CN110249220B - Biological transfer apparatus, biological transfer system, and control method - Google Patents

Abstract

A biological transfer apparatus (10), a biological transfer system, and a control method. The biological transfer device (10) comprises a control unit (11) and one or more transfer units (130), wherein the control unit (11) is configured to: and applying a unipolar electric signal periodically fluctuating with a predetermined amplitude to the first and second plate electrode layers of one or more transfer units (130), wherein the 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 2. The biological transfer printing device has high transfer printing work efficiency, can ensure the transfer efficiency and has good transfer printing quality.

Description

Biological transfer apparatus, biological transfer system, and control method

Technical Field

The invention belongs to the technical field of biochemical analysis, relates to transfer printing of biological macromolecules, and particularly relates to a biological transfer printing device using a fluctuating unipolar electric signal, a biological transfer printing system and a control method of the biological transfer printing system.

Background

The bio-Transfer Device (Blot Transfer Device) utilizes the principle of electrophoresis to Transfer bio-macromolecules (e.g., proteins) in a gel layer containing bio-macromolecules onto a carrier film, and thus, the bio-Transfer Device generally includes a control unit for generating corresponding electrical signals applied to the gel layer to control the above-mentioned electrophoresis-based Transfer process, and the electrical signals are applied to electrodes of a Transfer unit including a laminated structure of the gel layer and the carrier film.

Taking the Western Blot Transfer (Western Blot Transfer) as an example, the molecular sizes of proteins during Transfer are usually not uniform, and the Transfer rates under the same electrical signal are not the same for different sized proteins, especially for relatively larger proteins. At present, it is generally considered that transfer efficiency and transfer quality in the bio-transfer device are proportional to a voltage applied to the transfer unit, and thus, the transfer efficiency is improved and good transfer quality is obtained by increasing the voltage applied to the transfer unit. However, this method, which simply increases the voltage applied to the transfer unit or extends the transfer time, has problems that the temperature of the transfer unit is easily too high and the transfer quality is poor, and the transfer efficiency cannot necessarily be ensured.

In the scheme for improving the transfer efficiency of the western blotting apparatus disclosed in U.S. Pat. No. 8721860B2, entitled "Protein Multi-blotting Method and Device", one of the upper and lower electrode layers at the transfer unit terminal of the Protein transfer apparatus is a multiple composite electrode layer, and different electrical signals are controlled to be applied to different electrode layers during the transfer process, so that different transfer efficiencies can be generated for proteins of different sizes by applying different electrical signals to different electrode layers. The solution disclosed in US8721860B2 is complex in overall structure and relatively complex in control of the transfer process.

Disclosure of Invention

One of the purposes of the invention is to effectively avoid the over-high temperature of the transfer unit in the transfer process;

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

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

To achieve at least one of the above objects or other objects, the present invention provides the following technical solutions.

According to a first aspect of the present invention, there is provided a biological transfer apparatus including a control unit and one or more transfer units, the control unit being configured to: and applying a unipolar electric signal which fluctuates periodically according to a predetermined amplitude to the first flat electrode layer and the second flat electrode layer of one or more of the transfer units, wherein the 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 2.

The biological transfer printing device is beneficial to realizing maximization of transfer printing acting efficiency, ensures the transfer efficiency, reduces the temperature of a transfer unit in the transfer printing process, reduces heat generation and improves the transfer printing quality.

According to an embodiment of the invention, the fluctuating unipolar electric signal includes a square wave voltage signal with an adjustable duty ratio, or at least one of an intermittent sine wave signal, a triangular wave signal, a sawtooth wave signal, and a staircase signal. The unipolar electric signal of this embodiment is easy to generate and the relevant parameters are easy to control.

According to one embodiment of the invention, the frequency of the unipolar electric signal is greater than or equal to 1Hz and less than or equal to 100Hz, or greater than or equal to 5Hz and less than or equal to 20Hz. In the frequency range of this embodiment, it is more favorable to avoid the transfer unit from continuously generating heat and rising in temperature, further improving the transfer quality.

According to an embodiment of the invention, the duty ratio of the unipolar electric signal is 1% to 99%, or 30% to 60%. The unipolar electrical signal of this embodiment provides an adjustable duty cycle range.

According to one embodiment of the invention, the biological transfer printing device further comprises a temperature sensor for measuring the temperature information of the first flat electrode layer and/or the second electrode layer in real time during the transfer printing process;

wherein the temperature information is fed back to the control unit, and the control unit is further configured to: dynamically adjusting the duty cycle during transfer based at least on the temperature information. The biological transfer printing device of the embodiment can dynamically adjust the duty ratio based on temperature information feedback, the adjustment of the unipolar electric signals is more accurate, the maximization of transfer printing work doing efficiency is more favorably realized, the temperatures of a gel layer, a carrier film and the like are further reduced, the heat emission is reduced, and the transfer printing quality is improved.

The bio-transfer device according to an embodiment of the invention, wherein the control unit is further configured to: adjusting at least the duty ratio and/or the maximum instantaneous power to make the temperature of the transfer unit lower than 60-70 ℃ in the transfer process. Therefore, the occurrence of scorching of the gel layer and the like of the transfer process transfer unit can be completely avoided.

According to an embodiment of the present invention, the bio-transfer device, wherein the transfer unit includes:

the first flat electrode layer and the second flat electrode layer are arranged in parallel; and

the first buffer dielectric layer is positioned between the first flat electrode layer and the second flat electrode layer, and the first buffer dielectric layer comprises a gel layer containing biological macromolecules, a bearing film and a second buffer dielectric layer;

wherein the biomacromolecules in the gel layer are electrophoresed to the carrier film under the action of the applied unipolar electric signal.

According to an embodiment of the present invention, a ratio of the resistances of the first buffer medium layer, the second buffer medium layer and the carrier film to the resistance of the gel layer is less than or equal to 3. The transfer unit is arranged, so that transfer acting efficiency is further improved.

According to one embodiment of the invention, the control unit and at least one transfer unit realize energy and/or information transmission through non-contact electromagnetic coupling. Therefore, the structure of the transfer printing device in the embodiment of the invention can be designed to be more compact, the whole water resistance, biological or chemical pollution resistance can be realized more easily, the use is more convenient and flexible, and the transfer printing device is very suitable for being applied to a biological laboratory.

According to an embodiment of the present invention, the bio-transfer device, wherein the control unit includes:

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

a unipolar electric signal generation module for generating the unipolar electric signal according to the adjusted control parameter;

wherein the adjusted control parameter comprises at least one of a waveform, a frequency, a voltage value, a current value, and a duty cycle of the unipolar electric signal;

wherein the measurement information is temperature information of the first flat electrode layer and/or the second flat electrode layer measured in real time during the transfer process, or timing information including at least one of a waveform, a frequency, a voltage value, a current value, and a duty ratio of the unipolar electric signal, and temperature information of the first flat electrode layer and/or the second flat electrode layer, which is recorded during the transfer process. The biological transfer printing device provided by the embodiment of the invention is beneficial to further reducing the work consumed by heat radiation and temperature increase in the transfer printing process, thereby being beneficial to reducing the temperature of the transfer unit and improving the transfer printing quality.

According to a second aspect of the present invention, there is provided a biological transfer system comprising:

a plurality of the above-described bio-transfer devices;

a cloud server coupled to control units of a plurality of the bio-transfer devices;

wherein the cloud server is configured to include:

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

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

the sending module is used for sending the control parameters to the corresponding control unit of the biological transfer printing device;

wherein the control parameter comprises at least one of a waveform, a frequency, a voltage value, a current value, and a duty cycle of the unipolar electric signal; the measurement information is temperature information of the first flat electrode layer and/or the second flat electrode layer measured in real time during the transfer process, or timing information including at least one of a waveform, a frequency, a voltage value, a current value, and a duty ratio of the unipolar electric signal, and temperature information of the first flat electrode layer and/or the second flat electrode layer, which is recorded during the transfer process.

The biological transfer printing system can quickly determine the control parameters which accord with the current transfer printing process based on the historical data information, is easy to realize the self-learning function, greatly improves the user experience, and greatly reduces the professional skill and experience requirements of users or experimenters.

According to a third aspect of the present invention, there is provided a control method for a biological transfer apparatus including one or more transfer units including a first flat electrode layer and a second flat electrode layer, and a gel layer and a carrier film between the first flat electrode layer and the second flat electrode layer; and uniformly applying a unipolar electric signal which periodically fluctuates according to a preset amplitude to the first flat electrode layer and the second flat electrode layer of one or more transfer units, wherein the ratio of the instantaneous power corresponding to the highest point of the preset amplitude to the instantaneous power corresponding to the lowest point of the preset amplitude is greater than or equal to 2.

