CN218823932U - Alzheimer's early sieve device based on ultrasonic accelerated reaction and aggregation enhancement - Google Patents
Alzheimer's early sieve device based on ultrasonic accelerated reaction and aggregation enhancement Download PDFInfo
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- CN218823932U CN218823932U CN202222859187.3U CN202222859187U CN218823932U CN 218823932 U CN218823932 U CN 218823932U CN 202222859187 U CN202222859187 U CN 202222859187U CN 218823932 U CN218823932 U CN 218823932U
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Abstract
The utility model discloses an Alzheimer's early sieve device based on ultrasonic accelerated reaction and aggregation enhancement, which comprises a packaging glass sheet, a micro-fluidic chip, a steel sheet and an ultrasonic piezoelectric ceramic sheet which are arranged from top to bottom in sequence; the detection device comprises a package glass sheet, a steel sheet, a micro-fluidic chip and a micro-fluidic chip, wherein a Kapton adhesive tape layer is arranged between the package glass sheet and the steel sheet, the micro-fluidic chip is arranged in the Kapton adhesive tape layer, and the micro-fluidic chip comprises a circular micro-cavity for containing detection liquid; the device also comprises a waveform generator and a power amplifier, wherein the waveform generator is used for generating sine waves and is connected with the power amplifier through a lead, and the power amplifier is used for generating amplified voltage and applying the amplified voltage to the ultrasonic piezoelectric ceramic chip to generate ultrasonic waves. The utility model discloses introduce supersound drive technology and realize the acceleration reaction and the gathering reinforcing to catalysis hairpin self-assembly system to effectively improve detectivity and shorten check-out time.
Description
Technical Field
The utility model relates to an analysis and detection technical field, in particular to early sieve device of alzheimer's disease based on supersound accelerating reaction and gathering reinforcing.
Background
Alzheimer's Disease (AD) is a progressive neurodegenerative disease, and currently about 4680 million people worldwide suffer from AD. With the aging of the world population further aggravated, the number of AD patients has increased dramatically, and AD has become the third leading cause of disability and death in the elderly, second only to cardiovascular and cerebrovascular diseases and malignant tumors.
However, AD is a complex disease involving multiple factors. Due to the complexity of the human brain, the lack of a reasonable animal model and research tools for its study, the detailed pathogenesis of AD remains unclear to date. At present, alzheimer's disease still is a global medical problem, the detection method is very limited, and the early identification and screening of the Alzheimer's disease are important.
The existing early AD screening means mainly comprises a fluorescence detection method, and a micro-fluidic chip is used for carrying out fluorescence spectrum detection on a catalysis hairpin self-assembly (CHA) system, so that the early Alzheimer screening is realized. However, how to improve the sensitivity of the detection device and shorten the detection time is an urgent technical problem to be solved in the field.
SUMMERY OF THE UTILITY MODEL
To the problem, an object of the utility model is to provide an early sieve device of alzheimer's disease based on supersound is accelerated reaction and gathering reinforcing realizes the accelerated reaction and the gathering reinforcing to catalysis hairpin self-assembly (CHA) system through introducing supersound drive technology to effectively improve detectivity and shorten check-out time, realize the quick examination of early high sensitivity of alzheimer's disease.
In order to solve the above technical problem, the embodiment of the utility model provides a following scheme:
an Alzheimer's early sieve device based on ultrasonic accelerated reaction and aggregation enhancement comprises a packaging glass sheet, a micro-fluidic chip, a steel sheet and an ultrasonic piezoelectric ceramic sheet which are sequentially arranged from top to bottom;
the detection device comprises a packaging glass sheet, a steel sheet, a micro-fluidic chip and a micro-fluidic chip, wherein a Kapton adhesive tape layer is arranged between the packaging glass sheet and the steel sheet, the micro-fluidic chip is arranged in the Kapton adhesive tape layer, and comprises a circular micro-cavity for containing detection liquid;
the device also comprises a waveform generator and a power amplifier, wherein the waveform generator is used for generating sine waves and is connected with the power amplifier through a lead, and the power amplifier is used for generating amplified voltage and applying the amplified voltage to the ultrasonic piezoelectric ceramic chip to generate ultrasonic waves.
