WO2018195682A1 - Blot transfer device, blot transfer system, and control method - Google Patents

Abstract

A blot transfer device (10), blot transfer system, and control method. The blot transfer device (10) comprises a control unit (11) and one or more blot transfer units (130). The control unit (11) is configured to apply to a first planar electrode layer and a second planar electrode layer of the one or more blot transfer units (130) a monopolar electrical signal, the monopolar electrical signal varying according to a predetermined amplitude period, wherein a ratio of an instantaneous power corresponding to the peak of the predetermined amplitude to an instantaneous power corresponding to the lowest point of the predetermined amplitude is greater than or equal to 2. The blot transfer device has superior blotting transfer efficiency, and ensures excellent transfer efficiency and quality.

Description

Biotransfer device, biotransfer system and control method Technical field

The invention belongs to the technical field of biochemical analysis and relates to the transfer of biological macromolecules, in particular to a biotransfer device using a unipolar electrical signal with varying fluctuations, a biotransfer system and a control method thereof.

Background technique

A biotransfer device (Blot Transfer Device) is a method for transferring a biomacromolecule (for example, a protein) in a gel layer containing a biomacromolecule onto a carrier film by using an electrophoresis principle. Therefore, the biotransfer device generally includes a control unit. It is used to generate a corresponding electrical signal applied to the gel layer to control the above-described electrophoresis-based transfer process, the electrical signal being applied specifically to the electrodes of the transfer unit comprising the gel layer and the carrier film.

Taking Western Blot Transfer as an example, the molecular size of proteins during transfer is usually inconsistent. For different sizes of proteins, the transfer rate under the same electrical signal is different, especially for relative Larger proteins have a relatively low rate of transfer. At present, it is generally considered that the transfer efficiency and the transfer quality in the biotransfer device are proportional to the voltage applied to the transfer unit, and therefore, the transfer efficiency is improved and the good is obtained by increasing the voltage applied to the transfer unit. Transfer quality. However, such a method of improving the voltage applied to the transfer unit or extending the transfer time blindly has a problem that the transfer unit is easily overheated and the transfer quality is poor, and the transfer efficiency is not necessarily ensured.

In the solution for improving the transfer efficiency of the Western blotting device disclosed in the patent No. US8721860B2, entitled "Protein Multi-blotting Method and Device", one of the upper and lower electrode layers of the transfer unit terminal of the protein transfer device is In the multiple composite electrode layer, different electrical signals are controlled to be applied on different electrode layers during the transfer process, and different electrical signals applied on different electrode layers can produce different transfer efficiencies for proteins of different sizes. The overall structure of the solution disclosed in US8721860B2 is complicated, and the control of the transfer process is relatively complicated.

Summary of the invention

One of the objects of the present invention is to effectively avoid the temperature of the transfer unit during transfer Too high

Still another object of the present invention is to improve the transfer quality.

Still another object of the present invention is to ensure transfer efficiency without substantially increasing the transfer time.

In order to achieve at least one aspect or other objects of the above objects, the present invention provides the following technical solutions.

According to a first aspect of the invention, there is provided a biotransfer device comprising a control unit and one or more transfer units, the control unit being configured to: a first plate electrode layer to one or more of the transfer units Transmitting, with the second plate electrode layer, a unipolar electrical signal that periodically fluctuates according to a predetermined amplitude, wherein a ratio of an instantaneous power corresponding to a highest point of the predetermined amplitude to an instantaneous power corresponding to a lowest point of the predetermined amplitude is greater than or Equal to 2.

The biotransfer device of the invention is advantageous for maximizing the transfer work efficiency, ensuring the transfer efficiency, reducing the temperature of the transfer unit during the transfer process, reducing heat generation, and improving transfer quality.

A biotransfer device according to an embodiment of the present invention, wherein the undulating unipolar electrical signal comprises a square wave voltage signal with adjustable duty ratio, or an intermittent sine wave signal, a triangular wave signal, a sawtooth wave signal And at least one of the step signals. The unipolar electrical signals of this embodiment are easy to generate and the associated parameters are easy to control.

A biotransfer device according to an embodiment of the present invention, wherein the frequency of the unipolar electrical signal is greater than or equal to 1 Hz and less than or equal to 100 Hz, or greater than or equal to 5 Hz and less than or equal to 20 Hz. In the frequency range of this embodiment, it is more advantageous to avoid the continuous heat generation and temperature rise of the transfer unit, further improving the transfer quality.

A biotransfer device according to an embodiment of the present invention, wherein a duty ratio of the unipolar electrical signal is from 1% to 99%, or from 30% to 60%. The unipolar electrical signal of this embodiment provides an adjustable duty cycle range.

A biotransfer device according to an embodiment of the present invention, wherein the biotransfer device further includes temperature information for measuring the first plate electrode layer and/or the second electrode layer in real time during transfer Temperature sensor

Wherein the temperature information is fed back to the control unit, and the control unit is further configured to dynamically adjust the duty cycle based on at least the temperature information during transfer. The biotransfer device of this embodiment can be dynamically based on temperature information feedback The adjustment of the duty ratio makes the adjustment of the unipolar electrical signal more accurate, and is more advantageous for maximizing the efficiency of the transfer work, further reducing the temperature of the gel layer and the carrier film, reducing the heat generation, and improving the transfer quality.

A biotransfer device according to an embodiment of the present invention, wherein the control unit is further configured to: adjust at least the duty ratio and/or the maximum instantaneous power to cause the transfer unit to be in a transfer process The temperature is below 60 ° C - 70 ° C. Therefore, it is possible to completely avoid coking of the gel layer or the like of the transfer process transfer unit.

A biotransfer device according to an embodiment of the present invention, wherein the transfer unit comprises:

The first plate electrode layer and the second plate electrode layer disposed substantially in parallel;

a first buffer medium layer between the first plate electrode layer and the second plate electrode layer, a gel layer containing a biomacromolecule, a carrier film, and a second buffer medium layer;

Wherein the biomacromolecule in the gel layer is electrophoresed to the carrier film by the applied unipolar electrical signal.

A biotransfer device according to an embodiment of the present invention, wherein a ratio of a resistance of the first buffer dielectric layer, the second buffer dielectric layer, and the carrier film to a resistance of the gel layer is less than or equal to 3. The arrangement of the transfer unit of the embodiment of the present invention is advantageous for further improving the transfer work efficiency.

A biotransfer device according to an embodiment of the present invention, wherein energy and/or information transmission is achieved between the control unit and at least one of the transfer units by non-contact electromagnetic coupling. Therefore, the structure of the transfer device of the embodiment of the invention can be designed to be more compact, easier to achieve overall waterproof, biological or chemical pollution, more convenient and flexible to use, and is very suitable for use in a biological laboratory.

A biotransfer device according to an embodiment of the invention, wherein the control unit comprises:

a control parameter generation module for generating an adjusted control parameter based on the measurement information or/and the transfer quality information;

a unipolar electrical signal generating module, configured to generate the unipolar electrical signal according to the adjusted control parameter;

The adjusted control parameter includes at least one of a waveform, a frequency, a voltage value, a current value, and a duty ratio of the unipolar electrical signal;

Wherein the measurement information is temperature information of the first plate electrode layer and/or the second plate electrode layer measured in real time during the transfer process, or is recorded during the transfer process And including at least one of a waveform, a frequency, a voltage value, a current value, and a duty ratio of the unipolar electrical signal, and temperature information of the first plate electrode layer and/or the second plate electrode layer Timing information. The biotransfer device of the embodiment of the present invention is advantageous in further reducing the work for heat radiation and temperature increase during the transfer process, thereby contributing to lowering the temperature of the transfer unit and improving the transfer quality.

According to a second aspect of the present invention, a biotransfer system is provided, comprising:

a plurality of the above-described biotransfer devices;

a cloud server coupled to a plurality of control units of the biotransfer device;

The cloud server is configured to include:

a history database for storing measurement information or/and transfer quality information records obtained by each of the plurality of biotransfer devices at each transfer process as historical data information;

a cloud computing module for calculating a control parameter for generating a current transfer process corresponding to one of the plurality of biotransfer devices based on the historical data information;

a sending module, configured to send the control parameter to a corresponding control unit of the biotransfer device;

Wherein the control parameter includes at least one of a waveform, a frequency, a voltage value, a current value, and a duty ratio of the unipolar electrical signal; the measurement information is that the first tablet is measured in real time during a transfer process Temperature information of the electrode layer and/or the second plate electrode layer, or a waveform, a frequency, a voltage value, a current value, and a duty ratio including the unipolar electrical signal recorded during the transfer, and Timing information of at least one of temperature information of the first plate electrode layer and/or the second plate electrode layer.

The biotransfer system of the invention can quickly determine the control parameters conforming to the current transfer process based on historical data information, and is easy to implement the self-learning function, greatly improving the user experience, and greatly reducing the professional skills and experience requirements of the user or the experimenter.

According to a third aspect of the invention, there is provided a control method for a biotransfer device comprising one or more transfer units, the transfer unit comprising a first plate electrode layer and the second a plate electrode layer, and a gel layer and a carrier film between the first plate electrode layer and the second plate electrode layer; wherein the first plate electrode layer and the first to the transfer unit The two plate electrode layers uniformly apply a unipolar electrical signal that periodically fluctuates according to a predetermined amplitude, wherein a ratio of an instantaneous power corresponding to a highest point of the predetermined amplitude to an instantaneous power corresponding to a lowest point of the predetermined amplitude is greater than or Equal to 2.

