd + / p) or the constant current source (Pd + / a), By using a constant voltage source (Pd− / p) or a constant current source (Pd− / a), a square wave pulse or exponentially decaying wave of negative polarity (ie, the opposite polarity to the poration pulse (Pp)) is obtained. It can be output.

FIG. 4 shows that a rectangular pulse of positive polarity is output a plurality of times as a driving pulse (Pd) following the poration pulse (Pp), and then a rectangular pulse of further negative polarity is output a plurality of times . Load a rectangular wave pulse at a constant current value by outputting a positive rectangular wave pulse by a constant current source (Pd + / a) and then outputting a negative rectangular wave pulse by a constant current source (Pd− / a) be able to. Moreover, when it outputs with a constant voltage source (Pd + / p) and a constant voltage source (Pd- / p) succeedingly, the square wave pulse of a fixed voltage can be loaded.
In FIG. 5, following the same poration pulse (Pp), as a driving pulse (Pd), a positive rectangular wave pulse and a negative rectangular wave pulse are alternately output once at a time. Loading a rectangular wave pulse with a constant current value by outputting a positive rectangular wave pulse by a constant current source (Pd + / a) and outputting a negative rectangular wave pulse by a constant current source (Pd− / a) it can. Moreover, when it outputs by a constant voltage source (Pd + / p) and a constant voltage source (Pd- / p), the square wave pulse of a fixed voltage can be loaded. Although FIG. 5 alternately outputs a positive pulse and a negative pulse one time each, it is also possible to alternately output a plurality of positive pulses and the same number of negative pulses.

FIG. 6 shows that a positive polarity exponential decay wave (each pulse itself is a square) is output several times as a driving pulse (Pd) following the same poration pulse (Pp), and then the negative electrode is further generated. An exponentially decaying wave of sex (each pulse is itself a square) is output multiple times. By setting the positive polarity exponential decay wave by the constant current source (Pd + / a) and then the negative polarity exponential decay wave by the constant current source (Pd− / a), the preset current and the reduction rate etc. The exponentially decaying wave can be loaded at a predetermined current value set in advance.
FIG. 7 shows, as the driving pulse (Pd), an exponentially decaying wave of positive polarity (each pulse itself is a square) and an exponentially decaying wave of negative polarity following the same poration pulse (Pp) (Each pulse itself alternately outputs square once). By outputting a positive power exponentially decaying wave with a constant current source (Pd + / a) and a negative power exponentially decaying wave with a constant current source (Pd− / a), the preset current and reduction rate etc. Exponential decay waves can be loaded at a set predetermined current value. Although FIG. 7 alternately outputs the positive polarity pulse and the negative polarity pulse once each, it is also possible to alternately output a plurality of positive polarity pulses and the same number of negative polarity pulses.

FIG. 8 shows that a positive polarity exponential decay wave (each pulse itself is also output exponential decay multiple times) as a driving pulse (Pd) following the same poration pulse (Pp) Furthermore, an exponentially decaying wave of negative polarity (each pulse itself is also exponentially decayed) is output a plurality of times. By setting the positive polarity exponential decay wave by the constant voltage source (Pd + / p) and then the negative polarity exponential decay wave by the constant voltage source (Pd− / p), the initial setting voltage and the reduction rate etc. The exponential decay wave can be loaded at a predetermined voltage set in advance.
FIG. 9 shows, as the driving pulse (Pd), an exponential decay wave of positive polarity (each pulse itself is also exponential decay (exponential)) and an exponential function of negative polarity following the same poration pulse (Pp) Dynamic attenuation waves (each pulse itself is also exponentially decayed) once alternately. By outputting a positive polarity exponential decay wave with a constant voltage source (Pd + / p) and a negative polarity exponential decay wave with a constant voltage source (Pd− / p), the preset voltage and reduction rate etc. Exponential decay waves can be loaded at a set predetermined voltage. Although FIG. 9 alternately outputs a positive pulse and a negative pulse one time each, it is also possible to alternately output a plurality of positive pulses and the same number of negative pulses.

  Although the specification of the power supply for electroporator of this invention can be determined suitably, the example is described below. The DC power supply unit for outputting positive polarity driving pulse (Pd +) (Pd + / P) and the DC power supply unit for outputting negative polarity driving pulse (Pd−) (Pd− / P) are opposite in polarity except for the polarity. Since the specifications are the same, the description of the latter is omitted.