Preferably, the device further comprises a temperature sensing controller, wherein the temperature sensing controller is used for monitoring and regulating and controlling the temperature in the circular micro cavity in real time.
Preferably, the device further comprises an oscilloscope, wherein the oscilloscope is connected with the waveform generator and is used for monitoring the waveform signal generated by the waveform generator.
Preferably, the diameter of the circular microcavity is 5mm.
Preferably, the frequency of the sine wave generated by the waveform generator is between 500kHz and 1000 kHz.
Preferably, the power amplifier applies a voltage of between 0V and 10V.
The embodiment of the utility model provides a beneficial effect that technical scheme brought includes at least:
the utility model discloses realize the acceleration reaction and the gathering reinforcing to catalysis hairpin self-assembly (CHA) system based on ultrasonic drive technique, through producing the ultrasonic wave and acting on the self-amplification rate who accelerates the solute in the reaction system, realized detecting the trace enrichment of the sample that awaits measuring, have that detectivity is high, detect that the limit is low, required check-out time is short waiting advantage, have important value in the aspect of realizing the quick early examination of Alzheimer.
Drawings
In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings used in the description of the embodiments will be briefly described below, and it is obvious that the drawings in the description are only some embodiments of the present invention, and it is obvious for those skilled in the art that other drawings can be obtained without creative efforts.
Fig. 1 is a schematic structural diagram of an alzheimer's early-stage sieve device based on ultrasonic accelerated reaction and aggregation enhancement provided by an embodiment of the present invention;
FIG. 2 is a schematic diagram of a circular microcavity in an embodiment of the present invention;
FIGS. 3a and 3b are schematic diagrams illustrating the comparison of fluorescence intensity of the probe-modified particles in the dispersed state and the central aggregation state according to the embodiment of the present invention.
Description of reference numerals: 1-encapsulating the glass sheet; 2-a microfluidic chip; 3-steel sheet; 4-ultrasonic piezoelectric ceramic plate; 5-Kapton adhesive tape layer; 6-a waveform generator; 7-a power amplifier; 8-oscilloscope.
As shown in the drawings, in order to clearly realize the structures of the embodiments of the present invention, specific structures and devices are labeled in the drawings, but this is only for illustration purpose, and it is not intended to limit the present invention to the specific structures, devices and environments, and those skilled in the art can adjust or modify these devices and environments according to specific needs, and the adjustment or modification is still included in the protection scope of the present invention.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention clearer, embodiments of the present invention will be described in further detail below with reference to the accompanying drawings.
The embodiment of the utility model provides an advanced screening device of Alzheimer's disease based on ultrasonic accelerated reaction and gathering reinforcing, as shown in figure 1, the device includes encapsulation glass piece 1, micro-fluidic chip 2, steel sheet 3 and supersound piezoceramics piece 4 that set gradually from top to bottom;
wherein, a Kapton adhesive tape layer 5 is arranged between the encapsulation glass sheet 1 and the steel sheet 3, the microfluidic chip 2 is arranged in the Kapton adhesive tape layer 5, and the microfluidic chip 2 comprises a round microcavity for containing detection liquid, as shown in fig. 2;
the device also comprises a waveform generator 6 and a power amplifier 7, wherein the waveform generator 6 is used for generating sine waves and is connected with the power amplifier 7 through a lead, and the power amplifier 7 is used for generating amplified voltage and applying the amplified voltage to the ultrasonic piezoelectric ceramic piece 4 to generate ultrasonic waves.