The control method of the biotransfer device of the present invention is advantageous in maximizing the transfer work efficiency, ensuring the transfer efficiency, reducing the temperature of the transfer unit in the transfer process, reducing heat generation, and improving transfer quality.

A control method according to an embodiment of the present invention, wherein the undulating unipolar electrical signal comprises a square wave voltage signal with adjustable duty ratio, or an intermittent sine wave signal, a triangular wave signal, a sawtooth wave signal, and a ladder At least one of the signals. The unipolar electrical signals of this embodiment are easy to generate and the associated parameters are easy to control.

According to an embodiment of the present invention, the fluctuation frequency of the unipolar electrical signal is greater than or equal to 1 Hz and less than or equal to 100 Hz, or greater than or equal to 5 Hz and less than or equal to 20 Hz. In the frequency range of this embodiment, it is more advantageous to avoid the continuous heat generation and temperature rise of the transfer unit, further improving the transfer quality.

According to an embodiment of the present invention, the control method further includes:

And generating an adjusted control parameter based on the measurement information or/and the transfer quality information, the adjusted control parameter including at least one of a waveform, a frequency, a voltage value, a current value, and a duty ratio of the unipolar electrical signal ;as well as

Updating the unipolar electrical signal according to the adjusted control parameter, and applying the updated unipolar electrical signal to the transfer unit. The control method of the biotransfer device of this embodiment can dynamically adjust the duty ratio based on the temperature information feedback, and the adjustment of the unipolar electrical signal is more accurate, which is more advantageous for maximizing the transfer work efficiency and further reducing the condensation. The temperature of the glue layer and the carrier film and the like reduce heat generation and improve transfer quality.

The above features and operations of the present invention will become more apparent from the following description and drawings.

DRAWINGS

The above and other objects and advantages of the present invention will be more fully understood from the aspects of the appended claims.

BRIEF DESCRIPTION OF THE DRAWINGS Figure 1 is a schematic view showing the structure of a biotransfer device in accordance with a first embodiment of the present invention.

Fig. 2 is a schematic view showing the structure of a transfer unit in the biotransfer device of the first embodiment shown in Fig. 1.

3 is a schematic diagram of a unipolar electrical signal according to an embodiment of the invention, wherein FIG. 3(a) is a square wave voltage signal with adjustable duty ratio, and FIG. 3(b) is a transformation of FIG. 3(a). Example.

4 is a schematic diagram of a unipolar electrical signal according to still another embodiment of the present invention, wherein FIGS. 4(a) and 4(b) are sinusoidal voltage signals, and FIGS. 4(c) and 4(d) are The triangular voltage signal, Fig. 4(e) and Fig. 4(f) are sawtooth voltage signals, and Fig. 4(g) and Fig. 4(h) are step voltage signals.

Figure 5 is a block diagram showing the functional blocks of a control unit of a biotransfer device in accordance with an embodiment of the present invention.

Figure 6 is a graph showing the relationship between the resistance ratio and the transfer effective work.

Fig. 7 is a graph showing the calculated transfer effective work distribution.

Fig. 8 is a schematic view showing the control principle of the biotransfer device of the embodiment of the present invention.

Figure 9 is a schematic view showing the structure of a biotransfer device in accordance with a second embodiment of the present invention.

Figure 10 is a schematic view showing the structure of a biotransfer device in accordance with a third embodiment of the present invention.

11 is a schematic structural view of a non-contact electromagnetically coupled power supply module used in a biotransfer device according to an embodiment of the present invention.

Fig. 12 is a schematic structural view of a biotransfer system of a first embodiment formed based on the biotransfer device of the embodiment shown in Fig. 9.

Figure 13 is a schematic structural view of a biotransfer system of a second embodiment formed based on the biotransfer device of the embodiment shown in Figure 10.

14 is a block diagram showing the structure of a cloud server of a biotransfer system according to an embodiment of the present invention.

detailed description

The invention will now be described more fully hereinafter with reference to the accompanying drawings in which FIG. However, the invention may be embodied in many different forms and should not be construed as being limited to the embodiments set forth herein. Rather, these embodiments are provided so that this disclosure will be thorough and complete. In the drawings, the same reference numerals are used to refer to the same elements or components, and the description thereof will be omitted.

In the drawings, the size of the transfer unit layers and regions are exaggerated for clarity, and the dimensions of the various layers in the transfer unit in the drawings do not limit the actual size and size ratio relationship of the respective layers.

Some of the block diagrams shown in the figures are functional entities and do not necessarily have to correspond to physically or logically separate entities. In some cases, software can be used to implement these functions. These functional entities can be implemented in one or more hardware modules or integrated circuits, or in some cases in different networks and/or processor devices and/or microcontroller devices, in still other cases. Functional entity.

1 is a schematic view showing the structure of a biotransfer device according to an embodiment of the present invention. The biotransfer device 10 of the embodiment of the present invention mainly includes a transfer unit 130 and a control unit 11, wherein the transfer unit 130 is a specific execution component of the transfer process, and the main function of the control unit 11 is to control the specificity of the transfer unit 130. The working process is specifically achieved by controlling the electrical signal applied to the transfer unit 130.

Fig. 2 is a view showing the structure of a transfer unit in the biotransfer device of the embodiment shown in Fig. 1. Referring to FIG. 2, the transfer unit 130 is mainly formed of a stacked structure 131 stacked between upper and lower electrodes and upper and lower electrodes, which specifically includes an upper plate electrode layer 1301 and a lower plate electrode layer 1302, and is located on the upper plate electrode layer 1301 and below. The upper buffer dielectric layer 131e between the plate electrode layers 1302, the gel layer 131d containing the biomacromolecules, the carrier film 131c, and the lower buffer dielectric layer 131b. The function of the transfer unit 130 is to apply an electric signal to the upper plate electrode layer 1301 and the lower plate electrode layer 1302, thereby generating an electric field E similar to the direction indicated by the dotted arrow in FIG. 2, the biomacromolecules in the gel layer 131d. Under the action of the electric field E, the biomacromolecule is transferred to the carrier film 131c in accordance with the electrophoresis movement in the direction indicated by the time limit arrow in Fig. 2, and the transfer process is completed.

In the following embodiments of the present invention, the transfer process of the protein will be exemplified, but it should be understood that the biomacromolecules in the gel layer 131d to be transferred are not limited to proteins, and may be other similar Biological macromolecules, such as DNA. When the biotransfer device 10 is used for transferring proteins, it is specifically a Western blotting device; when the biotransfer device 10 is used for transferring DNA, it is specifically a Southern blotting device.

It should be noted that the upper plate electrode layer 1301 and the lower plate electrode layer 1302 may be formed of various conductive materials, and the specific material type thereof is not limited. In the present invention, the "plate electrode layer" means a planar single layered electrode structure (for each laminated structure 131 of the transfer unit), and an electric signal is applied to the upper plate electrode layer 1301 and the lower plate electrode. When the layer 1302 is on, the same electrical signal is uniformly applied between the upper plate electrode layer 1301 and the lower plate electrode layer 1302, and a uniform electric field E as shown in FIG. 2 is formed between the upper plate electrode layer 1301 and the lower plate electrode layer 1302. The electric field E is uniformly applied to proteins of different sizes in the gel layer 131d, that is, proteins of different sizes act on the uniform electric field E for electrophoresis. Upper plate electrode layer of embodiment of the invention The lower plate electrode layer 1302 has a simple structure and is simple to prepare.

It should be noted that the "uniformly" application of the electric field means that the electric field of the electrophoretic movement of the biomolecule of the gel layer 131d is uniform and simultaneously acts with respect to all the biomolecules such as the protein of the gel layer 131d.

It should be noted that the principles of applying the electrical signal to the protein on the upper plate electrode layer 1301 and the lower plate electrode layer 1302 disclosed in the embodiments of the present invention are the same as the patent number US8721860B2, entitled "Protein Multi-blotting Method and Device". The principle of electric field application disclosed in the patent is completely different. In US8721860B2, different electrical layers are controlled to apply different electrical signals. Therefore, the same electrical signal cannot be uniformly applied to different layer electrodes of the composite electrode layer, and is differently sized for different sizes of proteins. Different electrical signals applied to different electrode layers that affect the electrophoretic motion can produce different transfer efficiencies for different sized proteins, respectively. Therefore, the electrical signal in US8721860B2 is not spatially uniformly applied to the upper and lower electrode layers.

Continuing with FIG. 2, in an embodiment, the upper plate electrode layer 1301 and the lower plate electrode layer 1302 are disposed in a relatively parallel manner, such that the upper plate electrode layer 1301 and the lower plate electrode layer 1302 are not only spatially uniform. A certain electrical signal is applied, and the distribution of the electric field E generated by the electrical signal in the left-right direction as shown in FIG. 2 is also substantially uniform.

Continuing with FIG. 2, in an embodiment, the upper buffer dielectric layer 131e is located between the gel layer 131d and the upper plate electrode layer 1301, and the lower buffer dielectric layer 131b is located between the carrier film 131c and the lower plate electrode layer 1302. They may be formed with a buffer gel or filter paper material that first has a conductive effect, also has a protective effect on the gel layer 131d and the carrier film 131c, and provides a buffer ion for transfer. The resistance buffer layer 131e and the lower dielectric layer 131b of the cushioning medium can be measured or estimated from the characteristics of the material selected for use, are recorded as R _e and R _b. Incidentally, the cushioning medium layer resistance R _e 131e and / or the lower dielectric buffer layer R _b 131b is depending on the material, temperature change and changes in the number of buffer varies.