The control method of the biological transfer printing device is beneficial to realizing maximization of transfer printing work efficiency, ensures the transfer efficiency, reduces the temperature of a transfer unit in the transfer printing process, reduces heat generation and improves the transfer printing quality.

According to an embodiment of the present invention, the fluctuating unipolar electric signal includes a square wave voltage signal with an adjustable duty ratio, or at least one of an intermittent sine wave signal, a triangular wave signal, a sawtooth wave signal, and a staircase signal. The unipolar electric signal of this embodiment is easy to generate and the relevant parameters are easy to control.

According to an embodiment of the invention, the fluctuation frequency of the unipolar electric signal is greater than or equal to 1Hz and less than or equal to 100Hz, or greater than or equal to 5Hz and less than or equal to 20Hz. In the frequency range of this embodiment, it is more favorable to avoid the transfer unit from continuously generating heat and rising in temperature, further improving the transfer quality.

The control method according to an embodiment of the present invention, wherein the control method further includes:

generating adjusted control parameters based on the measurement information or/and the transfer quality information, the adjusted control parameters including at least one of a waveform, a frequency, a voltage value, a current value, and a duty ratio of the unipolar electric signal; and

updating the unipolar electric signal according to the adjusted control parameter, and applying the updated unipolar electric signal to the transfer unit. The control method of the biological transfer printing device can dynamically adjust the duty ratio based on the temperature information feedback, the adjustment of the unipolar electric signal is more accurate, the maximization of the transfer printing work doing efficiency is more favorably realized, the temperatures of the gel layer, the bearing film and the like are further reduced, the heat emission is reduced, and the transfer printing quality is improved.

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

Drawings

The above and other objects and advantages of the present invention will become more apparent from the following detailed description when taken in conjunction with the accompanying drawings, in which like or similar elements are designated by like reference numerals.

Fig. 1 is a schematic configuration diagram of a biological transfer apparatus according to a first embodiment of the present invention.

Fig. 2 is a schematic structural view of a transfer unit in the biotransfer apparatus of the first embodiment shown in fig. 1.

Fig. 3 is a schematic diagram of a unipolar electric signal according to an embodiment of the present invention, in which fig. 3 (a) is a square wave voltage signal with an adjustable duty ratio, and fig. 3 (b) is a transformation example of fig. 3 (a).

Fig. 4 is a schematic diagram of a unipolar electric signal according to still another embodiment of the present invention, in which fig. 4 (a) and 4 (b) are sinusoidal voltage signals, fig. 4 (c) and 4 (d) are triangular voltage signals, fig. 4 (e) and 4 (f) are sawtooth voltage signals, and fig. 4 (g) and 4 (h) are stepped voltage signals.

FIG. 5 is a schematic diagram of the functional block configuration of the control unit of the bio-transfer device according to an embodiment of the present invention.

Fig. 6 is a schematic diagram showing a relationship between the resistance ratio and the transfer effective work.

Fig. 7 is a calculated transfer effective power distribution diagram.

FIG. 8 is a schematic view of the control principle of the bio-transfer device according to the embodiment of the present invention.

Fig. 9 is a schematic structural view of a biological transfer apparatus according to a second embodiment of the present invention.

Fig. 10 is a schematic structural view of a biological transfer apparatus according to a third embodiment of the present invention.

Fig. 11 is a schematic diagram of a non-contact electromagnetically coupled power supply module for use in a biotransformation apparatus according to an embodiment of the present invention.

Fig. 12 is a schematic configuration diagram of a first embodiment of a bio-transfer system formed based on the bio-transfer device of the embodiment shown in fig. 9.

Fig. 13 is a schematic configuration diagram of a second embodiment of a bio-transfer system formed based on the bio-transfer device of the embodiment shown in fig. 10.

Fig. 14 is a schematic block diagram of a cloud server of the bio-transfer system according to an embodiment of the present invention.

Detailed Description

The present invention will now be described more fully with reference to the accompanying drawings, in which exemplary embodiments of the invention are shown. This invention may, however, be embodied in many different forms and should not be construed as limited to the embodiments set forth herein. Rather, these embodiments are provided so that this disclosure will be thorough and complete, and will fully convey the concept of the invention to those skilled in the art. In the drawings, the same reference numerals denote the same elements or components, and thus, their description will be omitted.

In the drawings, the sizes of layers and regions of the transfer unit are exaggerated for clarity, and the sizes of the respective layers in the transfer unit in the drawings do not limit the actual sizes and dimensional proportional relationships of the respective layers.

Some of the block diagrams shown in the figures are functional entities, which do not necessarily have to correspond to physically or logically separate entities, which may in some cases be implemented in software, or in yet other cases by one or more hardware modules or integrated circuits, or in yet other cases in different network and/or processor devices and/or microcontroller devices.

FIG. 1 is a schematic view of a configuration of a bio-transfer device according to an embodiment of the present invention. The bio-transfer 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 control unit 11 mainly functions to control a specific operation process of the transfer unit 130, specifically by controlling an electric signal applied to the transfer unit 130.

Fig. 2 is a schematic structural view of a transfer unit in the bio-transfer apparatus according to the embodiment shown in fig. 1. Referring to fig. 2, the transfer unit 130 is mainly formed of upper and lower electrodes and a stacked structure 131 stacked therebetween, and specifically includes an upper plate electrode layer 1301 and a lower plate electrode layer 1302, and an upper buffer dielectric layer 131e between the upper plate electrode layer 1301 and the lower plate electrode layer 1302, a gel layer 131d containing bio-macromolecules, a carrier film 131c, and a lower buffer dielectric layer 131b. The function of the transfer unit 130 is to apply electrical signals to the upper plate electrode layer 1301 and the lower plate electrode layer 1302 so as to generate an electric field E similar to the direction indicated by the dotted arrow in fig. 2, and the bio-macromolecules in the gel layer 131d are electrophoretically moved under the action of the electric field E in the direction indicated by the time-limit arrow in fig. 2, and then transferred onto the carrier film 131c, thereby completing the transfer process.

In the following embodiments of the present invention, a protein transfer process will be exemplified, however, it should be understood that the bio-macromolecule to be transferred in the gel layer 131d is not limited to protein, but may be other similar bio-macromolecules such as DNA. When the biological transfer device 10 is used for transferring proteins, it is specifically a western blotting device; when the bio-transfer device 10 is used to transfer 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 is not limited. In the present invention, the "flat electrode layer" refers to a planar single layered electrode structure (for each stacked structure 131 of the transfer unit), when a certain electric signal is applied to the upper flat electrode layer 1301 and the lower flat electrode layer 1302, the same electric signal is uniformly applied between the upper flat electrode layer 1301 and the lower flat electrode layer 1302, a uniform electric field E as illustrated in fig. 2 is formed between the upper flat electrode layer 1301 and the lower flat electrode layer 1302, the electric field E is uniformly applied to different sized proteins in the gel layer 131d, that is, the different sized proteins are electrophoretically moved by the uniform electric field E. The upper flat electrode layer 1301 and the lower flat electrode layer 1302 of the embodiment of the invention are simple in structure and preparation.

The term "uniformly" applying the electric field means that the electric fields for causing the electrophoretic motion of the biomolecules in the gel layer 131d act uniformly and simultaneously with respect to all the biomolecules such as proteins in the gel layer 131 d.

It should be noted that the principle of applying an electric signal to a Protein by the upper plate electrode layer 1301 and the lower plate electrode layer 1302 according to the embodiment of the present invention is completely different from the principle of applying an electric field disclosed in the patent No. US8721860B2 entitled "Protein Multi-blocking Method and Device". In US8721860B2, different electrical signals are controlled to be applied to different electrode layers, so that the same electrical signal cannot be uniformly applied to different electrode layers of the composite electrode layer, and for different sized proteins, the same electrical signal is affected by different sized electric fields to perform electrophoretic movement, and different electrical signals applied to different electrode layers can generate different transfer efficiencies for different sized proteins respectively. Therefore, in US8721860B2, the electrical signals are not applied to the upper and lower electrode layers in a spatially uniform manner.

Continuing with FIG. 2, in one embodiment, the upper plate electrode layer 1301 and the lower plate electrode layer 1302 are disposed in parallel, such that not only can an electrical signal be applied between the upper plate electrode layer 1301 and the lower plate electrode layer 1302 in a spatially uniform manner, but also the electric field E generated by the electrical signal is distributed substantially uniformly in the left-right direction as shown in FIG. 2.