Further, the device also comprises a temperature sensing controller (not shown in the figure), and the temperature sensing controller is used for monitoring and regulating the temperature in the circular micro cavity in real time so as to keep the temperature at a proper temperature. The position of the temperature sensing controller can be set according to specific situations and actual requirements, and is not limited herein.
Further, the device also comprises an oscilloscope 8, wherein the oscilloscope 8 is connected with the waveform generator 6 and is used for monitoring the waveform signal generated by the waveform generator 6.
The embodiment of the utility model provides an in, encapsulation glass piece 1 is used for sealed micro-fluidic chip 2 and plays the effect at circular microcavity internal reflection ultrasonic wave, the diameter of circular microcavity is about 5mm, and steel sheet 3 is used for transmitting the ultrasonic wave and plays the effect of filtering, and supersound piezoceramics piece 4 produces sinusoidal ultrasonic wave through piezoelectric conversion and acts on inside circular microcavity. The temperature sensing controller is used for monitoring the temperature in the circular micro cavity in real time and regulating and controlling the temperature to keep the temperature at a proper temperature.
The ultrasonic piezoelectric ceramic plate 4 is connected with a waveform generator 6 capable of generating an arbitrary waveform and a power amplifier 7. Continuous sine waves are generated by the waveform generator 6 and are connected with the power amplifier 7 through a lead, and the generated amplified voltage is applied to the ultrasonic piezoelectric ceramic piece 4 to generate ultrasonic waves. The ultrasonic wave passes through the steel sheet 3 to reach the microfluidic chip 2 containing the circular micro-cavity, and all experiments are completed in the circular micro-cavity of the microfluidic chip 2.
In this embodiment, the frequency of the continuous sinusoidal waveform generated by the waveform generator 6 is in the range of 500kHz to 1000kHz, the amplitude of the voltage applied by the power amplifier 7 is in the range of 0V to 10V, and the waveform signal is monitored by the oscilloscope 8. The magnitude of the applied ultrasonic wave can be controlled by controlling the voltage applied to the ultrasonic piezoelectric ceramic sheet 4. Meanwhile, the temperature in the circular micro cavity is monitored and controlled in real time through a temperature sensing controller, so that the temperature is always kept at room temperature.
Sine waves emitted by the waveform generator 6 are connected to the ultrasonic piezoelectric ceramic plate 4 after passing through the power amplifier 7 to generate ultrasonic waves, and the ultrasonic waves are transmitted upwards by penetrating through the steel sheet 3 to generate reflection on the inner wall of the circular micro-cavity. The reflection here includes not only the reflection of the upper encapsulation glass sheet 1 but also the reflection of the left and right inner walls of the circular microcavity. The movement of the focus point in the experiment is the result of the reflection of the ultrasonic wave by the inner wall, and the superposition of the two ultrasonic waves reflected by the inner wall with the same frequency and amplitude but opposite propagation directions generates a plane standing wave, thereby moving the nano particles.
In the device, an appropriate amount of sample liquid with a fixed concentration is input to the center of the circular micro-cavity by using an input tube, and the frequency of ultrasonic waves is adjusted by controlling the current frequency of a signal generator 6 and the voltage generated by a power amplifier 7. In a proper ultrasonic range, the surface acoustic wave provides a stable driving force for the surface of the sample liquid in the circular micro-cavity, so that the solute in the liquid drop to be detected is accelerated to gather.
The utility model discloses creatively build the amplification system, the ultrasonic agitation gathers and the integration of three techniques of fluorescence detection is in the same place, according to the fluorescence intensity that each sample after the amplification that observes in fluorescence microscope behind the ultrasonic aggregation corresponds, draws fluorescence gradient image and curve to confirm sample concentration fast accurately. Therefore, the device can be applied to occasions such as the early screening of the Alzheimer of key crowds, provides a novel detection device which has higher sensitivity, more accurate detection result and shorter detection time compared with the prior detection technology, and obtains an accurate experimental result.