Continuing with Figure 2, in one embodiment, the gel layer 131d can be a biomass membrane that has, for example, a gel having a gradient concentration of 4%-20% or 4%-12%, or at 4%-12 A uniform concentration of gel in the range of %, different sizes of proteins to be transferred may be electrophoresed in the gel layer 131d; the thickness of the gel layer 131d may range from 0.5 mm to 2.0 mm, for example, may be 1 mm; The resistance of layer 131d is measured or estimated from the properties of the material selected for use, the resistance of which is recorded as _Rd . The carrier film 131c may be, for example, a nitrocellulose film or a PVDF (polyvinylidene fluoride) film, the specific material type of which is not limited, and the electrical resistance of the carrier film 131c may be measured or estimated according to the characteristics of the material selected for use. The resistance is recorded as R _c . It should be noted that the resistance R _{d of the} gel layer 131d and/or the R _c of the carrier film 131c vary depending on the material, the amount of the buffer, and the change in temperature.

Applicants need to point out that in each layer of the laminated structure 131 of the transfer unit 130, particularly in the gel layer 131d, it is easy to cause denaturation at a high temperature, and therefore, it is necessary to control the transfer unit 130 during transfer (especially The temperature of the gel layer 131d) is, for example, controlled to be lower than 60 ° C to 70 ° C to ensure transfer quality. However, in the prior art, a constant voltage, a step-up voltage, or a constant current signal is applied to the transfer unit 130 for the transfer operation; Applicants have found that the temperature easily exceeds 60 ° C as the transfer process proceeds or ends. -70 ° C, for example, in the case of a 25 V constant voltage signal for 10 min, the glue in the transfer unit 130 may even be scorched; and, in order to increase the transfer rate of large proteins, a higher voltage or a larger current is applied. It is not always possible to improve the transfer efficiency, and it is easy to cause the temperature of the transfer unit 130 to be too high, and the transfer quality is difficult to be secured.

Continuing, as shown in FIG. 1, the control unit 11 in the transfer device 10 is configured to apply undulating unipolar electrical signals to the upper plate electrode layer 1301 and the lower plate electrode layer 1302 of the one or more transfer units 130, the present invention. The unipolar electrical signal is periodically fluctuating according to a predetermined amplitude, wherein a ratio of the instantaneous power corresponding to the highest point of the predetermined amplitude to the instantaneous power corresponding to the lowest point of the predetermined amplitude is greater than or equal to two. It should be noted that the specific size of the predetermined amplitude may be constant or variable, that is, the ratio of the instantaneous power of the highest point of the predetermined amplitude to the instantaneous power of the lowest point corresponding to the predetermined amplitude. In the case of greater than or equal to 2, the ratio may be constant or varied during a periodic fluctuation of the signal period, for example, relatively constant during one control period, between a plurality of control periods, The signal period of each control cycle is adjustable. The time interval for completing the above-defined fluctuation process is defined as one signal period of the unipolar electrical signal. For example, the time interval at which the highest or lowest point of the above predetermined amplitude occurs continuously is defined as one signal period of the unipolar electrical signal.

For example, if the unipolar electrical signal is a 10 Hz square wave signal, the peak voltage is 25 V and the valley voltage is 5 V at the beginning of the transfer, then the signal period of the unipolar electrical signal is 100 milliseconds, one signal period. The instantaneous power ratio of the inner extreme point is 25×25 /(5×5)=25; After the transfer operation continues for example, for 1 second, the control unit adjusts the control parameters, and the unipolar electrical signal is still a square wave signal, but the signal frequency of the unipolar electrical signal becomes 20Hz, the peak voltage becomes 20V, and the valley voltage becomes 5V. Then, the signal period of the unipolar electrical signal corresponds to 50 milliseconds, and the instantaneous power ratio of the extreme point in one signal period is 20×20/(5× 5) = 16; after the transfer operation is continued for, for example, 1 second, the control unit further adjusts the control parameters to further obtain different square wave signals, which are continuously followed by predetermined time intervals during the transfer operation (for example, the above) The one-second electrical circuit is controlled or adjusted, and the predetermined time interval (for example, one second) is the control cycle referred to in this application. The principle of adjusting the unipolar electrical signal during the transfer operation based on the control period will be specifically described later.

In one embodiment, the instantaneous power corresponding to the highest point of the predetermined amplitude may be referred to as the maximum instantaneous power within the signal period, and the instantaneous power corresponding to the lowest point of the predetermined amplitude may be referred to as the minimum instantaneous period within the signal period. Power; the ratio of the maximum instantaneous power to the minimum instantaneous power during each signal period of the unipolar electrical signal is greater than or equal to about two, and in one embodiment, the ratio is greater than or equal to, for example, 2.5. It should be understood that, in each signal period, the unipolar electrical signal defines the amplitude of the undulation change in such a manner that the ratio of the maximum instantaneous power to the minimum instantaneous power in each signal period is greater than or equal to 2, in the corresponding waveform. The high point or peak (i.e., the highest point of the predetermined amplitude) corresponds to the maximum instantaneous power, and the low point or trough of the corresponding waveform (the lowest point of the predetermined amplitude) corresponds to the minimum instantaneous power.

Specifically, the unipolar electrical signal may be a voltage signal or a current signal, and the unipolar electrical signal may be specifically, but not limited to, a square wave signal, a sine wave signal, a sawtooth wave signal, or a staircase signal, or For their combination; the unipolar electrical signal can be continuous or intermittent. The waveform, frequency, voltage value, current value, and duty ratio of the unipolar electrical signal are important features or parameters that reflect the unipolar electrical signal. For example, the unipolar electrical signal is specifically a square wave voltage signal with adjustable duty ratio, the square wave voltage signal is relatively easy to control and easy to generate, and the duty ratio of the square wave voltage signal can be adjusted by adjusting the pulse width.

3 is a schematic diagram of a unipolar electrical signal according to an embodiment of the invention, wherein FIG. 3(a) is a square wave voltage signal 91 with adjustable duty ratio, and FIG. 3(b) is FIG. 3(a). An example of a transformation. As shown in FIG 3 (a), the peak voltage (i.e., high) is V _p, the low level of 0V, V _p is greater than or square wave voltage signal is equal to 91 and less than or equal to 1V 30V (e.g. 20V), the corresponding V _p the peak current on the transfer unit 130 is greater than or equal to the generated 0.1A and less than or equal to 10A (eg. 6A); corresponding to the signal V _p of each cycle can generate a maximum instantaneous power, low power corresponding to each signal period At the level, the minimum instantaneous power can be generated. The signal period T of the square wave voltage signal 91 can be adjusted, even in the same transfer process, specifically by controlling the frequency of the square wave voltage signal 91 to control its signal period T; in one embodiment, The frequency of the wave voltage signal 91 is greater than or equal to 1 Hz and less than or equal to 100 Hz, or for example greater than or equal to 5 Hz and less than or equal to 20 Hz, such as 10 Hz. The duty cycle of the square wave voltage signal 91 is also adjustable, with a duty cycle ranging from 1% to 99%, or for example from 30% to 60%.

As shown in FIG. 3(b), the square wave voltage signal 91' is a modified embodiment of the square wave voltage signal 91. In the square wave voltage signal 91', the voltage peak, the low level, the signal period T and/or Air ratios, etc. can vary. By setting the level of the high level Vp and the low level, the ratio of the maximum instantaneous power to the minimum instantaneous power in each signal period can be controlled to be greater than or equal to two. It is to be understood that the ratio of the maximum instantaneous power to the minimum instantaneous power in each signal period is not necessarily substantially constant.

4 is a schematic diagram of a unipolar electrical signal according to still another embodiment of the present invention, wherein FIGS. 4(a) and 4(b) are sinusoidal voltage signals, and FIG. 4(c) and FIG. 4(d) As a triangular voltage signal, FIG. 4(e) and FIG. 4(f) are sawtooth voltage signals, and FIG. 4(g) and FIG. 4(h) are step voltage signals.

FIG. 4 (a), the signal 92 is a sine half wave voltage signal, which is an intermittent signal, corresponding to a peak voltage V _p at the maximum instantaneous power can be generated in each cycle of the signal, corresponding to zero level may be generated at the Minimum instantaneous power. FIG. 4 (b), the signal 92 'is a full-wave sine wave voltage signal, but a minimum voltage greater than or equal to 0, which is a continuous electrical signal, corresponding to a peak voltage V _p of the signal may be generated in each cycle The maximum instantaneous power, corresponding to the minimum voltage, produces the minimum instantaneous power per signal period.

FIG. 4 (c), the signal is a triangular wave voltage signal 93, which is an intermittent signal, corresponding to a peak voltage V _p at the maximum instantaneous power can be generated in each cycle of the signal, corresponding to the zero level at the minimum instantaneous power may be generated . FIG. 4 (d), the signal 93 'is also a triangular wave voltage signal, but it is a continuous electrical signal, corresponding to a peak voltage V _p at the maximum instantaneous power can be generated in each cycle of the signal, corresponding to the minimum voltage (and which Not limited to 0), the minimum instantaneous power in each signal period can be generated.