Continuing with FIG. 2, in one embodiment, the upper buffer dielectric layer 131e is disposed between the gel layer 131d and the upper plate electrode layer 1301, and the lower buffer dielectric layer 131b is disposed between the carrier film 131c and the lower plate electrode layer 1302, which may be formed of a material with buffer gel or filter paper, which has an electrically conductive function, and also has a protective function for the gel layer 131d and the carrier film 131c, and provides buffer ions (buffer ions) for transfer. The resistances of the upper and lower buffer dielectric layers 131e and 131b, which may be measured or estimated according to the characteristics of the materials selectively used, are recorded as R, respectively _e And R _b . Note that the resistance R of the upper buffer dielectric layer 131e _e And/or R of lower buffer dielectric layer 131b _b It varies according to the material, the amount of buffer and the temperature change.

Continuing with fig. 2, in one embodiment, the gel layer 131d may be a biomass membrane, such as a gel having a gradient concentration of 4% -20% or 4% -12%, or a uniform concentration of gel in the range of 4% -12%, where different sized proteins to be transferred may be electrophoresed in the gel layer 131 d; the thickness of the gel layer 131d may range from 0.5mm to 2.0mm, and may be 1mm, for example; the resistance of the gel layer 131d is measured or estimated from the properties of the material selected for use, and is recorded as R _d . The support membrane 131c may be, for example, a nitrocellulose membrane or a PVDF (polyvinylidene fluoride) membrane, a specific material thereofThe resistance of the carrier film 131c, which is recorded as R, is measured or estimated according to the characteristics of the material selected for use, and is not limited thereto _c . The resistance R of the gel layer 131d _d And/or R of the carrier film 131c _c It varies according to the material, the amount of buffer and the temperature change.

The applicant should point out that the layers of the laminated structure 131 of the transfer unit 130, particularly the gel layer 131d, are liable to be denatured at high temperature, and therefore, it is necessary to control the temperature of the transfer unit 130 (particularly the gel layer 131 d) during transfer, for example, below 60 ℃ to 70 ℃ to ensure the transfer quality. However, in the related art, the transfer operation is performed with a constant voltage, a step-increasing voltage, or a constant current signal applied to the transfer unit 130; the applicant has found that as the transfer process proceeds or ends, its temperature tends to exceed 60-70 ℃, for example, in the case of a constant voltage signal of 25V applied for 10min, the glue in the transfer unit 130 may even be burnt; further, in order to increase the transfer rate of large proteins, the transfer efficiency is not necessarily improved by applying a high voltage or a high current, and the temperature of the transfer unit 130 is easily increased, so that the transfer quality is difficult to be ensured.

Continuing with fig. 1, the control unit 11 in the transfer device 10 is configured to apply an undulating unipolar electrical signal, the unipolar electrical signal of the present invention being periodically undulating by a predetermined amplitude, to the upper and lower flat electrode layers 1301, 1302 of one or more transfer units 130, wherein the 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 2. It should be noted that the specific magnitude of the predetermined amplitude may be constant or may vary, that is, in the case that the ratio of the instantaneous power at the highest point of the predetermined amplitude to the instantaneous power at the lowest point corresponding to the predetermined amplitude is greater than or equal to 2, during the continuous fluctuation of the signal period, the ratio may be constant or may vary, for example, relatively constant in one control period, and the signal period of each control period may be adjusted and varied between a plurality of control periods. The time interval during which the above defined heave course is completed once is defined as one signal period of the unipolar electric signal, e.g. the time interval during which the highest or lowest point of the above predetermined amplitude occurs continuously is defined as one signal period of the unipolar electric signal.

For example, if the unipolar electric signal is a 10Hz square wave signal, and the peak voltage is 25V and the valley voltage is 5V at the beginning of transfer, the signal cycle of the unipolar electric signal is 100 milliseconds, and the instantaneous power ratio of the extreme points in one signal cycle is 25 × 25/(5 × 5) =25; after the transfer operation is continued for, for example, 1 second, the control unit adjusts the control parameter, the unipolar electric signal is still a square wave signal, but the signal frequency of the unipolar electric signal is changed to 20Hz, the peak voltage is changed to 20V, and the valley voltage is changed to 5V, then the signal period of the unipolar electric signal corresponds to 50 milliseconds, and the instantaneous power ratio of an 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, further obtains different square wave signals, and thus continuously controls or adjusts the unipolar electric signal at a predetermined time interval (for example, 1 second as described above) during the transfer operation, and then the predetermined time interval (for example, 1 second) is the control period referred to in this application. The principle of adjusting the unipolar electric 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 power within the signal period; the ratio of the maximum instantaneous power to the minimum instantaneous power within each signal period of the unipolar electrical signal is greater than or equal to about 2, which in one embodiment is greater than or equal to, for example, 2.5. It is to be understood that in each signal cycle the unipolar electric signal defines the amplitude of the undulations in such a way that the ratio of the maximum instantaneous power to the minimum instantaneous power in each signal cycle is greater than or equal to 2, the maximum instantaneous power corresponding to a high point or peak of the waveform (i.e. the highest point of said predetermined amplitude) and the minimum instantaneous power corresponding to a low point or trough of the waveform (the lowest point of said predetermined amplitude).

Specifically, the unipolar electric signal may be a voltage signal or a current signal, and may be classified according to waveforms, and specifically, but not limited to, the unipolar electric signal may be a square wave signal, a sine wave signal, a sawtooth wave signal, or a step signal, or a combination thereof; the unipolar electrical signal may be continuous or intermittent. The waveform, frequency, voltage value, current value, duty ratio, and the like of the unipolar electric signal are important characteristics or parameters reflecting the unipolar electric signal. Illustratively, the unipolar electric signal is specifically a square wave voltage signal with an adjustable duty ratio, the square wave voltage signal is relatively convenient to control and easy to generate, and the adjustment of the duty ratio of the square wave voltage signal can be realized by adjusting the pulse width.

Fig. 3 is a schematic diagram of a unipolar electric signal according to an embodiment of the present invention, in which fig. 3 (a) is a square wave voltage signal 91 with an adjustable duty cycle, and fig. 3 (b) is a conversion example of fig. 3 (a). As shown in FIG. 3 (a), the voltage peak (i.e., high level) of the square wave voltage signal 91 is V _p The low level is 0V, V _p Greater than or equal to 1V and less than or equal to 30V (e.g., 20V), corresponding to V _p The peak current generated at the transfer unit 130 is greater than or equal to 0.1A and less than or equal to 10A (e.g., 6A); corresponding V of each signal period _p A maximum instantaneous power can be generated and a minimum instantaneous power can be generated at a corresponding low level of each signal cycle. The signal period T of the square wave voltage signal 91 can be set adjustably even in the same transfer process, and particularly, the signal period T of the square wave voltage signal 91 can be controlled by controlling the frequency magnitude thereof; in an embodiment, the frequency of the square wave voltage signal 91 is greater than or equal to 1Hz and less than or equal to 100Hz, or for example greater than or equal to 5Hz and less than or equal to 20Hz, for example 10Hz. The duty cycle of the square wave voltage signal 91 is also adjustable, with the duty cycle ranging from 1% to 99%, or for example 30% to 60%.

As shown in fig. 3 (b), the square wave voltage signal 91 'is an alternative embodiment of the square wave voltage signal 91, and in the square wave voltage signal 91', the voltage peak value, the low level size, the signal period T and/or the duty ratio, etc. may be varied. By setting 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 2. It should be understood that the ratio of the maximum instantaneous power to the minimum instantaneous power in each signal period is not necessarily substantially constant.

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

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

As shown in FIG. 4 (c), the signal 93 is a triangular wave voltage signal, which is an intermittent signal corresponding to the voltage peak V _p A maximum instantaneous power per signal period may be generated and a minimum instantaneous power may be generated at a level corresponding to 0. As shown in fig. 4 (d), the signal 93' is also a triangular wave voltage signal, but it is a continuous electrical signal corresponding to the voltage peak V _p A maximum instantaneous power per signal period may be generated and a minimum instantaneous power per signal period may be generated at a corresponding minimum voltage (which is not limited to 0).

As shown in FIGS. 4 (e) and (f), the signals 94 and 94' are both sawtooth voltage signals, which are intermittent signals corresponding to voltage peaks V _p A maximum instantaneous power per signal period may be generated and a minimum instantaneous power may be generated at a level corresponding to 0.

