JPH0620397Y2 - Pulse generation circuit of cell fusion device - Google Patents

Description

TECHNICAL FIELD The present invention relates to a cell fusion device employing a cell electrofusion method used for improving the quality of plants and fungi by somatic cell hybrids, creating new species of bacteria, etc. The present invention relates to a pulse generation circuit (hereinafter referred to as a pulse generation circuit) of a cell fusion device that applies a voltage pattern required for fusion to a cell fusion electrode.

2. Description of the Related Art Recently, a cell electrofusion method in a cell fusion device is based on PEG.
Compared with (polyethylene glycol) method, calcium chloride method, etc., it is in the spotlight as a method that has high fusion efficiency, is simple, and has good stability.

A conventional example of a pulse generation circuit that is one of the main components of the cell fusion device is a sine wave generation circuit that outputs a sine wave for cell migration as a voltage, and a square wave that outputs a square wave pulse for cell fusion as a voltage. By alternately switching the pulse generation circuit, the sine wave generation circuit and the square wave pulse generation circuit, which are respectively connected to the cell fusion electrode, a sine wave and a square wave pulse are output to the cell fusion electrode in time division. It had a basic configuration with a relay.

However, in the above-mentioned conventional method, since the relay having the mechanical contact is included in the configuration that is mostly composed of electronic parts, the reliability of the pulse generation circuit, and in the case of the cell This has been an obstacle to improving the reliability of the fusion device.
Further, when a high-voltage square wave pulse or sine wave is required, there is a disadvantage that the entire circuit becomes large due to the breakdown voltage, and the price is greatly paid back accordingly.

The present invention has been made in view of the above circumstances, and an object of the present invention is to provide a pulse generation circuit improved so as to improve reliability as compared with the conventional one.

The pulse generation circuit according to the present invention is a circuit for repeatedly applying a time-divided bipolar wave for cell migration and a unipolar pulse for cell fusion to a cell fusion electrode. A unipolar pulse generation circuit that outputs a unipolar pulse according to a pulse control signal that gives the timing of pressurizing the polar pulse, and a time division that resets the wave control signal that controls the waveform of the bipolar wave according to the pulse control signal It is provided with a switching circuit, a bipolar wave generating circuit that outputs a bipolar wave according to a wave control signal, and a transformer that guides the bipolar wave and the unipolar pulse to the cell fusion electrode.

When appropriate, the time-divided bipolar wave and unipolar pulse are repeatedly output as a voltage.

Embodiment Hereinafter, an embodiment of the pulse generating circuit according to the present invention will be described with reference to the drawings. FIG. 1 is an electrical connection diagram of a pulse generating circuit, and FIG. 2 is a timing chart showing a pressure waveform of a cell fusion electrode and main signals.

Although not shown in the figure, inside the cell fusion device that employs the cell electrofusion method, there is a chamber in which the cell suspension is immersed, and a cell fusion electrode 50 that gives an electric field to the suspended cells is provided therein. The provided cell fusion device is a device that fuses cells by giving an electric field of a predetermined pattern to clouded cells, and the pulse generation circuit is a central control unit (not shown) of this device. It is a circuit that creates a cell fusion pattern, that is, a voltage pattern that pressurizes the cell fusion electrode 50, based on a command from. This voltage pattern has a form in which a time-divided sine wave (bipolar wave) for cell migration and a square wave pulse (unipolar pulse) for cell fusion are alternately repeated (see FIG. 2). ). The command from the central control unit is introduced into the pulse generation circuit as a pulse control signal 62 that gives the timing of pressurizing the square wave pulse and a wave control signal 63 that controls the waveform of the sine wave. However, the wave control signal 63 is a signal that continuously changes regardless of the timing given by the pulse control signal 62. Hereinafter, the configuration of the unipolar pulse generation circuit 10 that outputs a square wave pulse according to the pulse control signal 62 will be described.

The unipolar pulse generation circuit 10 rectifies the AC voltage introduced from the power supply line 61 with the diode 11 and the smoothing capacitor 12 and switches the rectified DC voltage with the FET 13 whose pulse control signal 62 is connected to the gate. It has become a structure. A primary circuit 41 of a transformer 40 described later is connected to the output terminal of this circuit. The pulse control signal 62 introduced via the current buffer 621 is also guided to the time division switching circuit 20 described later. Further, the FET 13 is provided with a protection diode 14. Next, the bipolar wave generation circuit 30 that outputs a sine wave according to the wave control signal 63 will be described.