FIG. 4 (e) and (f), the signal 94 and 94 'are both sawtooth wave voltage signal, which is an intermittent signal, corresponding to a peak voltage V _p at the maximum instantaneous power can be generated in each cycle of the signal, corresponding to The minimum instantaneous power can be generated at the 0 level.

As shown in FIG. 4(g), the signal 95 is a step voltage signal, which is an intermittent signal, wherein the voltage waveform changes in a stepwise manner when the voltage waveform changes, and the number of steps changes in each signal period, each time. magnitude of the voltage step change were not restrictive; the corresponding peak voltage V _p at the maximum instantaneous power can be generated in each cycle of the signal, corresponding to the zero level at the minimum instantaneous power can be generated. FIG. 4 (h), the signal 95 'is also a step voltage signal, a low level but greater than 0 volts, the voltage corresponding to the peak value V _p at the maximum instantaneous power can be generated in each cycle of the signal, may be generated at a voltage corresponding to the minimum Minimum instantaneous power per signal period.

As shown in FIG. 4 above embodiment unipolar electrical signal, which is a periodic characteristic; for a voltage signal, and the signal voltage within each cycle, the voltage V _p and the minimum peak in each signal period is provided The magnitude of the voltage allows the maximum instantaneous power to minimum instantaneous power ratio in each signal period to be greater than or equal to 2; for current signals, each signal period can be achieved by setting the current peak and minimum current values for each signal period. The ratio of the maximum instantaneous power to the minimum instantaneous power within is greater than or equal to two. It should be understood that the specific waveform of the undulating unipolar electrical signal is not limited to the above embodiment, and may be specifically selected according to specific application requirements. For unipolar electrical signals such as sinusoidal, triangular, sawtooth or stepped undulating variations, their respective duty cycles can be defined, for example, the maximum voltage in each signal period corresponds to the average of the minimum voltages. The ratio of the time period greater than the average value to the signal period T is defined as the duty ratio; thus, similarly to the square wave voltage signal 91, their duty ratios can also be adjusted, for example, the range of the duty ratio is 1%-99%, or for example 30%-60%.

Continuing with FIG. 1, the control unit 11 is configured to generate the voltage signal of the above embodiment (to generate a square wave voltage signal 91 as shown in FIG. 3(a), which specifically includes the waveform generator 110, the controller 150, and the power supply. Module 190, Human Machine Interface (HMI) 170, real time clock 160, voltage and/or current sensing component 120, and the like. Wherein, the controller 150 is a core component of the control unit 11, which is capable of outputting control parameters to the waveform generator 110 to generate corresponding unipolar electrical signals, such as square wave voltage signals; the controller 150 can have measurements, calculations, and controls Even the storage function, its specific working principle will be revealed in detail later.

The voltage and current detecting component 120 is configured to detect the voltage U and/or the current I applied to the transfer unit 130 in real time and feed it back to the controller 150 as measurement information.

The power module 190 is used to supply power to the biotransfer device 10. For example, it can provide AC power or DC power to the waveform generator 110 to generate a unipolar electrical signal of a corresponding waveform, and can also provide a low voltage DC power supply. The controller 150, and through the controller 150, can supply power to the real time clock 160, the voltage and current detecting component 120, and the like that are electrically connected thereto.

The real-time clock 160 can provide the current actual time to the waveform generator 110 and the controller 150. Based on the actual time information, the square wave signal having the corresponding frequency (or signal period), duty ratio, and the like can be controlled to be generated; The measurement information received by the device 150 or the self-measured information, such as the voltage or current information fed back by the voltage and current detecting component 120, combined with the actual time information provided by the real-time clock 160, generates measurement information having a corresponding time stamp, so that a pair can be obtained. The time series information of the information should be measured, and the time series information of the measurement information will be used in the calculation process of the control algorithm. Specifically, the real time clock 160 may be embedded in the controller 150.

Wherein, the human-computer interaction interface 170 is used to implement interaction with the user, for example, to implement a user selection or setting of a transfer option, a function of transferring parameters, and a function of starting or stopping the transfer process, and providing user feedback to the controller 150. Function, as well as the ability to present status information to the user during the transfer process. The human-machine interaction interface 170 is not limited to being integrally mounted on the bio-transfer device 10, and may be disposed, for example, separately from the main body of the bio-transfer device 10.

In an embodiment, the control unit 11 further includes a communication unit 180 coupled to the controller 150, by which the control unit 11 can be enabled with external smart terminals (eg, tablets, smart phones, etc.) and/or Alternatively, a cloud computing server or the like may be connected, and some functions of the control unit 11 may be implemented by an external device. For example, the human-machine interaction interface 170 may alternatively be implemented by an external tablet (IPAD) or the like.

Continuing with FIG. 1 , in an embodiment, in the biotransfer device 10 , the corresponding transfer unit 130 is further provided with a temperature sensor 140. During the transfer process, the temperature sensor 140 measures the temperature information of the transfer unit 130 in real time, for example, The temperature information of the upper plate electrode layer 1301 and/or the lower plate electrode layer 1302 is measured, and the measured temperature information substantially accurately reflects the temperature of the current gel layer 131d. The temperature information measured by the temperature sensor 140 can be fed back to the control unit 11 as measurement information, for example, to the controller 150. The temperature sensor 140 may be specifically integrated with the transfer unit 130.

FIG. 5 shows a control unit 11 of a biotransfer device according to an embodiment of the present invention. Schematic diagram of the functional module structure. As shown in FIG. 5 and FIG. 1, in an embodiment, the controller 150 controls the waveform generator 110 by outputting control parameters. Therefore, the control unit 11 is provided with a control parameter generation module 151, which can be disposed in the controller 150. Alternatively, it is implemented by the controller 150; the control unit 11 is also provided with a unipolar electrical signal generation module 111, which may be provided in the waveform generator 110 or implemented by the waveform generator 110. The control parameter generating module 151 is configured to generate an adjusted control parameter based on the measurement information or/and the transfer quality information, and the unipolar electrical signal generating module 111 is configured to generate a unipolar electrical signal according to the adjusted control parameter (for example, Square wave voltage signal, etc.).

Wherein the control parameter comprises at least one of a waveform, a frequency, a voltage value, a current value and a duty ratio of the unipolar electrical signal; the measurement information is that the upper plate electrode layer 1301 and/or are measured in real time during the transfer process Or temperature information of the lower plate electrode layer 1302, the measurement information being either a waveform including a unipolar electrical signal recorded during the transfer process (for example, a square wave, a sawtooth wave, a triangular wave or a sine wave, etc.), a voltage value Timing information of at least one of U, current value I and duty ratio, and temperature information of the upper plate electrode layer 1301 and/or the lower plate electrode layer 1302. In this way, it is possible to adjust the waveform, duty cycle, voltage value, current value and other parameters of the unipolar electrical signal in real time according to the feedback of the measurement information or/and the transfer quality information, thereby facilitating the maximization of the transfer work efficiency. While ensuring the transfer efficiency, the temperature of the gel layer and the carrier film is lowered and the heat generation is reduced, and the transfer quality is improved.

In an embodiment, the control unit 11 further comprises an input module 154 for receiving input transfer quality information, for example, obtaining a result for the transfer process at the end of the transfer process for a certain transfer unit 130 The quality is manually evaluated and input feedback, for example, input quality information is input from the human-machine interface 170 or an external device connected to the communication unit 180, and the input module 154 receives transfer quality information corresponding to each transfer process. The input module 154 can be disposed in the controller 150 or implemented by the controller 150.

When evaluating the transfer quality information of each transfer process, the user can evaluate according to a predetermined index or standard, and specifically can obtain the transfer quality information expressed by grade or by fraction, for example, based on the following Table 1 The print quality rating standard is used to determine the transfer quality information Qt.

Table I

Evaluation Criteria	Weights	Transfer quality score
Imprinting efficiency	50%	0-5
Gel layer condition	20%	0-2
Carrier film condition	20%	0-2
Imprinted form	5%	0-0.5
Buffer dielectric layer condition	5%	0-0.5
Transfer quality information (Qt)	100%	0-10

Among them, the elements for evaluating the transfer quality information include, but are not limited to, the transfer efficiency of the print, the condition of the gel layer, the condition of the carrier film, the form of the print, and the condition of the buffer medium layer, and the like, and the respective elements are given corresponding weights. For the transfer efficiency of the imprint, the scoring can be determined by reading the Marker information (for example, the number of imprints remaining on the gel layer 131d); for the condition of the gel layer, it can be burned according to the condition thereof, Curl, good, etc. to score; for the condition of the carrier film, it can be scored according to whether the condition corresponds to coking, curling, good, etc.; for the shape of the imprint, according to whether the condition corresponds to diffusion, clear Corresponding to the score; for the buffer medium layer condition, the score can be corresponding according to whether the condition corresponds to good coking or the like. Finally, based on the respective transfer quality scores, the overall transfer quality score, that is, the transfer quality information Qt, is calculated based on the respective weights.

It should be noted that the transfer quality information received each time corresponds to the corresponding transfer unit 130. Each of the plurality of substantially identical transfer units 130 can manually evaluate and input the corresponding transfer quality information Qt after each transfer operation.

It should be noted that the transfer quality information Qt is not limited to the determination of the transfer quality information according to the transfer quality scoring standard exemplified above, and the specificity may be adjusted according to the actual application, thereby helping the user to obtain the transfer quality as much as possible. information.