DC power supply for poration pulse (Pp) output (Pp / P) [constant voltage source (Pp / p)]
・ Output voltage: 0 to 400 V, setting unit: 1 V steps ・ Capacitor: 1410 μF
· Handling output current: 0.01 to 10 A
・ Number of pulses: 1
Pulse width: 10 microseconds to 99.9 milliseconds Interval between poration pulse and driving pulse: 50 microseconds to 99.9 milliseconds

Constant voltage source (Pd + / p) in DC power supply (Pd + / P) for positive polarity driving pulse (Pd +) output
・ Output voltage: 0 to 300 V, setting unit: 1 V
Capacitor: 22 μF, 55 μF, 139 μF, 349 μF, 876 μF, 2200 μF
· Handling output current: 0 to 10 A
-Number of pulses: 1 to 1000
Pulse width: 50 microseconds to 1000 milliseconds Pulse interval: 50 microseconds to 1000 milliseconds

Constant current source (Pd + / a) in DC power supply (Pd + / P) for positive polarity driving pulse (Pd +) output
・ Output voltage: 1 to 200V, setting unit: 1V
・ Output current: 1 to 1000 mA, setting unit: 1 mA
-Number of pulses: 1 to 1000
Pulse width: 50 microseconds to 1000 milliseconds Pulse interval: 50 microseconds to 1000 milliseconds

  The electroporator power source according to the present invention does not damage cells from the AC power source before the pulse output (for example, the output of the poration pulse (Pp) and the driving pulse (Pd)) originally intended. It is possible to provide a means for measuring the resistance between the electroporation electrodes by passing a relatively weak constant current. In the power supply for electroporator of the present invention, such resistance measurement means and each of the DC power supply units described above can be implemented as a series of continuous operations.

  In addition, the power source for the electroporator of the present invention can be provided with means for preventing sudden overpowering for safety. Such prevention means may either stop output in hardware or limit output in software. As software over-output prevention means, for example, the resistance of the load is measured in advance, and the set output current is set by setting the output limit voltage, or the output limit current is set for the set output voltage By setting, it is possible to prevent sudden overpowering.

  In addition, in the power supply for electroporator of the present invention, in the case of outputting exponential decay waves in a constant current source (for example, Pd + / a, Pd− / a), according to the attenuation rate, as desired, The power supply voltage of the power supply (e.g., Pd + power supply, Pd − power supply) can be lowered in advance. Thereby, energy loss (heat generation) of the amplifier can be reduced.

  Further, in the power supply for electroporator of the present invention, even if a large output is required by setting the power supply voltage slightly higher (about 10 to 20%) than the set voltage, if desired, The arrival time to the set voltage can be shortened.

The electroporator power source of the present invention can be used in combination with appropriate electroporation electrodes to carry out various electroporations.
Examples of electroporation that can be performed using the power supply for electroporator of the present invention include, for example, in vivo electroporation (for example, in utero electroporation, ex utero electroporation, and the like). ovo electroporation), ex ovo electroporation, ex vivo electroporation, in vitro electroporation (eg, electroporation in suspension, electro Can be mentioned.

  The power source for the electroporator of the present invention is used in electroporation in which micropores are formed in the cell membrane by electrical stimulation and a polymer such as DNA is introduced into cells by using it in combination with the electrode for electroporation. be able to.

Claims (3)

Two or more independent DC power supply units that can be used in conjunction with each other,
And an output terminal,
The two or more independent DC power supply units are
One is a poration pulse DC power supply for outputting a poration pulse (Pp) (Pp / P) ,
Another one is a driving pulse DC power supply for outputting a driving pulse (Pd) (Pd / P) ,
A constant current in which at least one of the poration pulse DC power supply unit (Pp / P) and the driving pulse DC power supply unit (Pd / P) can output one or more pulses of a single polarity at a predetermined current value Including the source,
First, the poration pulse (Pp) is output from the output terminal, and then the driving pulse (Pd) is output.
The absolute value of the voltage of the poration pulse (Pp) is greater than the absolute value of the voltage of the driving pulse (Pd),
A power supply for electroporator, wherein the constant current source can output a predetermined current value from the output terminal by detecting and feeding back a current value output from the output terminal. A power source for an electroporator according to claim 1 ;
An electrode for electroporation;
Including the electroporator. An electroporation method comprising using the electroporator according to claim 2 .

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