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WO2023004795A1 - Appareil et procédé de pipetage ultrasonore - Google Patents

Appareil et procédé de pipetage ultrasonore Download PDF

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Publication number
WO2023004795A1
WO2023004795A1 PCT/CN2021/109810 CN2021109810W WO2023004795A1 WO 2023004795 A1 WO2023004795 A1 WO 2023004795A1 CN 2021109810 W CN2021109810 W CN 2021109810W WO 2023004795 A1 WO2023004795 A1 WO 2023004795A1
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WO
WIPO (PCT)
Prior art keywords
ultrasonic
liquid
coupling
probe
pipetting
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Ceased
Application number
PCT/CN2021/109810
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English (en)
Chinese (zh)
Inventor
张志强
潘东文
邱维宝
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Shenzhen Insightsonics Co Ltd
Shenzhen Institute of Advanced Technology of CAS
Original Assignee
Shenzhen Insightsonics Co Ltd
Shenzhen Institute of Advanced Technology of CAS
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Shenzhen Insightsonics Co Ltd, Shenzhen Institute of Advanced Technology of CAS filed Critical Shenzhen Insightsonics Co Ltd
Priority to PCT/CN2021/109810 priority Critical patent/WO2023004795A1/fr
Publication of WO2023004795A1 publication Critical patent/WO2023004795A1/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

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    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
    • B01L3/02—Burettes; Pipettes

Definitions

  • the invention relates to the technical field of synthetic biology experiments, in particular to an ultrasonic pipetting device and method.
  • Pipettes of varying accuracy and precision can be achieved through a variety of designs, ranging from simple pipettes made from a single piece of glass to complex adjustable or electronically controlled pipettes.
  • the accuracy of its measurement varies greatly from species to species.
  • the pipette gun is a contact pipetting method, during the pipetting process, the sample adheres to the tip of the pipette, resulting in inaccurate amount of transferred liquid and prone to false negative results.
  • the present invention proposes an ultrasonic pipetting device and method that can use multiple probes to pipette simultaneously without interfering with each other.
  • the ultrasonic excitation component is connected to a control circuit, and the ultrasonic excitation component includes two or more ultrasonic excitation units, and each of the ultrasonic excitation units is connected to a corresponding probe for exciting each of the probes, and the probes are used for emit ultrasound. Due to the surface adhesion of the liquid, when the coupling liquid touches the bottom of the liquid-carrying device, it will be connected to form an ultrasonic channel composed of the coupling liquid.
  • the ultrasonic excitation unit includes a drive circuit and an excitation circuit, the drive circuit is connected to the control circuit, the excitation circuit is connected to the probe, the control circuit controls the drive circuit to generate ultrasonic waves, and the excitation circuit controls the
  • the probes are used for excitation; the different ultrasonic excitation units are independent of each other, and are used to simultaneously excite multiple probes to emit ultrasonic waves at the same or different frequencies.
  • the probe is a focused ultrasonic transducer, and the ultrasonic waves emitted by the probe are focused ultrasonic waves.
  • the multi-probe assembly includes a plurality of motors, each of the motors is connected to the corresponding probe, and the motor drives the probe to change the distance between the probe and the liquid-carrying device .
  • the multi-probe assembly further includes a housing tank connected to the coupling liquid circulation system; the housing tank surrounds the coupling cavity, and the coupling liquid overflowing the coupling nozzle flows into the housing groove.
  • the coupling liquid in the coupling liquid circulation system continuously enters the multi-probe assembly, and the coupling liquid overflowing from the coupling nozzle flows back to the coupling liquid circulation system, which can realize the recycling of the coupling liquid, and can replace the coupling liquid in the multi-probe assembly in real time, so as to A better effect of cooling the probe is achieved.
  • the ultrasonic transmitting module also includes a signal detection component connected to the control circuit for receiving the detection signal of the probe; the signal detection component includes two or more signal detection channels, each of the The signal detection channel is connected to the corresponding probe.
  • the processing circuits and acquisition circuits of different signal detection channels are independent of each other, so the parameters of each probe can be obtained independently without affecting each other, and can support simultaneous detection of multiple channels.