In an embodiment, the control unit 11 further includes a storage module 153 for recording measurement information or/and transfer quality information obtained for each transfer process. The storage module 153 may be disposed in the controller 150 or implemented by the controller 150. The measurement information may be various information obtained by real-time measurement of the transfer process, for example, the temperature of the transfer unit 130, the timing information described above, and the like. The measurement information or/and the transfer quality information recorded by the storage module 153 can be called by the control parameter generation module 151, which of course can also be selectively transmitted by the communication unit 180 to the external device to which it is connected.

In an embodiment, the storage module 153 may also store corresponding control parameters, such as generated by the control parameter generation module 151, corresponding to the transfer unit 130. After a certain transfer When the transfer unit 130 of the process is the same as the transfer unit 130 of the historical transfer process, the control parameter generation module 151 can directly call the control parameters stored in the historical transfer process stored by the storage module 153, thereby facilitating the rapid completion. The experimental design of the secondary transfer process (for example, including the setting of parameters), the generated undulating unipolar electrical signal will also be applied to the secondary transfer process, facilitating the rapid, high quality completion of the transfer process.

The biotransfer device 10 of the embodiment of the present invention is capable of applying a unipolar electrical signal that periodically changes to the transfer unit 130, and the maximum instantaneous power and the minimum instantaneous power in each signal period of the unipolar electrical signal. The ratio is greater than or equal to about 2; at the peak time period or time point corresponding to the maximum instantaneous power, the transfer unit 130 does not continue due to the valley time period or time point at which the power of the voltage or current drop is small in its subsequent time. When operating in a high power state, there is no possibility of a continuous superheat temperature rise like a constant current or a constant piezoelectric signal, and therefore, the magnitude of the voltage or current applied to the gel layer 131d of the transfer unit 130 can be substantially unrestricted. In this way, on the one hand, the large instantaneous power of the peak time period or time point can effectively promote the electrophoresis movement of biological macromolecules (especially large masses) such as proteins, ensuring and even improving the transfer efficiency; on the other hand, the trough time The smaller instantaneous power at the segment or time point can effectively prevent the temperature of the transfer unit 130 from rising due to ohmic heat, and the resistance of the transfer unit 130 is also Continues to increase due to heat generation, the temperature of the transfer unit 130 or reducing resistance, also the peak power period of time or point in time in a minor proportion work (i.e., W _Tm) for transferring heat radiation unit 130 and the temperature is increased, A large proportion of work (ie, transfer effective work _WEm ) is used to promote electrophoretic motion of biological macromolecules such as proteins. That is to say, during one signal period or multiple signal periods of the undulating unipolar electrical signal, the setting of the valley or time point of the smaller power is beneficial to increase the peak time period or time point of the larger power. The transfer performs work efficiency, thereby improving the transfer work efficiency of the unipolar electrical signal as a whole, and the transfer efficiency can be ensured or even improved, and the transfer unit 130 has a slow temperature increase and a good transfer quality during the transfer process.

The above technical effects of the biotransfer device 10 of the embodiment of the present invention will be described below by comparison test.

In the comparison test, the square wave voltage signal and the constant voltage signal generated by the biotransfer device 10 of the embodiment shown in FIG. 1 are subjected to a transfer effect comparison, wherein the square wave voltage signal is at a high level (ie, the peak voltage Vp). The same as the voltage of the constant voltage signal and the transfer operation time are the same, and the transfer unit 130 in which the square wave voltage signal and the constant voltage signal act is also the same. Need to say It is obvious that in the process of applying a constant voltage signal for the transfer operation, it is necessary to carry out a current limiting corresponding thereto, otherwise an excessive current is likely to be generated to burn the gel layer and the carrier film.

In the first comparison test, each of the two gel layers 131d juxtaposed in the transfer unit 130 was simultaneously subjected to a transfer operation, and the specific parameters for the two tests for comparison are shown in Table 2 below.

Table II

Wherein, the duty ratio of the applied square wave voltage signal is 40%, and when the high level is corresponding, the measured current peak value is 6A in the initial stage of the transfer process, and the measured current peak value is at the end of the transfer process. For 2A, the equivalent value is 4A; therefore, the average power during transfer is: 25V × 4A × 0.04s × 10/1s = 40W; the total power consumption for heat generation is: 40W × 60s × 15 = 36kJ. Correspondingly, the temperature of the gel layer (measured by temperature sensor 140) is about 65 °C. It should be noted that, in the above calculation process, when the low level is 5V, since the current is very small, the average power is also very small, and the generated power for heating is very small relative to the high level stage. Therefore, in the total use Ignore processing in the calculation of the power consumption of heat.

As an alignment, in the transfer process corresponding to the constant voltage signal, since the current limit is 2.5 A, the actual applied voltage measured at the initial stage of the transfer process is 18 V, which is measured at the end of the transfer process. The actual voltage applied is 24V; the equivalent value is 21V; therefore, the average power during transfer is: 21V × 2.5A = 52.5W; the total power consumption for heat generation is: 52.5W × 60s × 15 = 47kJ. Correspondingly, the temperature of the gel layer (measured by temperature sensor 140) is approximately 78 °C.

In the second comparison test, each test simultaneously performs a transfer operation on the 4 gel layers 131d juxtaposed in the transfer unit 130, and the specific parameters for the two tests for comparison are shown in Table 3 below.

Table 3

Wherein, the duty ratio of the applied square wave voltage signal is 40%, and when the high level is corresponding, the measured current peak value is 12A in the initial stage of the transfer process, and the measured current peak value is at the end of the transfer process. For 6A, the equivalent value is 4A; therefore, the average power during transfer is: 25V × 9A × 0.04s × 10/1s = 90W; the total power consumption for heat generation is: 90W × 60s × 15 = 81kJ. Correspondingly, the temperature of the gel layer (measured by temperature sensor 140) is about 72 °C. It should be noted that, in the above calculation process, when the low level is 5V, since the current is very small, the average power is also very small, and the generated power for heating is very small relative to the high level stage. Therefore, in the total use Ignore processing in the calculation of the power consumption of heat.

As an alignment, in the transfer process corresponding to the constant voltage signal, since the current limit is 5 A, the measured actual applied voltage is 15 V at the initial stage of the transfer process, and the measured application is applied at the end of the transfer process. The actual voltage is 25V; the equivalent value is 20V; therefore, the average power during the transfer process is: 20V × 5A = 100W; the total power consumption for heat generation is: 100W × 60s × 15 = 90kJ. Correspondingly, the temperature of the gel layer (measured by temperature sensor 140) is approximately 86 °C.

From the above first and second comparison tests, it can be found that less work is converted into heat during the transfer process of the corresponding square wave voltage signal, and the transfer operation can also be completed, and the gel layer temperature is also lower. The transfer quality is relatively better. This is because a lower proportion of energy is converted into heat during the transfer process, and a higher proportion of energy is used in the transfer process to achieve electrophoretic movement of the protein, that is, using a square wave voltage signal with respect to a constant voltage signal. High transfer work efficiency.

The following specific examples theoretically explain the reason why the transfer operation using the square wave voltage signal has a higher transfer work efficiency.

I. First, assume that the voltage applied to the laminated structure of the transfer unit 130 (including the upper buffer dielectric layer 131e, the gel layer 131d, the carrier film 131c, and the lower buffer dielectric layer 131b) is U, the upper buffer dielectric layer 131e, and the condensation The thickness of the adhesive layer 131d, the carrier film 131c and the lower buffer dielectric layer 131b are respectively d _e , d _d , d _c , d _b , and the resistivities are ρ, ρ _d , ρ _c and ρ _b , respectively, and the area (perpendicular to the electric field) The cross-sectional area of E is S, and the mobility of the target biomacromolecule in the gel layer 131d and the carrier film 131c is m _d and m _{c , respectively} . The above voltage U, thickness d and area S can be regarded as constant values during the transfer process, but their resistivities ρ _e , ρ _d , ρ _c and ρ _b vary during the transfer process. The change in resistivity is dependent on material properties and temperature, which is due to changes in temperature and/or material composition. The mobility rates m _d and m _c depend on the size and shape of the target biomacromolecule, and the structure of the gel layer 131d and the carrier film 131c (e.g., the size of the pores) and the material, and the upper buffer dielectric layer 131e and the lower buffer dielectric layer 131b. Chemical and physical characteristics, etc.

Ⅱ. 131d gel layer, the carrier film 131c, 131e on the buffer layer and the lower dielectric layer is buffered media of resistance R _d 131b, R _c, R _b and R _e may be by the following relationship (1-1-1), (1 -1-2), (1-1-3), (1-1-4) respectively indicate:

Wherein R _d , R _c , R _b and R _e are the electrical resistances of the gel layer 131d, the carrier film 131c, the upper buffer dielectric layer 131e and the lower buffer dielectric layer 131b, respectively.

III. Further, according to Ohm's law, the voltage U _d applied to the gel layer 131d and the electric field strength E _d can be expressed by the following relations (1-2-1) and (1-2-2), respectively:

Here, the voltage U _d is a voltage applied to the gel layer 131d, and E _d is an electric field intensity applied to the gel layer 131d.

IV. Further, the transfer rate v of the target biomacromolecule is further expressed based on the following relationship (1-3):

Where m is the mobility of the biomacromolecule.