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 step manner when the voltage waveform changes in a fluctuating manner, and the number of step changes in each signal period and the voltage magnitude of each step change are not limited; corresponding voltage peak value V _p A maximum instantaneous power per signal period may be generated and a minimum instantaneous power may be generated at a level corresponding to 0. As shown in FIG. 4 (h), signal 95' is also a step voltage signal, but at a low level greater than 0V, corresponding to a voltage peak V _p The maximum instantaneous power per signal period may be generated and the minimum instantaneous power per signal period may be generated at the corresponding minimum voltage.

In the above unipolar electric signals of the embodiment shown in fig. 4, they are of periodic nature; for voltage signals, and for the voltage in each signal period, by setting the voltage peak V in each signal period _p And the minimum voltage, the ratio of the maximum instantaneous power to the minimum instantaneous power in each signal period can be realized to be greater than or equal to 2; for the current signal, by setting the current peak value and the minimum current value in each signal period, the ratio of the maximum instantaneous power to the minimum instantaneous power in each signal period can be realized to be greater than or equal to 2. It is to be understood that the specific waveform of the undulating unipolar electric signal is not limited to the above embodiments, which may be specifically selected to be arranged according to the needs of a specific application. For unipolar electrical signals that vary in a wavy manner, for example sine waves, triangular waves, sawtooth waves or steps, the duty cycles can be defined for them individually, for example, where the average of the maximum voltage and the minimum voltage in each signal cycle corresponds, the ratio of the time period greater than this average to this signal cycle T is defined as the duty cycle; thus, also like the square wave voltage signal 91, their duty cycles are also adjustably set, for example in the range of 1% -99%, or for example 30% -60%.

Continuing with fig. 1, control unit 11 is for generating the voltage signal of the above embodiment (exemplified by generating square wave voltage signal 91 as in fig. 3 (a)), which specifically includes waveform generator 110, controller 150, power supply module 190, human Machine Interface (HMI) 170, real time clock 160, voltage and/or current detection component 120, and the like. Among them, the controller 150 is a core component of the control unit 11, and is capable of outputting control parameters to the waveform generator 110, so as to generate corresponding unipolar electric signals, such as square wave voltage signals; the controller 150 may have measurement, calculation and control or even memory functions, the specific working principle of which will be disclosed in detail later.

Wherein the voltage and current detecting part 120 serves to detect the voltage U and/or the current I applied to the transferring unit 130 in real time, and feed it back to the controller 150 as measurement information,

the power module 190 is used to provide power to the bio-transfer device 10, for example, it can provide ac power or dc power to the waveform generator 110 to generate unipolar electric signals of corresponding waveforms, and can also provide low voltage dc power to the controller 150, and can supply power to the real-time clock 160, the voltage and current detection part 120, and the like electrically connected thereto through the controller 150.

The real-time clock 160 may provide the current actual time to the waveform generator 110 and the controller 150, and based on the actual time information, may control to generate a square wave signal with parameters such as corresponding frequency (or signal period), duty ratio, etc.; the measurement information received by the controller 150 or information measured by itself, such as voltage or current information fed back by the voltage and current detecting part 120, is combined with actual time information provided by the real-time clock 160 to generate measurement information having a corresponding time stamp, so that timing information corresponding to the measurement information can be obtained, which will be used in the calculation process of the control algorithm. In particular, the real time clock 160 may be provided embedded in the controller 150.

Among other things, the human-machine interface 170 is used to implement interaction with a user, for example, a function of selecting or setting a transfer option, a transfer parameter, and a function of starting or stopping a transfer process by the user, and a function of providing a user feedback to the controller 150, and a function of presenting status information, etc. to the user during the transfer process. The human-machine interface 170 is not limited to be integrally mounted on the bio-transfer device 10, and may be provided 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, through which communication unit 180, the control unit 11 is enabled to be connected to an external smart terminal (e.g., a tablet computer, a smart phone, etc.) and/or a cloud computing server, etc., and a part of 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 computer (IPAD), etc.

As further shown in fig. 1, in one embodiment, the temperature sensor 140 is further disposed in the bio-transfer device 10 corresponding to the transfer unit 130, and 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, and the measured temperature information substantially accurately reflects the temperature of the current gel layer 131 d. The temperature information measured by the temperature sensor 140 may be fed back as measurement information to the control unit 11, specifically, for example, to the controller 150. The temperature sensor 140 may in particular be provided integrally with the transfer unit 130.

FIG. 5 is a schematic structural diagram of functional modules of a control unit 11 of a biological transfer apparatus according to an embodiment of the present invention. As shown in fig. 5 and fig. 1, in an embodiment, the controller 150 controls the waveform generator 110 by outputting a control parameter, and therefore, the control unit 11 is provided with a control parameter generating module 151, which may be provided in the controller 150 or implemented by the controller 150; the control unit 11 is further provided with a unipolar electric 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 electric signal generating module 111 is configured to generate a unipolar electric signal (e.g., a square wave voltage signal) according to the adjusted control parameter.

Wherein the control parameter comprises at least one of a waveform, a frequency, a voltage value, a current value, and a duty cycle of the unipolar electric signal; the measurement information is temperature information of the upper plate electrode layer 1301 and/or the lower plate electrode layer 1302 measured in real time during the transfer process, or timing information of at least one of a waveform (for example, a square wave, a sawtooth wave, a triangular wave, a sine wave, or the like) including a unipolar electric signal, a voltage value U, a current value I, and a duty ratio, and temperature information of the upper plate electrode layer 1301 and/or the lower plate electrode layer 1302 recorded during the transfer process. Therefore, the method can realize real-time adjustment of parameters such as the waveform, the duty ratio, the voltage value, the current value and the like of the unipolar electric signal according to the feedback of the measurement information or/and the transfer quality information, is beneficial to realizing the maximization of the transfer work efficiency, reduces the temperature of a gel layer, a bearing film and the like, reduces the heat emission and improves the transfer quality while ensuring the transfer efficiency.

In an embodiment, the control unit 11 further comprises an input module 154 for receiving input transfer quality information, illustratively, at the end of a transfer process for a certain transfer unit 130, manually evaluating the quality of the results obtained for the transfer process and inputting feedback, e.g. from the human machine interface 170 or an external device connected to the communication unit 180, the input module 154 receiving the transfer quality information corresponding to each transfer process. The input module 154 may be provided in the controller 150 or implemented by the controller 150.

In evaluating the transfer quality information per transfer process, the user may evaluate according to a predetermined index or criterion, specifically may evaluate to obtain the transfer quality information expressed by a grade or a score, and exemplarily determine the transfer quality information Qt based on the transfer quality scoring criterion of the following table one.

Watch 1

Evaluation criteria	Weight of	Transfer quality scoring
			Transfer efficiency of print	50％	0-5
Gel layer condition	20％	0-2
			Condition of carrier film	20％	0-2
Morphology of the print	5％	0-0.5
			Buffer dielectric layer condition	5％	0-0.5
Transfer quality information (Qt)	100％	0-10

The factors for evaluating the transfer quality information include, but are not limited to, transfer efficiency of the print, a condition of the gel layer, a condition of the carrier film, a form of the print, a condition of the buffer medium layer, and the like, and each factor is given a corresponding weight. For the transfer efficiency of the print, the score can be determined by reading print (Marker) information (e.g., the number of prints remaining in the gel layer 131 d); for gel layer condition, it can be scored according to whether its condition corresponds to scorch (burned), curl (curved), good, etc.; for the status of the carrier film, the rating can be made according to whether the status corresponds to coking, curling, good, etc.; for the morphology of the print, it can be scored according to whether its condition corresponds to diffusion, clarity, etc.; the condition of the buffer medium layer can be correspondingly graded according to whether the condition corresponds to good coking and the like. And finally, integrating all the transfer quality scores, and calculating to obtain the total transfer quality score, namely the transfer quality information Qt, based on 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 may 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 determining the transfer quality information according to the above-exemplified transfer quality scoring criterion, and may specifically be set according to actual applications so as to help the user to obtain the transfer quality information as truly as possible.

In one embodiment, the control unit 11 further comprises a storage module 153 for recording the measurement information or/and the transfer quality information obtained from each transfer process. The storage module 153 may be provided in the controller 150 or implemented by the controller 150. The measurement information may be various information measured in real time by the transfer process, such as 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 may be called by the control parameter generation module 151, and of course, may be selectively transmitted to an external device connected thereto through the communication unit 180.