  • the signal detection component can obtain data such as the liquid level, volume and solution type of the liquid carrier device through the detection signal, and can automatically obtain the physical parameters of the liquid.
  • the ultrasonic excitation component also includes a power detection unit, the power detection unit includes a signal coupling module and a signal acquisition module, and the signal acquisition module measures the attenuated signal of the ultrasonic excitation component coupled by the signal coupling module to obtain an ultrasonic Excitation signal amplitude.
  • the present invention also provides an ultrasonic pipetting method, which uses the above-mentioned ultrasonic pipetting device to perform ultrasonic pipetting, and the ultrasonic pipetting method includes:
  • the ultrasonic excitation unit excites different probes to emit ultrasonic waves at the same or different frequencies.
  • the ultrasonic pipetting device and method of the present invention have the following beneficial effects: each probe is connected to a different ultrasonic excitation unit, each ultrasonic excitation unit can work independently, and supports the excitation and setting of multiple ultrasonic waveform parameters, To meet the needs of various incentive parameters. Therefore, the ultrasonic liquid pipetting device of the present invention can coordinate the work of multiple probes with the same acoustic parameters, and can also support multiple probes with different acoustic parameters to work together for ultrasonic liquid pipetting, and can flexibly control the size of each pipetting liquid, providing high-quality Precision control and pipetting efficiency provide a better solution.
  • each probe is equipped with a motor to drive the probe to move up and down to change the distance between the probe and the liquid surface of the liquid carrier device.
  • Multiple probes can work at the same time and each probe can focus independently. Multiple probes can be flexibly matched.
  • the ultrasonic pipetting method carried out by the ultrasonic pipetting device of the present invention can coordinate the work of a plurality of probes with the same acoustic parameters to improve the speed and efficiency of ultrasonic pipetting; it can also support the simultaneous work of probes with different acoustic parameters to realize the pipetting function, to flexibly control the size of each pipette.
  • Fig. 1 is the schematic structural view of the multi-probe assembly of the ultrasonic pipetting device of the present invention
  • Fig. 2 is a schematic structural view of the coupling liquid circulation system of the ultrasonic pipetting device of the present invention
  • Fig. 3 is a structural schematic diagram of the ultrasonic excitation assembly of the ultrasonic pipetting device of the present invention.
  • Fig. 4 is a schematic structural diagram of a signal detection component of the ultrasonic pipetting device of the present invention.
  • the multi-probe assembly is an integrated structure, including a bracket 11, a motor 12, a coupling chamber 13, a housing groove 14, a coupling nozzle 15, a probe 16, and a sealing tube 17, wherein the number of probes 16 is two and above.
  • the coupling cavity 13 surrounds the probe 16
  • the accommodating groove 14 surrounds the coupling cavity 15 .
  • This embodiment provides a specific structure of a dual-probe assembly: two probes 16 are arranged on the same bracket 11, and a coupling cavity 13 is provided on the outside of each probe 16, and the coupling cavity 13 is filled with coupling liquid, and the coupling cavity 13 surrounds
  • the probe 16 and the accommodating groove 14 surround the two coupling cavities 13, both the accommodating groove 14 and the coupling cavities 13 are fixed on the support 11, and the two coupling cavities 13 are separated by a metal plate.
  • the coupling liquid circulation system includes a sealed tank 31 , a water pump 32 , a temperature control system 33 , a thermometer 34 , a filter 35 , a vacuum pump 36 and a muffler 37 .
  • the inlet port of the sealed tank 31 is connected with a filter 35
  • the outlet port is connected with a water pump 32
  • the sealed tank 31 is also connected with a vacuum pump 36
  • a muffler 37 is set to connect the vacuum pump 36 for reducing the noise generated by the vacuum pump 36.
  • the water pump 32, temperature Control system 33 and thermometer 34 are connected in sequence.
  • the coupling nozzle 15 is a hollow structure, preferably a hollow conical structure.
  • the coupling nozzle 15 is arranged between the probe 16 and the liquid-carrying device 2.
  • the bottom of the coupling nozzle 15 is open to fit into the coupling chamber 13, and an opening is left on the top.
  • the opening surrounds the probe 16 and its caliber is larger than the diameter of the probe 16.