Compare in traditional amplification technique, the utility model discloses can adapt to more complicated external environment, need not harsh amplification reaction condition, shorten reaction time, improve the sensitivity and the stability of testing result.
Based on the device carries out the concrete implementation process that the Alzheimer's marker detected as follows:
first, the PS microspheres used for capture probes were modified:
based on the base complementary pairing method, equal amounts of 1-2. Mu.L of target probe and H1 were mixed for about 15 minutes to pair as a new probe. In the next stage, 1 μ L of H2 probe is added to replace the target probe and act on the new probe, the fluorescently labeled H2 probe is anchored on the functionalized and modified 1 μ L of PS microspheres, the reaction is carried out for 30 minutes, during which the target probe is released, and the released target can induce the next cycle. In the reaction process, after H1 and H2 are combined into a new probe, the fluorescence signal at the tail end of H2 is separated from the quenching group, the fluorescence signal is released after a single fluorescence group leaves the quenching group, and the fluorescence signal is anchored on the functionalized modified PS microsphere, so that the subsequent operation is facilitated. In the experiment, the fluorescence signal of the PS microspheres which are gathered and provided with the fluorescence-labeled H2 probe is amplified exponentially, so that the fluorescence imaging and detection of the nucleic acid detection result can be conveniently and rapidly carried out.
Sample probe sequences required in this experiment:
H1：5’-C TTC AGT TAT CAC AGT ACT GTA GT GGT GTA TAC AGT ACT GTG-3’；
H2：5’-ACT GTA-BHQ1-TAC TAC AGT ACT GTG GAT GT GGT GTA-FAM-3’；
target sequence(miRNA-101)：5′-UAC AGU ACU GUG AUA ACU GAA G-3′。
and then, starting the device to perform ultrasonic stirring to accelerate the reaction for 5-15 minutes, and adjusting the frequency of the ultrasonic wave to reflect the ultrasonic wave in the device so as to realize the aggregation of the functionalized modified PS microspheres. In the ultrasonic container system, sound waves pass through the steel sheet through the ultrasonic piezoelectric ceramic sheet and are transmitted upwards, and reflection is generated on the inner wall of the circular micro-cavity, so that plane standing waves are generated by overlapping the sound waves in opposite directions, particles in a sample are pushed to the center of the cavity, and the sample is subjected to center aggregation. The speed of aggregation of the whole functionalized and modified microsphere can be realized by adjusting the applied voltage, and the specific position of the motor aggregation can be controlled by changing the position of an ultrasonic node or an antinode by changing the applied frequency.
After the sample is stirred and aggregated in the device without contact by using ultrasonic waves, fluorescence detection is carried out on an aggregation point by using a fluorescence microscope, and the concentration of the Alzheimer's disease marker to be detected is obtained by measuring the fluorescence intensity of the aggregation point. The results of comparing the fluorescence intensity of the probe-modified particles in the dispersed state and the central aggregation state are shown in fig. 3a and 3 b. Experiments prove that the device is used for AD early screening detection, can exponentially multiply the concentration and the fluorescence intensity of a strong target, shortens the amplification time, and greatly improves the detection efficiency.
It should be noted that, in this document, the terms "comprises," "comprising," or any other variation thereof, are intended to cover a non-exclusive inclusion, such that a process, method, article, or terminal apparatus that comprises a list of elements does not include only those elements but may include other elements not expressly listed or inherent to such process, method, article, or terminal apparatus. Without further limitation, an element defined by the phrases "comprising one of \ 8230; \8230;" does not exclude the presence of additional like elements in a process, method, article, or terminal device that comprises the element.
References in the specification to "one embodiment," "an example embodiment," "some embodiments," etc., indicate that the embodiment described may include a particular feature, structure, or characteristic, but every embodiment may not necessarily include the particular feature, structure, or characteristic. Further, when a particular feature, structure, or characteristic is described in connection with an embodiment, it is submitted that it is within the knowledge of one skilled in the relevant art to effect such feature, structure, or characteristic in connection with other embodiments whether or not explicitly described.