In an embodiment, the storage module 153 may also store corresponding control parameters, for example, generated by the control parameter generation module 151, corresponding to the transfer unit 130. When the transfer unit 130 of a certain transfer process corresponds to the same transfer unit 130 of a certain history transfer process, the control parameter generation module 151 may directly call the control parameter stored in the storage module 153 corresponding to the certain history transfer process, so as to facilitate rapid completion of the experimental design (for example, including setting of the parameter) of the certain transfer process, and the generated unipolar electric signal with fluctuation variation will also be applicable to the certain transfer process, thereby facilitating rapid and high-quality completion of the transfer process.

The bio-transfer device 10 of the embodiment of the present invention can realize that the periodically fluctuating unipolar electric signal is applied to the transfer unit 130, and the ratio of the maximum instantaneous power to the minimum instantaneous power in each signal period of the unipolar electric signal is greater than or equal to about 2; in the peak time period or time point corresponding to the maximum instantaneous power, since there is a valley time period or time point with smaller power of voltage or current drop in the subsequent time, the transfer unit 130 does not continuously operate in a high power state, and thus a situation of continuous overheat temperature rise similar to a constant current or constant voltage electrical signal does not occur, and thus, the magnitude of the voltage or current applied to the gel layer 131d of the transfer unit 130 may not be substantially limited; thus, on one hand, the larger instantaneous power at the peak time period or time point can effectively push the protein and other biological macromolecules (especially large mass) to do electrophoresis movement, thereby ensuring and even improving the transfer efficiency, on the other hand, the smaller instantaneous power at the trough time period or time point can effectively prevent the temperature of the transfer unit 130 from continuously rising due to ohmic heating, the resistance of the transfer unit 130 can not continuously increase due to heating, the temperature or resistance of the transfer unit 130 is reduced, and the work with smaller proportion (namely W) in the work at the peak time period or time point is also enabled to be _Tm ) Heat radiation and temperature increase for the transfer unit 130, a larger proportion of work (i.e. transfer effective work W) _em ) Used for driving protein and other biological macromolecules to do electrophoresis movement. That is, in a signal cycle or a plurality of signal cycles of the unipolar electric signal that fluctuates and changes, the arrangement of the trough time period or the time point of the smaller power is favorable for improving the transfer printing work efficiency of the peak time period or the time point of the larger power, so that the transfer printing work efficiency of the unipolar electric signal is integrally improved, and the transfer efficiency can be ensured even ensuredThe temperature of the transfer unit 130 is slowly increased in the transfer process, and the transfer quality is good.

The above-described technical effects of the biological transfer apparatus 10 according to the embodiment of the present invention will be described below by comparison tests.

In the comparison test, the transfer effect comparison was performed based on the square wave voltage signal and the constant voltage signal generated by the bio-transfer device 10 of the embodiment shown in fig. 1, in which the high level of the square wave voltage signal (i.e., the peak voltage Vp) is the same as the voltage of the constant voltage signal and the time of the transfer operation is the same, and the transfer unit 130 to which the square wave voltage signal and the constant voltage signal are applied is the same. It should be noted that, in the process of applying a constant voltage signal to perform a transfer operation, it is necessary to perform current limiting in accordance with the voltage signal, or an excessive current is likely to be generated to burn the gel layer, the carrier film, and the like.

In the first comparison tests, each test simultaneously performed the transfer operation of 2 gel layers 131d juxtaposed in the transfer unit 130, and the specific parameters of the two tests for comparison are referred to the following table two.

Watch 2

When the duty ratio of the applied square wave voltage signal is 40%, corresponding to a high level, the peak value of the measured current is 6A at the initial stage of the transfer process, and the peak value of the measured current is 2A at the end stage of the transfer process, wherein the peak value is equivalently 4A; thus, 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 the temperature sensor 140) is about 65 ℃. In the above calculation process, since the current is very small and the average power is also very small for the low level 5V, the power for generating heat is very small for the high level stage, and therefore, the process of calculating the total power consumption for generating heat is neglected.

In comparison, in the transfer process corresponding to the constant voltage signal, since the current is limited to 2.5A, the applied actual voltage measured at the initial stage of the transfer process is 18V, and the applied actual voltage measured at the end stage of the transfer process is 24V; equivalently taking the median value of 21V; thus, 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 the temperature sensor 140) is about 78 ℃.

In the second alignment test, each test simultaneously performed the transfer operation of the 4-gel layer 131d juxtaposed in the transfer unit 130, and specific parameters of the two tests for alignment are shown in table three below

Watch III

When the duty ratio of the applied square wave voltage signal is 40%, and corresponds to a high level, the measured current peak value is 12A at the initial stage of the transfer process, and the measured current peak value is 6A at the end stage of the transfer process, and the current peak value is equivalently 4A; thus, 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 the temperature sensor 140) is about 72 ℃. In the above calculation process, since the current is very small and the average power is also very small for the low level 5V, the power for generating heat is very small for the high level stage, and therefore, the process of calculating the total power consumption for generating heat is neglected.

In comparison, in the transfer process corresponding to the constant voltage signal, since the current is limited to 5A, the measured applied actual voltage is 15V at the initial stage of the transfer process, and the measured applied actual voltage is 25V at the end stage of the transfer process; equivalently taking the median value of 20V; thus, the average power during transfer 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 the temperature sensor 140) is about 86 ℃.

From the above first and second comparison tests, it can be found that less work is converted into heat during the transfer process corresponding to the square wave voltage signal, and the transfer operation can be completed as well, the gel layer temperature is also lower, and 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 to achieve electrophoretic motion of the protein during the transfer process, i.e., the use of square wave voltage signals has a higher transfer work efficiency than constant voltage signals.

The following specific example theory explains why the transfer operation using the square wave voltage signal has relatively higher transfer work efficiency.

I. First, assuming that a voltage applied to the stacked structure (including the upper buffer dielectric layer 131e, the gel layer 131d, the carrier film 131c, and the lower buffer dielectric layer 131 b) of the transfer unit 130 is U, the thicknesses of the upper buffer dielectric layer 131e, the gel layer 131d, the carrier film 131c, and the lower buffer dielectric layer 131b are d, respectively _e 、d _d 、d _c 、d _b The resistivities are respectively rho and rho _d 、ρ _c And ρ _b The area (cross-sectional area perpendicular to the electric field E) is S, and the mobility of the target bio-macromolecule in the gel layer 131d and the carrier film 131c is m _d And m _c . The above voltage U, thickness d, and area S may be regarded as constant values during transfer, but their resistivity ρ _e 、ρ _d 、ρ _c And ρ _b Is variable during the transfer process. The change in resistivity is dependent on the material properties and temperature, and the change is due to a change in temperature and/or material composition. Mobility m _d And m _c Depending on the size and shape of the target bio-macromolecules, the structure (e.g., pore size) and materials of the gel layer 131d and the carrier film 131c, and the chemical and physical characteristics of the upper and lower buffer dielectric layers 131e and 131b, among others.

II, resistance R of the gel layer 131d, the carrier film 131c, the upper buffer dielectric layer 131e and the lower buffer dielectric layer 131b _d 、R _c 、R _b And R _e Can be controlled byAre represented by the formulae (1-1-1), (1-1-2), (1-1-3) and (1-1-4), respectively:

wherein R is _d 、R _c 、R _b And R _e Respectively, the resistances of the gel layer 131d, the carrier film 131c, the upper buffer dielectric layer 131e, and the lower buffer dielectric layer 131b.

Further, according to ohm's law, the voltage U applied to the gel layer 131d _d And electric field intensity E _d Can be represented by the following relations (1-2-1) and (1-2-2), respectively:

wherein, the voltage U _d For applying a voltage to the gel layer 131d, E _d For applying an electric field strength to the gel layer 131 d.

Further, the transfer velocity v of the target biomacromolecule is further represented based on the following relational expression (1-3):

where m is the mobility of the biological macromolecule.

V. hypothesis, m _d ＞＞m _c Thus, m _c Can be ignored during the transfer time t, then the transfer work effective W of the target biomacromolecule _em Can be calculated based on the following relations (1-4):

wherein, W _em For transferring the effective work, the larger the transfer effective work proportion is, the larger the transfer acting efficiency is; q is the charge of the target biological macromolecule, and 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 And R _e Is typically in the range of 5-20 ohms, and the transfer effective work W can be determined substantially from the above relations (1-4) _em (see ordinate) may be compared to the resistance (R) _c +R _b +R _e )/R _d (see abscissa) the relationship between them is schematically shown in fig. 6. Therefore, the resistance ratio (R) _c +R _b +R _e ) the/Rd can be controlled to be less than or equal to 3 to increase the transfer effective work W _em And further improve the efficiency of transfer work.