. Ⅴ Suppose, m _d >> m _c, thereby, m _c t during the transfer time can be ignored, then the effective power transfer target biomacromolecule W _em can be calculated based on the following relation (1-4):

Among them, _Wem is the transfer effective work, the larger the transfer effective work ratio, the greater the transfer work efficiency; q is the charge of the target biomacromolecule, m is basically equal to m _d . Suppose q, t, m, and _d d is a constant value, taking into account the resistance _{R d, R c, R b} , R _e, and typical values generally in the range of 5-20 ohms, (1-4) from the above relation It is basically determined that the relationship between the transfer effective work _Wem (see the ordinate) and the resistance ratio (R _c + R _b + R _e ) / R _d (see the abscissa) is shown in Fig. 6. Therefore, the resistance ratio (R _c + R _b + R _e ) / Rd can be controlled to be less than or equal to 3 to increase the transfer effective work _Wem , thereby improving the transfer work efficiency.

VI. Taking the square wave voltage signal as an example, assuming that the duty ratio is μ, which can be set in the range of 1% to 99%, the relationship (1-5) can be further obtained based on the above relationship (1-4). :

. Ⅶ Since 131e on the buffer layer and the lower dielectric layer 131b buffered media resistance R _b and R _e depends significantly on the temperature T, the power consumption for which the generated heat; 131e on the buffer layer and the lower dielectric layer 131b, respectively buffered media The resistance at temperature T can be expressed by the following relations (1-6-1) and (1-6-2), respectively:

R _b (T)=R _b ·[k·(μ-η)2+1] (1-6-1)

R _e (T)=R _e ·[k·(μ-η) ² +1] (1-6-2)

Wherein R _b (T) and R _b (T) represent the resistance of the upper buffer dielectric layer 131e and the lower buffer dielectric layer 131b under the temperature T condition, respectively; k is a heating coefficient (for example, it can be estimated as 10); η is occupied a threshold value indicating that the duty ratio of the square wave voltage signal is equal to the duty cycle threshold, the temperature of the transfer process is no longer increased, for example, it may be equal to 20%; and R _b and R _e are upper buffer dielectric layers, respectively The resistance of 131e and lower buffer dielectric layer 131b before the start of transfer.

VIII. Based on the above relations (1-5), (1-6-1) and (1-6-2), and assuming R _c <<R _b , the change of _Rd in the transfer process is much smaller than R _b and The change of R _e can obtain the following relation (1-7) regarding the transfer effective work _Wem :

Wherein the value of k = 10, η = 20% , and the resistance _{R d, R c, R b} , R _e, and typical values typically 5-20 ohm, based on the above relational expression (1-7) may be simulated A transfer effective work profile as shown in Fig. 7 is obtained, wherein the higher the gradation, the smaller the transfer effective work _WEm , and vice versa.

As can be seen from Fig. 7, the square wave voltage signal having a duty ratio of about 40% has a significantly good transfer work efficiency with respect to the constant voltage signal (i.e., the signal of the duty ratio μ = 100%).

Moreover, it should be pointed out that, as can be seen from FIG. 7, in most cases, when the duty ratio μ is equal to about 50%, the transfer effective work _WE is a watershed, that is, the transfer effective work or the transfer work efficiency is relatively The peak value is reached; for the square wave signal used in the numerical simulation (such as the square wave signal shown in Figure 3(a)), the 50% duty cycle is equivalent to 50% of the full power output (the full power output corresponds to the duty cycle). 100% power output), that is, for other waveforms of unipolar electrical signals, it is necessary to achieve 50% or less (including 50%) full power output by adjusting the duty ratio in the range of 1%-99%. . If the ratio of the maximum instantaneous power P _max to the minimum instantaneous power P _{min in the} signal period is less than 2, it is assumed that the power output P is equivalently calculated based on the following relation (1-8):

P=μP _max +(1-μ)P _min (1-8)

Even if the duty ratio μ = 0%, the adjusted power output P cannot be output to 50% or less (including 50%) of the full power output. To this end, in an embodiment of the invention, the ratio of the maximum instantaneous power P _max to the minimum instantaneous power P _{mi n} in the signal period of the unipolar periodic signal is greater than or equal to about 2, that is, unipolar The electrical signal periodically fluctuates according to a predetermined amplitude, and the ratio of the instantaneous power of the highest point of the predetermined amplitude to the instantaneous power of the lowest point of the defined amplitude is greater than or equal to 2; thus, the power output P can be achieved by adjusting the duty ratio μ 50% or less of the full power output, that is, the transfer effective work or the transfer work efficiency can be relatively peaked.

It should be understood that if the unipolar electrical signal is voltage information, such as the square wave voltage signal as shown in FIG. 3(a), the ratio of the maximum instantaneous voltage in the set signal period to the minimum instantaneous voltage is greater than or equal to about

The ratio of the large instantaneous power P _max to the minimum instantaneous power P _{mi n can} be greater than or equal to about 2; if the unipolar electrical signal is current information, the ratio of the maximum instantaneous current _{x to the} minimum instantaneous current in the set signal period is greater than or equal to approximately

Greater than can be achieved instantaneous power P _max and the minimum instantaneous power P _{mi n} is greater than or equal to about 2.

Fig. 8 further exemplifies the control principle of the biotransfer device, and from this point of view, the technical effects of the biotransfer device 10 of the above embodiment of the present invention can also be exemplarily explained. The example illustrates how the duty ratio μ generated by the control parameter generation module 151 of the controller 150 and the same transfer are adjusted based on the measurement information of the temperature information T including the transfer unit 130 measured in real time during the primary transfer process. The unit 130 adjusts the control parameters based on the transfer quality information Qt during the multiple transfer process, thereby maximizing the transfer work efficiency. The square wave voltage signal will be described below by way of example with reference to FIG.

As shown in FIG. 8, in the first control loop process 191 of a certain transfer operation process, it corresponds to the mth transfer process of a certain transfer unit 130 (each transfer process may include a plurality of first controls) In the loop process 191, that is, including a plurality of control periods, the transfer effective work _{WEm is} maximized by dynamically adjusting the duty ratio μ of each control period in real time.

First, the transfer effective work _{Wem is} calculated by the following relation (2-1):

W _em =W _m -W _Tm (2-1);

Where W _m represents the total work done by the square wave voltage signal during transfer, and W _Tm represents the work consumed for heat radiation and temperature increase during the transfer process.

Further, W _m can be calculated by the following relation (2-2):

Where Δt is the time interval of the first control loop process 191, which corresponds to one control period, one control period is greater than or equal to one signal period, for example, it corresponds to a plurality of signal periods having unipolar electrical signals; m represents the mth In the secondary transfer process, U _m represents the voltage amplitude measured by the mth transfer process (if the minimum voltage of the unipolar electrical signal in the control period is equal to 0, the voltage amplitude represents the voltage peak), and I _i represents the ith control The current amplitude obtained by the period measurement (if the minimum current in the control period is equal to 0, the current amplitude represents the peak value of the electric current), and μ _i represents the duty ratio of the electrical signal in the ith control period (the unipolar electricity in the control period) The duty cycle of the signal is constant, that is, both μ _i ), 1 ≤ i ≤ (n-1).

At the same time, W _Tm can be calculated by the following relation (2-3):

Wherein ξ _m represents the equivalent thermal emissivity of the transfer unit 130 during the mth transfer; C represents the specific heat capacity of the transfer unit 130; T _i and T _i-1 represent the i and i (i-1), respectively The temperature information of the obtained transfer unit 130 is measured by a control period.

Based on the above relations (2-1), (2-2), and (2-3), the following relation (2-4) can be obtained:

Therefore, maximizing the transfer effective work _WEm , that is, finding the maximum value of the _Wem relative variable duty ratio μ, can be expressed as the following relation (2-5):

The temperature information T and the current information I measured in the first to (n-1)th control periods based on the square wave voltage signal are input to the above relation (2-5), and are calculated in the first to W _em maximized so that the duty ratio corresponding to the first (n-1) th control period, and the duty ratio of the duty output as [mu] _n the n-th control cycle, i.e., corresponding to the n-th control period The duty ratio μ _n in the control parameter is used, and the waveform generator 110 adjusts the duty ratio of the next control period (i.e., the nth control period) based on the duty ratio μ _n . It is foreseen that when the square wave voltage signal of the nth control period based on the duty ratio μ _n continues the transfer operation, the _Wem can continue to be maximized.

In the second control loop process 192, which is controlled based on the following relation (2-6):

Where i represents the i-th transfer process of the first to (m-1)th transfer processes, m is an integer greater than or equal to 2, and Q _ti represents transfer quality information corresponding to the i-th transfer process, W _ei represents the transfer effective work of the i-th transfer process, which can be calculated by the above relation (2-5). Based on the previous (m-1) and m-th input transfer quality information, the equivalent thermal emissivity ξ _m-1 can be obtained by fitting calculation. Further, the equivalent thermal emissivity ξ _{m-1 is} used as the intermediate input. The parameter, combined with the equivalent voltage U _m-1 of the voltage measured in the (m-1)th transfer process, is substituted into the relational formula (2-4), and the transfer effective work W _{em of} the mth transfer process is calculated. The transfer effective work _Wem is the maximum value when the relative equivalent heat radiation coefficient ξ is used as an independent variable.

The equivalent thermal radiation coefficient ξ obtained in the second control loop process 192 above may also be substituted into the relation (2-5) for obtaining the corresponding duty in the first control loop process 191. The specific parameters are obtained to obtain the maximum transfer effective work.