  • the coupling nozzle 15 and the probe 16 to form a gap.
  • the coupling liquid may be water.
  • the coupling liquid circulation system makes the coupling liquid continuously enter the coupling chamber 13. After the coupling liquid fills the coupling chamber 13, it flows out from the small opening on the top of the coupling nozzle 15.
  • the probe 16 is connected with the motor 12, and the bottom of the coupling cavity 13 is provided with a through hole for the probe 16 to pass through.
  • One end of the sealing tube 17 seals the through hole at the bottom of the coupling cavity 13, and the other end is tightly combined with the side wall of the probe 16.
  • the motor 12 drives the probe 16 to move up and down to change the distance between the liquid surface of the liquid carrying device 2; the sealing tube 17 can be folded or stretched to follow the probe 16 to move up and down when the motor 12 drives the probe 16 to move up and down, so that the coupling cavity 13
  • the bottom remains sealed to prevent the coupling liquid from entering the probe 16 and the motor 12 to be damaged.
  • the sealing tube 17 is a corrugated tube with a corrugated tube wall.
  • bracket 11 is connected to a displacement system.
  • the displacement system can be a three-dimensional motion module, which is used to drive the entire multi-probe assembly to perform three-dimensional motion.
  • Multiple probes 16 share one holding tank 14 and displacement system. At the same time, the pipetting work does not interfere with each other.
  • Multiple probes 16 of the same specification or multiple probes 16 of different specifications can be configured to improve the efficiency and compatibility of pipetting.
  • a large-sized high-frequency broadband focused ultrasonic transducer is used in the ultrasonic transmitting module to obtain a non-contact ultrasonic pipetting device with high precision and a wide range of adjustable pipetting accuracy: large-sized high-frequency focused ultrasonic transducer
  • the transducer not only has a small focal spot size, but also has a large output sound radiation force, which can realize picoliter ultrasonic pipetting; the high bandwidth of the ultrasonic transducer can ensure that the transducer has a higher frequency range
  • the output of acoustic radiation force realizes that the volume of liquid pipetting at one time can be adjusted in a large range, so that the pipetting accuracy can be adjusted arbitrarily from picoliters to microliters.
  • the ultrasonic excitation component is used to excite each probe 16 to emit ultrasonic waves.
  • the ultrasonic excitation assembly includes a plurality of ultrasonic excitation units, each ultrasonic excitation unit includes a drive circuit and an excitation circuit, wherein the drive circuit is connected to the control circuit, and the excitation circuit is connected to the probe 16, and each ultrasonic excitation unit can work independently.
  • one ultrasonic excitation unit can be connected to one probe 16 to excite the probe 16, or two or more probes 16 can be connected to simultaneously excite the probe 16 correspondingly connected to the ultrasonic excitation unit.
  • Fig. 3 of the specification which is a schematic structural diagram of an ultrasonic excitation component used in the ultrasonic transmitting module of this embodiment.
  • the control circuit controls the drive circuit to generate the required ultrasonic waveform, and then excites the probe 16 through the excitation circuit.
  • the ultrasonic excitation component supports the excitation of two or more probes 16 . Specifically, multiple probes 16 can work independently, or work at the same time or simultaneously. Since the drive circuits and excitation circuits in different ultrasonic excitation units are independent of each other, the excitation parameters of each ultrasonic excitation unit can be adjusted in real time, which can be used to simultaneously excite different probes to emit ultrasonic waves at the same or different frequencies to meet multiple requirements. different pipetting parameters.
  • the excitation parameters include ultrasonic frequency, ultrasonic amplitude, ultrasonic pulse length, pulse repetition frequency, and the like. In this embodiment, one ultrasonic excitation unit can be connected with one probe 16 .
  • the ultrasonic excitation component further includes a power detection unit, configured to detect the energy of the ultrasonic signal emitted by the ultrasonic excitation component, so as to stabilize the amplitude of the ultrasonic excitation signal.
  • the power detection unit includes a signal coupling module and a signal acquisition module.
  • the signal acquisition module measures the attenuated signal of the ultrasonic excitation component coupled by the signal coupling module, thereby obtaining the amplitude of the ultrasonic excitation signal.