Taking the square wave voltage signal as an example, assuming that its duty cycle is μ, it may be set in the range of 1% to 99%, and further the relation (1-5) may be obtained based on the above relation (1-4):

VII. resistance R due to the upper and lower buffer dielectric layers 131e and 131b _b And R _e Obviously depending on the temperature T, which will generate power consumption for heat generation; the resistances of the upper and lower buffer dielectric layers 131e and 131b at the temperature T may be expressed by the following relations (1-6-1) and (1-6-2), respectivelyRespectively represent:

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 is _b (T) and R _b (T) represents the resistance of the upper buffer dielectric layer 131e and the lower buffer dielectric layer 131b at the temperature T, respectively; k is the heating coefficient (which can be estimated to be 10, for example); η duty threshold, which represents that the temperature of the transfer process does not increase any more if the duty of the square wave voltage signal is equal to the duty threshold, for example, it may be equal to 20%; r _b And R _e Which are the resistances of the upper buffer dielectric layer 131e and the lower buffer dielectric layer 131b before the transfer starts, respectively.

Based on the above relationships (1-5), (1-6-1) and (1-6-2), and assuming R _c ＜＜R _b R in the transfer process _d Is much less than R _b And R _e Can obtain the following effective work W for transfer _em The relational expression (1) to (7):

wherein the values k =10, η =20%, and the resistance R _d ，R _c ，R _b And R _e Is typically in the range of 5-20 ohms, a transfer effective power distribution map as shown in fig. 7 can be obtained through simulation calculation based on the above relations (1-7), wherein the higher the gradation, the higher the transfer effective power W is represented _em The smaller the size, the larger the opposite.

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

Further, as can be seen from fig. 7, in most cases, the effective work W is transferred when the duty ratio μ is equal to about 50% _em Is a watershed, namely the transfer effective work or the transfer work efficiency relatively reaches the peak value; for use in numerical simulationsThe 50% duty ratio of the square wave signal (such as the square wave signal shown in fig. 3 (a)) corresponds to 50% of full power output (the full power output corresponds to the power output when the duty ratio is 100%), that is, for unipolar electric signals of other waveforms, it is necessary to adjust the duty ratio within a range of 1% to 99% to achieve 50% or less (including 50%) of full power output. If the maximum instantaneous power P within the signal period _max With minimum instantaneous power P _min With a ratio less than 2, assuming that the power output P is calculated equivalently based on the following relationships (1-8):

P＝μP _max +(1-μ)P _min (1-8)

even if the duty ratio μ =0%, the regulated power output P cannot be adjusted to a full power output of 50% or less (including 50%). To this end, in an embodiment of the present invention, the maximum instantaneous power P within a signal period of a unipolar periodic signal _max With minimum instantaneous power P _min The ratio of the maximum power to the minimum power is greater than or equal to about 2, that is, the unipolar electrical signal fluctuates periodically by a predetermined amplitude, and the ratio of the instantaneous power at the maximum point of the predetermined amplitude to the instantaneous power at the minimum point of the defined amplitude is greater than or equal to 2; in this way, by adjusting the duty ratio μ to be 50% or less of the full power output, the transfer effective work or the transfer work efficiency can be relatively peaked.

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

Can realize large instantaneous power P _max With minimum instantaneous power P _min A ratio of greater than or equal to about 2; if the unipolar electric signal is current information, setting the ratio of the maximum instantaneous current x to the minimum instantaneous current in the signal period to be greater than or equal to about

Can realize large instantaneous power P _max To and fromSmall instantaneous power P _min The ratio of (a) to (b) is greater than or equal to about 2.

Fig. 8 further illustrates the control principle of the bio-transfer device, and from this point of view, the technical effects possessed by the bio-transfer device 10 of the above embodiment of the present invention can also be exemplarily explained. It is illustrated how the duty ratio μ generated by the control parameter generation module 151 of the controller 150 is adjusted based on the measurement information including the temperature information T of the transfer unit 130 measured in real time in the primary transfer process and how the control parameter is adjusted based on the transfer quality information Qt in the multiple transfer processes for the same transfer unit 130, thereby maximizing the transfer work efficiency. The square wave voltage signal is described as an example in conjunction with fig. 8.

As shown in fig. 8, in the first control loop process 191 of a certain transfer operation process, which corresponds to the mth transfer process of a certain transfer unit 130 (each transfer process may include a plurality of first control loop processes 191, i.e., a plurality of control periods), the transfer effective work W is realized by dynamically adjusting the duty ratio μ of each control period in real time _em And (4) maximizing.

First, effective work W is transferred _Em Calculated by the following relation (2-1):

W _em ＝W _m -W _Tm (2-1)；

wherein, W _m Representing the total work done by the square-wave voltage signal during transfer, W _Tm Indicating the work consumed for heat radiation and temperature increase during transfer.

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

where Δ t is the time interval of the first control loop process 191, which corresponds to one control period, one control period being greater than or equal to one signal period, e.g., which corresponds to a plurality of signal periods with unipolar electrical signals; m represents the m-th transfer process,U _m represents the voltage amplitude measured in the mth transfer process (if the minimum voltage of the unipolar electric signal in the control period is equal to 0, the voltage amplitude represents the voltage peak value), I _i Represents the measured current amplitude of the ith control period (if the minimum current in the control period is equal to 0, the current amplitude represents the peak value of the electric current), mu _i The duty cycle of the electrical signal representing the ith control period (the duty cycle of the unipolar electrical signal is constant during this control period, i.e. both are mu _i )，1≤i≤(n-1)。

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

wherein xi is _m Represents the equivalent emissivity of the transfer unit 130 in the mth transfer process; c represents the specific heat capacity of the transfer unit 130; t is a unit of _i And T _i-1 Respectively, the temperature information of the transfer unit 130 measured at the ith and (i-1) th control cycles.

Based on the above relational expressions (2-1), (2-2) and (2-3), the following relational expression (2-4) can be obtained:

thus, the effective work W is transferred _em Maximum, i.e. finding W _em The maximum value of the relative argument duty ratio μ can be expressed as the following relation (2-5):

based on the temperature information T and the current information I measured in the 1 st to (n-1) th control periods of the square wave voltage signal, etc., input to the above relational expression (2-5), the W in the 1 st to (n-1) th control periods is calculated _em The duty cycle corresponding to the maximum value of the duty cycle,the duty ratio is output as the duty ratio mu of the nth control period _n I.e. the duty cycle mu in the control parameter corresponding to the nth control period _n Waveform generator 110 is based on the duty cycle μ _n The duty ratio of the next control period (i.e., the nth control period) is adjusted. It is foreseen that the duty cycle μ is based on _n When the transfer operation is continued by the square wave voltage signal of the nth control period, W can be continuously caused to continue _em And (4) maximization.

During the second control loop 192, it is controlled based on the following relation (2-6):

wherein i represents the ith transfer process of the 1 st to (m-1) th transfer processes, m is an integer of 2 or more, Q _ti Indicating transfer quality information corresponding to the i-th transfer process, W _ei The transfer effective work of the i-th transfer process is expressed, which can be calculated by the above relational expression (2-5). Based on the previous (m-1) times and the mth time input transfer quality information, fitting calculation is carried out to obtain the equivalent thermal emissivity xi _m-1 Further, the equivalent emissivity is ξ _m-1 Equivalent voltage U as an intermediate input parameter in combination with the voltage measured for the (m-1) th secondary transfer process _m-1 Substituting the relation into the relation (2-4), and calculating to obtain the transfer effective work W of the mth transfer process _em The transfer effective work W _em Is the maximum value when the relative equivalent emissivity coefficient ξ is taken as the argument.

The equivalent emissivity xi obtained in the second control loop process 192 above can also be substituted into the relation (2-5) to obtain the corresponding duty ratio parameter in the first control loop process 191, so as to obtain the maximum transfer effective work.

The above first control cycle 191 and the second control cycle 192 are both for obtaining the transfer work efficiency as large as possible, thereby reducing the work consumed for heat radiation and temperature increase during the transfer process, contributing to lowering the temperature of the transfer unit 130, and improving 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 appreciated that the control parameter generation module 151 of the controller 150 is capable of automatically optimizing other control parameters than duty cycles, such as frequency or period, voltage values and/or current values, etc., the control parameter generation module 151 of the controller 150 has a self-learning function. This self-learning function may function in the following situations: for example, for a newly used bio-transfer apparatus 10, a new transfer unit 130 (e.g., a new gel layer 131d, a carrier film 131c, and/or upper and lower buffer medium layers), for example, control parameters thereof are rapidly set, so as to generate a suitable unipolar electric signal; in a daily multi-transfer process of a certain bio-transfer apparatus 10 for the same transfer unit 130, not only the control parameters of each transfer process can be optimized based on already obtained measurement information, transfer quality information, and the like, but also the transfer quality can be continuously improved. Moreover, the requirement on the professional ability of the operator can be greatly reduced by the self-learning function.