The bipolar wave generating circuit 30 is an inverter circuit configured to generate a Sany wave by switching the DC voltage of the power source 36 to both polarities by the FETs 32 and 33 which are designed to operate alternately. Gate has a current buffer
The wave control signal 63 via 631 is introduced, and FET32
An inverted wave control signal 631 obtained by inverting the wave control signal 63 by the inverter 31 is introduced into the. The primary circuit 42 of the transformer 40 is connected to the output terminal of this circuit. Protective diodes 34 and 35 are provided between the anodes and drains of the FETs 32 and 33, respectively. Hereinafter, the time division switching circuit 20 that operates so that the sine wave and the square wave pulse obtained as described above are alternately output will be described.

The time division switching circuit 20 is two inversion circuits that invert the pulse control signal 62, and the wave control signal 63 and the inverted wave control signal are generated at the timing when the pulse control signal 62 becomes active.
It is configured to forcibly reset the 631.
This is because when the square wave pulse is output from the unipolar pulse generation circuit 10, the FET of the bipolar wave generation circuit 30 is
This means that both 32 and 33 are turned off.

By the way, as described above, the transformer 40 to which the unipolar pulse generating circuit 10 and the bipolar wave generating circuit 30 are connected is a step-up pulse transformer having two inputs and one output, and the square wave pulse and the sine wave are overlapped. The pressure is also increased and the cell fusion electrode 50 is pressed. The operation of the pulse generating circuit configured as described above will be briefly described below with reference to FIG. 2 (however, the time relationship in FIG. 2 is not accurate).

In the period T1 when the pulse control signal 62 is at H level,
The FET 13 is turned on, while both the FETs 32 and 33 are turned off because the wave control signal 62 and the inverted wave control signal 631 are reset by the time division switching circuit 20. That is, only the unipolar pulse generation circuit 10 is operating during this period, and the cell fusion electrode 50 is pressurized with the square wave pulse of the voltage E ₂ boosted by the transformer 40.

Further, during the period T2 when the pulse control signal 62 is at L level, the FET 13 is switched to the off state,
Both 32 and 33 are released from the reset in the period T1 and operate according to the wave control signal 63. That is, only the bipolar wave control circuit 30 is operating during this period, and the sine wave of the amplitude voltage E ₁ boosted by the pulse transformer 40 is applied to the cell fusion electrode 50.
Note that since the operation is exactly the same after the period T3, description thereof is omitted.

In the case of the present embodiment, since the transformer 40 is configured to boost the square wave pulse and the sine wave, even if it is necessary to apply a high voltage to the cell fusion electrode 50, the unipolar pulse Generation circuit 10 and bipolar wave generation circuit
There is a merit that 30 can be substantially simplified. However, it goes without saying that the transformer 40 may be a transformer having a simple winding ratio of 1: 1.

The pulse generating circuit is not limited to the above-mentioned embodiment, and a transformer with a tap may be adopted to switch the input appropriately. In this case, the applied voltage of the cell fusion electrode 50 can be changed.

Regarding the voltage waveform applied to the cell fusion electrode 50,
Any waveform may be used as long as it is a repeating waveform of a bipolar wave and a unipolar pulse.

Effect of the Invention According to such a pulse generating circuit, since the configuration does not include a relay, it is possible to significantly improve the reliability of the circuit, that is, the reliability of the cell fusion device, as compared with the related art. Further, due to the fact that the relay is not included, there are various effects that the circuit configuration can be simplified and downsized.

[Brief description of drawings]

FIG. 1 is an electrical connection diagram of an embodiment of a pulse generating circuit according to the present invention, and FIG. 2 is a timing chart showing a pressurizing waveform of a cell fusion electrode and main signals. 10 …… Monopolar pulse generation circuit 20 …… Time division switching circuit 30 …… Bipolar wave generation circuit 40 …… Transformer 50 …… Cell fusion electrode 62 …… Pulse control signal 63 …… Wave control signal

Claims (1)

[Scope of utility model registration request] 1. A circuit for repeatedly applying a time-divided bipolar wave for cell migration and a monopolar pulse for cell fusion to a cell fusion electrode, the pulse giving a timing of pressurizing the monopolar pulse. A unipolar pulse generation circuit that outputs the unipolar pulse according to a control signal; a time division switching circuit that resets a wave control signal that controls the waveform of the bipolar wave according to the pulse control signal; and the wave control A pulse for a cell fusion device, comprising: a bipolar wave generation circuit that outputs the bipolar wave according to a signal; and a transformer that guides the bipolar wave and the monopolar pulse to the cell fusion electrode. Generator circuit.