  • the control circuit can be realized by FPGA, and can also be realized by MCU such as single-chip microcomputer, ARM processor and DSP.
  • the control circuit is used for ultrasonic excitation control and ultrasonic acquisition processing.
  • the control circuit can also perform mechanical control of the dual-channel probe 16 for moving the spatial position of the probe 16 and the like.
  • Each signal detection channel is connected to a probe 16, and the processing circuits and acquisition circuits of different signal detection channels are independent of each other, so the parameters of each probe 16 can be acquired independently without affecting each other, and can support simultaneous detection of multiple channels.
  • the signal detection component can obtain data such as liquid level height, volume and solution type of the liquid carrier device 2 through the detection signal, and can automatically obtain liquid physical parameters. Quantitative evaluation of liquid physical parameters can be carried out by using parameters such as the amplitude and frequency spectrum of ultrasonic detection signals.
  • a liquid-carrying device 2 When pipetting liquid, a liquid-carrying device 2 is placed above the ultrasonic emission module, and a plurality of regularly arranged liquid-carrying holes are arranged on the liquid-carrying device 2 (generally a porous plate with many small holes regularly arranged, each small hole Contains several liquids that need to be pipetted).
  • the coupling fluid is water.
  • the coupling liquid cooled by the coupling liquid circulation system flows into the multi-probe assembly from the 1 port and 4 port of the holding tank 14 and enters the coupling cavity 13 of the two probes 16, and the coupling liquid fills the coupling cavity 13 and flows from the coupling nozzle
  • the small opening at the top of 15 flows out, and due to the surface tension of the water, a protruding water bag is formed at the opening of the coupling nozzle 15. After the water bag reaches a certain height, it continues to stay along the outer wall of the coupling nozzle 15 and flows into the holding tank 14. , 3 ports flow back to the coupling liquid circulation system.
  • the position of the coupling nozzle 15 corresponds to the position of each probe 16 one by one.
  • the probe 16 is a focused ultrasonic transducer, and its focusing can be physical focusing or lens focusing.
  • the ultrasonic waves emitted by the probe 16 are focused ultrasonic waves. Under the action of the ultrasonic waves, the liquid to be pipetted can fly away from the liquid-carrying device 2 without contact.
  • the distance between the probes 16 is an integral multiple of the pitch of the liquid-carrying holes on the liquid-carrying device 2 , so that the two probes 16 can be aimed at different liquid-carrying holes on the liquid-carrying device 2 at the same time.
  • the displacement system controls the probe 16 of the ultrasonic emitting module to align with the hole on the liquid-carrying device 2 that needs to be pipetted.
  • the two probes 16 each detect the height of the liquid level through the echo, and the high-precision vertical movement of the motor 12 makes the focus of the probe 16 on the liquid level, and the multi-channel excitation component respectively excites the two probes 16 according to the input parameters of the liquid.
  • the liquid in the liquid carrier device 2 flies off the liquid surface under the action of ultrasonic waves and is received by the target liquid carrier plate directly above.
  • the displacement system drives the ultrasonic ultrasonic emission module to move to the next group of pipetting holes, and continues the pipetting operation until all pipetting tasks are completed.
  • the dual-probe ultrasonic pipetting method carried out by the ultrasonic transmitting module of this embodiment when the ultrasonic acoustic parameters (including frequency, amplitude, pulse length and pulse repetition frequency, etc.) are consistent, the dual-probe ultrasonic pipetting can significantly increase the speed of ultrasonic pipetting and efficiency; when the ultrasonic acoustic parameters are inconsistent, the dual-probe ultrasonic pipetting system supports probes 16 with different acoustic parameters to work simultaneously to realize the pipetting function.
  • the ultrasonic acoustic parameters including frequency, amplitude, pulse length and pulse repetition frequency, etc.
  • the ultrasonic pipetting device of the present invention can coordinate the work of a plurality of probes 16 with the same acoustic parameters, and can also support the joint work of a plurality of probes 16 with different acoustic parameters to perform ultrasonic pipetting, and can flexibly control the size of each pipetting, providing high-quality Precision control and pipetting efficiency provide a better solution.