Specifically, the control parameter generating module 151 of the controller 150 may calculate or optimize the control parameters by using a kernel fitting algorithm, where the kernel fitting algorithm uses an artificial Neural Network, and the artificial Neural Network may be, but is not limited to, a Supervised Learning Network (super Learning Network), a Hybrid Learning Network (Hybrid Learning Network), a Reinforcement Learning Network (Reinforcement Learning Network), a Hopfield Network (Hopfield Network), a Boltzmann Machine (Boltzmann Machine), a Stochastic Neural Network (Stochastic Neural Network), and the like. The different control parameters can be optimized by setting different input variables to train the data of the neural network.

FIG. 9 is a schematic view showing the construction of a biological transfer apparatus according to a second embodiment of the present invention. As shown in fig. 9, the bio-transfer device 20 is also provided with one or more of a waveform generator 110, a voltage and current detection part 120, a transfer unit 130, a temperature sensor 140, a controller 150, a real-time clock 160, a human-computer interface 170 and a communication unit 180 similar to those of the bio-transfer device 10 shown in fig. 1, which are not described in detail herein, and the power module 190 provided in the bio-transfer device 20 has a relatively different implementation with respect to the bio-transfer device 10.

In one embodiment, as shown in FIG. 9, the power module 190 provides energy transfer via contactless electromagnetic coupling, which includes a primary unit 191 and a secondary unit 192, such that the bio-transfer device 20 can obtain electrical energy in a contactless manner. Therefore, the structure of the biological transfer device 20 in the embodiment shown in fig. 9 can be designed to be more compact, and it is easier to achieve overall water-proof, biological or chemical-proof (e.g., reducing metal contamination to biological agents and the like due to conductive contact), and the device is more convenient and flexible to use, and is very suitable for biological laboratories.

Fig. 10 is a schematic structural view of a biological transfer apparatus according to a third embodiment of the present invention. As shown in fig. 10, the bio-transfer device 20 is also provided with one or more of a waveform generator 110, a voltage and current detecting part 120, a transferring unit 130, a temperature sensor 140, a controller 150, a real-time clock 160, a human-machine interface 170, and a communication unit 180 similar to those of the bio-transfer device 10 shown in fig. 1, and detailed descriptions thereof will be omitted. The bio-transfer device 20 mainly has the following differences with respect to the bio-transfer device 10: (1) The bio-transfer device 20 is structurally divided into a control end 310 and a transfer end 320, the control end 310 and the transfer end 320 being separately provided; (2) The power module 190 has a relatively different implementation, and in particular, the energy and/or information transfer between the control terminal 310 and the transfer terminal 320 is achieved by contactless electromagnetic coupling.

As shown in fig. 10 in particular, the control end 310 is mainly used for implementing the function of a control unit, and is provided with a waveform generator 110, a voltage and current detection component 120, a controller 150, a real-time clock 160, a human-computer interaction interface 170 and a communication unit 180 similar to those in the bio-transfer device 10 shown in fig. 1, and an alternating current-direct current (AC-DC) conversion unit 193 for supplying power to the various components is further provided in the control end 310. An alternating current-direct current (AC-DC) conversion unit 193 is mainly used to convert AC power of a power grid into DC power of a corresponding level, and is part of a power module. Generated by the control terminal 310The single-polarity electrical signal (e.g., square wave voltage signal) applied to the transfer unit 130 is wirelessly transmitted to the transfer terminal 320, for example, can be simultaneously transmitted to a plurality (e.g., x being an integer greater than or equal to 2) of transfer terminals 320 ₁ 、320 ₂ ...320 _x . In addition to the transfer unit 130 and the secondary unit 192 of the power module, in one embodiment, a temperature sensor 140 is integrally disposed on each transfer end 320. For each transfer end 320, a primary unit 191 is provided on the control end 310, for example, x (x ≧ 2) primary units 191 are provided on one control end 310 ₁ 、191 ₂ ...191 _x . Thus, on one hand, the unipolar electric signal generated by the control terminal 310 can be transmitted to the plurality of transfer terminals 320 through contactless electromagnetic coupling, i.e., non-contact energy transmission is achieved, and on the other hand, the information generated by each transfer terminal 320, e.g., the temperature information collected by the temperature sensor 140, is transmitted to the control terminal 310 through contactless electromagnetic coupling, i.e., non-contact energy transmission is achieved.

Fig. 11 is a schematic structural diagram of a contactless electromagnetic coupling power supply module used in a biotransformation apparatus according to an embodiment of the present invention. The power module 190 in the bio-transfer device 30 includes a primary unit 191 and a secondary unit 192, a primary coil 1911 and a magnetic conductive shaft 1912 are disposed in the primary unit 191, a secondary coil 1921 is disposed in the secondary unit 192, and the primary unit 191 and the secondary unit 192 may be individually wrapped by, for example, plastic cases. When the magnetically permeable shaft 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 achieved therebetween based on the electromagnetic induction principle, so that the square wave voltage signal of the control terminal 310 can be transmitted to the transfer terminal 320; the temperature information of the transfer terminal 320 and the like may also be transmitted to the control terminal 310 in the form of a digital square wave.

In the bio-transfer device 30 in the embodiment shown in fig. 10, the structure of the transfer end 320 can be designed more compactly, and it is easier to achieve a waterproof design, biological or chemical contamination prevention (for example, to reduce metal contamination to biological reagents and the like due to conductive contact), and the use is more convenient and flexible, and is very suitable for being applied to a biological laboratory.

Fig. 12 is a schematic structural view of the first embodiment of the bio-transfer system formed based on the bio-transfer device of the embodiment shown in fig. 9. As shown in fig. 12, the bio-transfer system includes a plurality of bio-transfer devices 20, and the control units thereof are coupled to the cloud server 90, so that information or data transmission can be realized between the cloud server 90 and any one of the bio-transfer devices 20, for example, measurement information and/or transfer quality information obtained in the bio-transfer device 20 can be uploaded to the cloud server 90, and the cloud server 90 can also transmit information (e.g., control parameters) to the corresponding bio-transfer device 20.

In one embodiment, n (n ≧ 2) bioprinting devices 20 ₁ 、20 ₂ ...20 _n Can be communicatively connected to one portable intelligent terminal 175 at the same time, for example, to the portable intelligent terminal 175 through the communication unit 180 of the bio-transfer device 20, the portable intelligent terminal 175 may be specifically a tablet computer (IPAD), a smart phone, etc., and the portable intelligent terminal 175 may implement all or part of the functions of the human-machine interface 170 of the bio-transfer device 20, for example, by installing a corresponding APP application thereon, to interact with each bio-transfer device 20 (particularly, the controller 150).

In one embodiment, the cloud server 90 can communicate with m (m ≧ 2) portable intelligent terminals 175 ₁ ...175 _m Is connected in communication, thereby realizing communication with m portable intelligent terminals 175 ₁ ...175 _m Any of the attached bio-transfer devices 20 is coupled; in yet another alternative embodiment, the cloud server 90 may also be directly communicatively connected to the transfer device 20.

Fig. 13 is a schematic view showing the configuration of a second embodiment of a bio-transfer system formed based on the bio-transfer device of the embodiment shown in fig. 10. As shown in FIG. 13, the cloud server 90 of the bio-transfer system and m (m ≧ 2) portable intelligent terminals 175 ₁ ...175 _m Each intelligent terminal 175 is connected with n (n is more than or equal to 2) biological transfer printing devices 30 in a communication way ₁ 、30 ₂ ...30 _n And (4) communication connection.

Fig. 14 is a schematic block diagram illustrating a cloud server of the bio-transfer 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 may store data sent by each of the plurality of bio-transfer devices, and the history database 910 may store at least measurement information or/and transfer quality information records obtained by each of the plurality of bio-transfer devices 20 or 30 at each transfer process as history data information (for example, data stored by the storage module 153 of each bio-transfer device may be sent to the history database 910), so that, as the transfer process is continuously performed, a huge amount of history data information may be formed, and more history data information may provide more and more help for accuracy and effectiveness of cloud computing; the cloud computing module 920 may perform cloud computing or big data analysis on the historical data, and specifically, generate a control parameter corresponding to a current transfer process of one of the plurality of bio-transfer devices (10, 20, or 30) based on historical data information, so as to facilitate quick determination of a control parameter corresponding to the current transfer process using the historical data information, thereby greatly improving user experience.