It is understood that the meaning of "on 8230; \8230on," \8230, above "and" on 82308230; \823030, above "in the present disclosure should be interpreted in the broadest manner such that" on 8230; \8230above "means not only" directly on "something" but also on "something with the meaning of intervening features or layers therebetween, and" on 8230; \8230on "or" on 8230, above "not only means" on "or" above "something, but also may include the meaning thereof" on "or" above "something with no intervening features or layers therebetween.
Furthermore, spatially relative terms such as "below 8230; below," "lower," "above 8230; above," "upper," and the like may be used herein for ease of description to describe one element or feature's relationship to another element or feature or features, as illustrated in the figures. Spatially relative terms are intended to encompass different orientations in use or operation of the device in addition to the orientation depicted in the figures. The device may be otherwise oriented and the spatially relative descriptors used herein interpreted accordingly.
As used herein, the term "about" indicates a value of a given amount that can vary based on the particular technology node associated with the subject device. The term "about" may indicate a given amount of a value that varies, for example, within 5% -15% of the value (e.g., ± 5%, ± 10% or ± 15% of the value), based on the particular technology node.
The present invention covers any alternatives, modifications, equivalents, and alternatives falling within the spirit and scope of the invention. In the following description of the preferred embodiments of the present invention, specific details are set forth in order to provide a thorough understanding of the present invention, and it will be apparent to those skilled in the art that the present invention may be practiced without these specific details. In other instances, well-known methods, procedures, flows, components, circuits, and the like have not been described in detail as not to unnecessarily obscure aspects of the present invention.
The above description is only for the preferred embodiment of the present invention, and is not intended to limit the present invention, and any modifications, equivalent replacements, improvements, etc. made within the spirit and principle of the present invention should be included within the protection scope of the present invention.
Claims (6)
1. An Alzheimer's early sieve device based on ultrasonic accelerated reaction and aggregation enhancement is characterized by comprising a packaging glass sheet, a micro-fluidic chip, a steel sheet and an ultrasonic piezoelectric ceramic sheet which are sequentially arranged from top to bottom;
the detection device comprises a packaging glass sheet, a steel sheet, a micro-fluidic chip and a micro-fluidic chip, wherein a Kapton adhesive tape layer is arranged between the packaging glass sheet and the steel sheet, the micro-fluidic chip is arranged in the Kapton adhesive tape layer, and comprises a circular micro-cavity for containing detection liquid;
the device also comprises a waveform generator and a power amplifier, wherein the waveform generator is used for generating sine waves and is connected with the power amplifier through a lead, and the power amplifier is used for generating amplified voltage and applying the amplified voltage to the ultrasonic piezoelectric ceramic chip to generate ultrasonic waves.
2. The alzheimer's early sieve device based on ultrasonically accelerated reaction and aggregation enhancement as claimed in claim 1, further comprising a temperature sensing controller for monitoring and controlling the temperature inside the circular micro-cavity in real time.
3. The Alzheimer's promethazine device based on the ultrasonic accelerated reaction and aggregation enhancement according to claim 1, further comprising an oscilloscope connected with the waveform generator for monitoring the waveform signal generated by the waveform generator.
4. The alzheimer's promyelocytic ultrasound screen apparatus based on ultrasonically accelerated reaction and aggregation enhancement of claim 1, wherein the circular microcavity has a diameter of 5mm.
5. The alzheimer's early sieve device based on ultrasonic accelerated reaction and aggregation enhancement of claim 1, wherein the frequency of the sine wave generated by the waveform generator is between 500kHz and 1000 kHz.
6. The alzheimer's promethazine device based on ultrasonically accelerated reaction and aggregation enhancement according to claim 1, wherein the amplitude of the voltage applied by the power amplifier is between 0V and 10V.
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