The above first control loop process 191 and second control loop process 192 are both for achieving the largest possible transfer work efficiency, thereby reducing the work consumed for heat radiation and temperature increase during the transfer process, which is advantageous for reducing The temperature of the transfer unit 130 increases the transfer quality.

Based on the above examples of the first control loop process 191 and the second control loop process 192, it will be understood that the control parameter generation module 151 of the controller 150 can automatically optimize other control parameters other than the duty cycle, such as frequency or period, voltage The value and/or current value or the like, the control parameter generation module 151 of the controller 150 has a self-learning function. Such a self-learning function can function in, for example, for example, a newly used biotransfer device 10, a new transfer unit 130 (eg, a new gel layer 131d, a carrier film 131c, and/or an upper and lower buffer dielectric layer). ), quickly setting its control parameters, from generating a suitable unipolar electrical signal; during a daily multiple transfer process of a certain biotransfer device 10 for the same transfer unit 130, based on the measurement information that has been obtained, transfer Quality information, etc. not only optimizes the control parameters of each transfer process, but also continuously improves the transfer quality. Moreover, this self-learning function can also greatly reduce the professional competence requirements of operators.

Specifically, the control parameter generation module 151 of the controller 150 may use a kernel fitting algorithm to perform calculation or optimize control parameters, and the kernel fitting algorithm uses an artificial neural network, which may specifically but not limited to a supervised learning network ( Supervised Learning Network), Hybrid Learning Network, Reinforcement Learning Network, Hopfield Network, Boltzmann Machine, Stochastic Neural Networks )Wait. The neural network data can be trained by setting different input variables to optimize different control parameters.

Figure 9 is a schematic view showing the structure of a biotransfer device in accordance with a second embodiment of the present invention. As shown in FIG. 9, the biotransfer device 20 is similarly provided with a waveform generator 110, a voltage and current detecting part 120, a transfer unit 130, a temperature sensor 140, and a control similar to those in the biotransfer device 10 shown in FIG. 1. One or more of the device 150, the real-time clock 160, the human-machine interaction interface 170, and the communication unit 180, and the detailed description thereof will not be repeated here, with respect to the biotransfer device 10, the power module provided in the bio-transfer device 20 190 has a relatively different implementation.

In an embodiment, as shown in FIG. 9, the power module 190 is coupled by electromagnetic coupling. Energy transfer is achieved, which includes a primary unit 191 and a secondary unit 192 such that the biotransfer device 20 can be made to contact power in a non-contact manner. Therefore, the structure of the biotransfer device 20 in the embodiment shown in FIG. 9 can be designed to be more compact, and it is easier to achieve overall waterproof, biological or chemical contamination (for example, reducing metal contamination of biological agents and the like due to conductive contact). ), more convenient and flexible to use, very suitable for use in biological laboratories.

Figure 10 is a schematic view showing the structure of a biotransfer device in accordance with a third embodiment of the present invention. As shown in FIG. 10, the biotransfer device 20 is similarly provided with a waveform generator 110, a voltage and current detecting part 120, a transfer unit 130, a temperature sensor 140, and a control similar to those in the biotransfer device 10 shown in FIG. 1. One or more of the device 150, the real-time clock 160, the human-machine interaction interface 170, and the communication unit 180, and details are not described herein again. The biotransfer device 20 mainly has the following differences with respect to the biotransfer device 10: (1) The biotransfer device 20 is structurally divided into a control end 310 and a transfer end 320, and the control end 310 and the transfer end 320 are separated. (2) The power module 190 has a relatively different implementation. In particular, energy and/or information transmission is achieved between the control terminal 310 and the transfer terminal 320 by non-contact electromagnetic coupling.

Specifically, as shown in FIG. 10, the control terminal 310 is mainly used to implement the function of the control unit, which is provided with a waveform generator 110, a voltage and current detecting component 120, and a controller similar to those in the biotransfer device 10 as shown in FIG. 1. 150. The real-time clock 160, the human-machine interaction interface 170, and the communication unit 180. The control terminal 310 is further provided with an alternating current-direct current (AC-DC) conversion unit 193 for supplying power to various components. The AC-DC conversion unit 193 is primarily used to convert the AC power of the grid to a corresponding level of DC power, which is part of the power module. The unipolar electrical signal (eg, a square wave voltage signal) generated by the control terminal 310 for application on the transfer unit 130 is wirelessly transmitted to the transfer end 320, for example, may be simultaneously transmitted to multiple (eg, x, x) Is an integer greater than or equal to 2) transfer ends 320 ₁ , 320 ₂ ... 320 _x . In addition to the secondary unit 192 provided with the transfer unit 130 and the power module, in each embodiment, a temperature sensor 140 is also integrally provided. Corresponding to each transfer end 320, a primary unit 191 is correspondingly disposed on the control terminal 310. For example, one control terminal 310 is provided with x (x≥2) primary units 191 ₁ , 191 ₂ ... 191 _x . Thus, on the one hand, the unipolar electrical signal generated by the control terminal 310 can be transmitted to the plurality of transfer ends 320 by non-contact electromagnetic coupling, that is, to achieve non-contact transfer of energy, and on the other hand, information generated by each transfer end 320. For example, the temperature information collected by the temperature sensor 140 is transmitted to the control terminal 310 through the non-contact electromagnetic coupling, that is, the information is transmitted non-contactly.

Figure 11 is a block diagram showing the structure of a non-contact electromagnetically coupled power supply module used in a biotransfer device in accordance with an embodiment of the present invention. The power module 190 in the biotransfer device 30 includes a primary unit 191 and a secondary unit 192, a primary coil 1911 and a magnetic axis 1912 are disposed in the primary unit 191, and a secondary coil 1921 is disposed in the secondary unit 192. The primary unit 191 is provided. And the secondary unit 192 can be separately wrapped, for example, by a plastic shell. When the magnetic axis 1912 of the primary unit 191 approaches or is inserted into the secondary coil 1921 of the secondary unit 192, non-contact electromagnetic coupling is implemented between them based on the principle of electromagnetic induction, so that the square wave voltage signal of the control terminal 310 can be The temperature information and the like of the transfer end 320 can also be transmitted to the control terminal 310 in the form of a digital square wave.

In the biotransfer device 30 of the embodiment shown in FIG. 10, the structure of the transfer end 320 can be designed to be more compact, and it is easier to achieve waterproof design, biological or chemical contamination (for example, reducing biological agents caused by conductive contact). It is more convenient and flexible to use, which is very suitable for application in biological laboratories.

Fig. 12 is a view showing the configuration of a biotransfer system of a first embodiment formed based on the biotransfer device of the embodiment shown in Fig. 9. As shown in FIG. 12, the biotransfer system includes a plurality of biotransfer devices 20, and their control units are coupled to the cloud server 90, so that the cloud server 90 and any one of the biotransfer devices 20 can be realized. Information or data transmission, for example, measurement information and/or transfer quality information obtained in the biotransfer device 20, etc., may be uploaded to the cloud server 90, and the cloud server 90 may also transmit information (eg, control parameters) to the corresponding biotransfer. Device 20.

In an embodiment, n (n≥2) biotransfer devices 20 ₁ , 20 ₂ . . . 20 _n may be simultaneously communicatively coupled to a portable smart terminal 175, for example, via communication unit 180 of biotransfer device 20 The portable smart terminal 175, the portable smart terminal 175 may specifically be an tablet (IPAD), a smart phone, etc., and the portable smart terminal 175 may implement the function of the human-machine interaction interface 170 of the bio-transfer device 20 in whole or in part, for example, by A corresponding APP application is installed thereon to effect interaction with each of the biotransfer devices 20, particularly the controller 150.

In an embodiment, the cloud server 90 can be communicatively coupled with m (m ≥ 2) portable smart terminals 175 ₁ ... 175 _m to implement any bio-transfer connected to the m portable smart terminals 175 ₁ ... 175 _m The printing device 20 is coupled; in yet another alternative embodiment, the cloud server 90 can also be in direct communication with the transfer device 20.

Fig. 13 is a view showing the configuration of a biotransfer system of a second embodiment formed based on the biotransfer device of the embodiment shown in Fig. 10. As shown in FIG. 13, the cloud server 90 of the biotransfer system is communicatively coupled with m (m≥2) portable intelligent terminals 175 ₁ ... 175 _m , each of which has n (n ≥ 2) biotransfers. The devices 30 ₁ , 30 ₂ ... 30 _{n are} communicatively coupled.

FIG. 14 is a block diagram showing the structure of a cloud server of a biotransfer system according to an embodiment of the present invention. As shown in FIG. 14, the cloud server 90 is provided with a history database 910, a cloud computing module 920, and a communication module 930; wherein the history database 910 can store data sent by each biotransfer device, and the history database 910 can have at least multiple The measurement information or/and the transfer quality information record obtained by each of the biotransfer devices 20 or 30 at each transfer process is stored as historical data information (for example, data stored by the storage module 153 of each biotransfer device). It can be sent to the historical database 910), so that as the transfer process continues, a large amount of historical data information can be formed, and more and more historical data information can provide more and more accurate and effective cloud computing. Large help; the cloud computing module 920 can perform cloud computing or big data analysis processing on the historical data, and specifically generate a current turn corresponding to one of the plurality of biotransfer devices (10 or 20 or 30) based on the historical data information. The control parameters of the printing process, thereby facilitating the use of historical data information to quickly determine the control parameters in accordance with the current transfer process, greatly improving the user experience .