It should be understood that, during the computing and analyzing process, the cloud computing module 920 may apply various cloud technologies to enhance robustness of the intelligent control, and the history database 910 may selectively store the measurement information and/or the control parameters corresponding to the transfer process with better transfer quality information, so that during the cloud computing process, the efficiency of obtaining the optimal control parameters through self-learning may be accelerated, and the cloud computing workload may be reduced. In an embodiment, the cloud computing module 920 may also replace, in whole or in part, the functions of the control parameter generation module 151 in the controller 150.

As shown in fig. 9, the control parameters obtained by the cloud computing module 920 may be transmitted to the control unit of the corresponding bio-transfer device 10, 20, or 30 through the transmission module 193. The bio-transfer device 10, 20, or 30 may generate or update the corresponding unipolar electrical signal based on the control parameter.

It should be understood that, due to the introduction of the cloud server 90, the historical data information of many biological transfer devices is shared, and the control parameters are obtained through the calculation of the cloud computing module 920 on the historical data information, which is equivalent to the fact that the experience of each user in setting the control parameters is shared and mined, so that the requirements of the professional skills and experience of the user or experimenter are greatly reduced.

It will be understood that when an element is referred to as being "connected," "coupled," or "coupled" to another element, it can be directly connected, coupled, or coupled to the other element or intervening elements may be present.

The above examples mainly describe the bio-transfer device and the control method thereof, and the bio-transfer system formed based on a plurality of bio-transfer devices according to the present invention. Although only a few embodiments of the present invention have been described, those skilled in the art will appreciate that the present invention may be embodied in many other forms without departing from the spirit or scope thereof. Accordingly, the present examples and embodiments are to be considered as illustrative and not restrictive, and various modifications and substitutions may be made therein without departing from the spirit and scope of the present invention as defined by the appended claims.

Claims (24)

1. A bio-transfer device comprising a control unit and one or more transfer units, wherein the control unit is configured to: and applying a unipolar electric signal which periodically fluctuates according to a predetermined amplitude to the first and second plate electrode layers of one or more transfer units, wherein the 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 2.

2. The bio-transfer device according to claim 1, wherein the predetermined amplitude is constant or varies during the periodic fluctuation of the unipolar electric signal.

3. The bio-transfer device according to claim 1, wherein the undulating unipolar electric signal includes 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. The bio-transfer device according to claim 1, wherein the undulating unipolar electric signal comprises a square wave voltage signal with an adjustable duty ratio, or at least one of an intermittent sine wave signal, a triangular wave signal, a sawtooth wave signal, and a staircase signal.

5. The biological transfer device of claim 3 or 4, wherein the frequency of the unipolar electrical signal is greater than or equal to 1Hz and less than or equal to 100Hz, or greater than or equal to 5Hz and less than or equal to 20Hz.

6. The biological transfer device of claim 3 or 4, wherein the unipolar electrical signal has a duty cycle of 1% to 99%, or 30% to 60%.

7. The bio transfer device according to claim 3 or 4, wherein the unipolar electric signal is a voltage signal having a voltage peak value greater than or equal to 1V and less than or equal to 30V, and a peak current generated on at least one of the transfer units corresponding to the voltage peak value is greater than or equal to 0.1A and less than or equal to 10A.

8. The biological transfer device of claim 1, wherein the transfer unit comprises:

the first flat electrode layer and the second flat electrode layer are arranged in parallel; and

the first buffer dielectric layer, the gel layer containing biological macromolecules, the bearing film and the second buffer dielectric layer are positioned between the first flat electrode layer and the second flat electrode layer;

wherein the biological macromolecules in the gel layer are electrophoresed to the carrier film under the action of the applied unipolar electric signal.

9. The biological transfer apparatus of claim 8, wherein a ratio of the electrical resistance of the first buffer dielectric layer, the second buffer dielectric layer, and the carrier film to the electrical resistance of the gel layer is less than or equal to 3.

10. The bio-transfer device according to claim 9, further comprising a temperature sensor for measuring temperature information of the first flat electrode layer and/or the second flat electrode layer in real time during the transfer process;

wherein the temperature information is fed back to the control unit, and the control unit is further configured to: dynamically adjusting a duty cycle of the unipolar signal during a transfer process based at least on the temperature information.

11. The biological transfer device of claim 10, wherein the control unit is further configured to: and adjusting at least the duty ratio to enable the temperature of the transfer unit to be lower than 60-70 ℃ in the transfer process.

12. The bio-transfer 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 bioconversion device of claim 1, wherein the bioconversion device is a western blot device or a southern blot device.

14. The biological transfer device of claim 1, wherein said control unit comprises:

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

a unipolar electric signal generation module for generating the unipolar electric signal according to the adjusted control parameter;

wherein the adjusted control parameter comprises at least one of a waveform, a frequency, a voltage value, a current value, and a duty cycle of the unipolar electric signal;

wherein the measurement information is temperature information of the first flat electrode layer and/or the second flat electrode layer measured in real time during the transfer process, or timing information including at least one of a waveform, a frequency, a voltage value, a current value, and a duty ratio of the unipolar electric signal, and temperature information of the first flat electrode layer and/or the second flat electrode layer, which is recorded during the transfer process.

15. The biological transfer device of claim 14, wherein said control unit further includes:

the input module is used for receiving the input transfer printing quality information; and

and the storage module is used for recording the measurement information or/and the transfer quality information of each transfer process.

16. A bio-transfer system, comprising:

a plurality of the biological transfer devices of any one of claims 1-15;

a cloud server coupled to control units of a plurality of the bio-transfer devices;

wherein the cloud server is configured to include:

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

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

a sending module for sending the control parameters to the control unit of the corresponding biological transfer printing device;

wherein the control parameter comprises at least one of a waveform, a frequency, a voltage value, a current value, and a duty cycle of the unipolar electric signal; the measurement information is temperature information of the first flat electrode layer and/or the second flat electrode layer measured in real time in a transfer process, or timing information including at least one of a waveform, a frequency, a voltage value, a current value, and a duty ratio of the unipolar electric signal, and temperature information of the first flat electrode layer and/or the second flat electrode layer, which is recorded in the transfer process.

17. A control method for a bio-transfer apparatus including one or more transfer units including first and second plate electrode layers, and a gel layer and a carrier film between the first and second plate electrode layers; the transfer unit is characterized in that a unipolar electric signal which periodically fluctuates according to a predetermined amplitude is uniformly applied to the first flat electrode layer and the second flat electrode layer of one or more transfer units in a transfer process, wherein the 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 2.

18. The control method of 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 of claim 17, wherein the fluctuating unipolar electric signal comprises a duty-cycle adjustable square wave voltage signal, 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, characterized in that the frequency of the fluctuations of the unipolar electric signals is greater than or equal to 1Hz and less than or equal to 100Hz, or greater than or equal to 5Hz and less than or equal to 20Hz.

21. A control method according to claim 18 or 19, wherein the duty cycle of the unipolar electric signal is between 1% and 99%, or between 30% and 60%.

22. The control method according to claim 18 or 19, wherein the unipolar electric signal is a voltage signal having a voltage peak greater than or equal to 1V and less than or equal to 30V, a peak current generated on at least one of the transfer units corresponding to the voltage peak being greater than or equal to 0.1A and less than or equal to 10A.

23. The control method according to claim 21, characterized by further comprising:

generating adjusted control parameters based on the measurement information or/and the transfer quality information, the adjusted control parameters including at least one of a waveform, a frequency, a voltage value, a current value, and a duty ratio of the unipolar electric signal; and

updating the unipolar electric signal according to the adjusted control parameter, and applying the updated unipolar electric signal to the transfer unit.

24. The control method of claim 23, prior to the generating step, further comprising:

measuring temperature information of the first flat electrode layer and/or the second flat electrode layer in real time in a transfer process as the measurement information; or

Recording timing information including at least one of a waveform, a frequency, a voltage value, a current value and a duty ratio of the unipolar electric signal, and temperature information as the measurement information during a transfer process, wherein the temperature information is obtained by measuring a temperature of the first flat electrode layer and/or the second flat electrode layer in real time during the transfer process.

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