It should be understood that the cloud computing module 920 can use various cloud technologies to enhance the robustness of the intelligent control during the calculation and analysis process, and the history database 910 can selectively store the measurement information corresponding to the transfer process with better transfer quality information. And / or control parameters, in the cloud computing process, which can accelerate the efficiency of self-learning to obtain the best control parameters and reduce the workload of cloud computing. In an embodiment, the cloud computing module 920 may also completely or partially replace the functions of the control parameter generation module 151 in the completion controller 150.

Continuing with FIG. 9, the control parameters obtained by the cloud computing module 920 can be sent to the control unit of the corresponding biotransfer device 10, 20 or 30 via the transmitting module 193. The biotransfer device 10, 20 or 30 can generate or update a corresponding unipolar electrical signal based on the control parameters.

It should be understood that, due to the introduction of the cloud server 90, the sharing of historical data information of many biotransfer devices is realized, and the calculation parameters of the historical data information are obtained by the cloud computing module 920, which is equivalent to realizing the setting control of each user. The experience of parameters is shared and exploited, greatly reducing the professional skills and experience of users or experimenters. begging.

It will be understood that when a component is "connected", "coupled" or "coupled" to another component, it can be directly connected, coupled, or coupled to the other component.

The above examples mainly illustrate the biotransfer device of the present invention, a control method therefor, and a biotransfer system formed based on a plurality of biotransfer devices. Although only a few of the embodiments of the present invention have been described, it will be understood by those skilled in the art that the invention may be practiced in many other forms without departing from the spirit and scope of the invention. Accordingly, the present invention is to be construed as illustrative and not restrictive, and the invention may cover various modifications without departing from the spirit and scope of the invention as defined by the appended claims With replacement.

Claims (24)

1. A biotransfer device comprising a control unit and one or more transfer units, wherein the control unit is configured to: a first plate electrode layer and a second plate electrode to one or more of the transfer units The layer applies a unipolar electrical signal that periodically varies in accordance with a predetermined amplitude, wherein a ratio of the instantaneous power corresponding to the highest point of the predetermined amplitude to the instantaneous power corresponding to the lowest point of the predetermined amplitude is greater than or equal to two.
2. The biotransfer device according to claim 1, wherein said predetermined amplitude is constant or varied during said periodic fluctuation of said monopolar electrical signal.
3. The biotransfer device according to claim 1, wherein the undulating unipolar electrical signal comprises at least one of a square wave signal, a sine wave signal, a triangular wave signal, a sawtooth wave signal, and a staircase signal. .
4. A biotransfer device according to claim 1, wherein said undulating unipolar electrical signal comprises a square wave voltage signal with adjustable duty cycle, or intermittent sinusoidal signal, triangular wave signal, sawtooth At least one of a wave signal and a staircase signal.
5. The biotransfer device according to claim 3 or 4, wherein the frequency of the unipolar electrical signal is greater than or equal to 1 Hz and less than or equal to 100 Hz, or greater than or equal to 5 Hz and less than or equal to 20 Hz.
6. The biotransfer device according to claim 3 or 4, wherein the duty ratio of the unipolar electrical signal is from 1% to 99%, or from 30% to 60%.
7. The biotransfer device according to claim 3 or 4, wherein the unipolar electrical signal is a voltage signal having a voltage peak greater than or equal to 1 V and less than or equal to 30 V, corresponding to the voltage peak at least one The peak current generated on the transfer unit is greater than or equal to 0.1 A and less than or equal to 10 A.
8. The biotransfer device according to claim 1, wherein said transfer unit comprises:

   The first plate electrode layer and the second plate electrode layer disposed substantially in parallel;

   a first buffer medium layer between the first plate electrode layer and the second plate electrode layer, a gel layer containing a biomacromolecule, a carrier film, and a second buffer medium layer;

   Wherein the biomacromolecule in the gel layer is applied to the unipolar electrical signal Electrophoresis to the carrier film.
9. The biotransfer device according to claim 8, wherein a ratio of a resistance of said first buffer dielectric layer, said second buffer dielectric layer, and said carrier film to a resistance of said gel layer is less than or equal to 3.
10. The biotransfer device according to claim 6, wherein said biotransfer device further comprises means for measuring said first plate electrode layer and/or said second electrode layer in real time during transfer Temperature sensor for temperature information;

    Wherein the temperature information is fed back to the control unit, and the control unit is further configured to dynamically adjust the duty cycle based on at least the temperature information during transfer.
11. The biotransfer device according to claim 10, wherein the control unit is further configured to: adjust the at least the duty ratio and/or the maximum instantaneous power to cause the transfer unit to be transferred The temperature of the process is below 60 ° C - 70 ° C.
12. The biotransfer device according to claim 1, wherein energy and/or information transmission is achieved between the control unit and at least one of the transfer units by non-contact electromagnetic coupling.
13. The biotransfer device according to claim 1, wherein the biotransfer device is a Western blotting device or a Southern blotting device.
14. The biotransfer device according to claim 1, wherein said control unit comprises:

    a control parameter generation module for generating an adjusted control parameter based on the measurement information or/and the transfer quality information;

    a unipolar electrical signal generating module, configured to generate the unipolar electrical signal according to the adjusted control parameter;

    The adjusted control parameter includes at least one of a waveform, a frequency, a voltage value, a current value, and a duty ratio of the unipolar electrical signal;

    Wherein the measurement information is temperature information of the first plate electrode layer and/or the second plate electrode layer measured in real time during the transfer process, or includes the monopole recorded during the transfer process Timing information of at least one of a waveform, a frequency, a voltage value, a current value, and a duty ratio of the electrical signal, and temperature information of the first plate electrode layer and/or the second plate electrode layer.
15. The biotransfer device according to claim 14, wherein said control The unit further includes:

    An input module for receiving the input transfer quality information;

    A storage module for recording the measurement information or/and transfer quality information for each transfer process.
16. A biotransfer system, comprising:

    a plurality of biotransfer devices according to any one of claims 1 to 15;

    a cloud server coupled to a plurality of control units of the biotransfer device;

    The cloud server is configured to include:

    a history database for storing measurement information or/and transfer quality information records obtained by each of the plurality of biotransfer devices at each transfer process as historical data information;

    a cloud computing module for calculating a control parameter for generating a current transfer process corresponding to one of the plurality of biotransfer devices based on the historical data information;

    a sending module, configured to send the control parameter to a corresponding control unit of the biotransfer device;

    Wherein the control parameter includes at least one of a waveform, a frequency, a voltage value, a current value, and a duty ratio of the unipolar electrical signal; the measurement information is that the first tablet is measured in real time during a transfer process Temperature information of the electrode layer and/or the second plate electrode layer, or a waveform, a frequency, a voltage value, a current value, and a duty ratio including the unipolar electrical signal recorded during the transfer, and Timing information of at least one of temperature information of the first plate electrode layer and/or the second plate electrode layer.
17. A control method for a biotransfer device including one or more transfer units, the transfer unit including a first plate electrode layer and the second plate electrode layer, and located at the a gel layer and a carrier film between a plate electrode layer and the second plate electrode layer; characterized by a first plate electrode layer and a second layer to one or more of the transfer units during transfer The plate electrode layer uniformly applies a unipolar electrical signal that periodically fluctuates according to a predetermined amplitude, wherein a ratio of an instantaneous power corresponding to a highest point of the predetermined amplitude to an instantaneous power corresponding to a lowest point of the predetermined amplitude is greater than or equal to 2.
18. The control method according to claim 17, wherein the undulating unipolar electrical signal comprises at least one of a square wave signal, a sine wave signal, a triangular wave signal, a sawtooth wave signal, and a staircase signal.
19. The control method according to claim 17, wherein said fluctuation changes The unipolar electrical signal includes a square wave voltage signal with an adjustable duty cycle, or at least one of an intermittent sine wave signal, a triangular wave signal, a sawtooth wave signal, and a staircase signal.
20. The control method according to claim 18 or 19, wherein the fluctuation frequency of the unipolar electrical signal is greater than or equal to 1 Hz and less than or equal to 100 Hz, or greater than or equal to 5 Hz and less than or equal to 20 Hz.
21. The control method according to claim 18 or 19, wherein the duty ratio of the unipolar electrical signal is from 1% to 99%, or from 30% to 60%.
22. The control method according to claim 18 or 19, wherein the unipolar electrical signal is a voltage signal having a voltage peak greater than or equal to 1 V and less than or equal to 30 V, corresponding to the voltage peak in at least one of The peak current generated on the transfer unit is greater than or equal to 0.1 A and less than or equal to 10 A.
23. The control method according to claim 21, wherein the control method further comprises:

    And generating an adjusted control parameter based on the measurement information or/and the transfer quality information, the adjusted control parameter including at least one of a waveform, a frequency, a voltage value, a current value, and a duty ratio of the unipolar electrical signal ;as well as

    Updating the unipolar electrical signal according to the adjusted control parameter, and applying the updated unipolar electrical signal to the transfer unit.
24. The control method according to claim 23, further comprising: before the generating step, further comprising:

    Measuring temperature information of the first plate electrode layer and/or the second plate electrode layer as the measurement information in real time during transfer; or

    Time-series information including at least one of a waveform, a frequency, a voltage value, a current value and a duty ratio, and temperature information of the unipolar electrical signal, wherein the temperature is recorded, during the transfer process The information is obtained by measuring the temperature of the first plate electrode layer and/or the second electrode layer in real time during the transfer process.

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