  • the ultrasonic pipetting device of the present invention performs ultrasonic pipetting.
  • the ultrasonic pipetting method includes:
  • Each ultrasonic excitation unit controlling the ultrasonic excitation assembly excites the probe 16 to emit ultrasonic waves
  • the ultrasonic excitation unit excites different probes 16 to emit ultrasonic waves at the same or different frequencies.
  • a kind of ultrasonic pipetting method that adopts the ultrasonic pipetting device that has double probe 16 as described in embodiment 1 to comprise:
  • Each ultrasonic excitation unit controlling the ultrasonic excitation assembly excites the probe 16 to emit ultrasonic waves.
  • the ultrasonic excitation assembly includes a first ultrasonic excitation unit excitation and a second ultrasonic excitation assembly
  • the two probes 16 are respectively a first probe and a second probe
  • the first ultrasonic excitation unit is connected to the first probe
  • the second ultrasonic excitation unit is connected to the second ultrasonic excitation unit.
  • the driving circuits and excitation circuits in different ultrasonic excitation units are independent of each other, so the excitation parameters of each ultrasonic excitation unit can be adjusted independently, so the first ultrasonic excitation unit can excite the first probe to emit ultrasonic waves at the first frequency, and the second The ultrasonic excitation unit excites the second probe to emit ultrasonic waves at the second frequency.
  • the first frequency may be the same as the second frequency, or may be different from the second frequency; when the first frequency is different from the second frequency, the first frequency may be greater than the second frequency, or the first frequency may be less than the second frequency.
  • Frequency so as to excite the two probes 16 of the dual-probe assembly to emit ultrasonic waves at the same or different frequencies.
  • the first frequency is the same as the second frequency, which can significantly improve the speed and efficiency of ultrasonic pipetting.
  • the ultrasonic pipetting device can support probes 16 with different acoustic parameters to work at the same time to realize the pipetting function, so as to be flexibly controlled.
  • the size of each pipette is the same as the second frequency, or may be different from the second frequency; when the first frequency is different from the second frequency, the first frequency may be greater than the second frequency, or the first frequency may be less than the second frequency.
  • the ultrasonic pipetting method of the present invention can be used in an ultrasonic pipetting device with more probes 16 to coordinate multiple probes 16 with the same acoustic parameters, or multiple probes 16 with different acoustic parameters work together to perform ultrasonic pipetting.
  • the present invention provides an ultrasonic pipetting device, which is provided with a plurality of probes for emitting ultrasonic waves to separate the liquid on the liquid-carrying device and fly out vertically to be received by the target liquid-carrying plate directly above it.
  • Each probe is connected to a different ultrasonic excitation unit, and each ultrasonic excitation unit can work independently to support the excitation and setting of various ultrasonic waveform parameters to meet the needs of various excitation parameters.
  • the ultrasonic liquid pipetting device of the present invention can coordinate the work of multiple probes with the same acoustic parameters, and can also support multiple probes with different acoustic parameters to work together for ultrasonic liquid pipetting, and can flexibly control the size of each pipetting liquid, providing high-quality Precision control and pipetting efficiency provide a better solution.
  • each probe is equipped with a motor to drive the probe to move up and down to change the distance between the probe and the liquid surface of the liquid carrier device. Multiple probes can work at the same time and each probe can focus independently. Multiple probes can be flexibly matched. In order to achieve the purpose of providing the use efficiency and applicability of the ultrasonic pipetting device.
  • the ultrasonic pipetting method carried out by the ultrasonic pipetting device of the present invention can coordinate the work of a plurality of probes with the same acoustic parameters to improve the speed and efficiency of ultrasonic pipetting; it can also support the simultaneous work of probes with different acoustic parameters to realize the pipetting function, to flexibly control the size of each pipette.

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  • Health & Medical Sciences (AREA)
  • Clinical Laboratory Science (AREA)
  • Chemical & Material Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Investigating Or Analyzing Materials By The Use Of Ultrasonic Waves (AREA)

Abstract

La présente invention concerne un appareil de pipetage ultrasonore, comprenant un module d'émission ultrasonore et un appareil de transport de liquide ; le module d'émission ultrasonore comprend un constituant multi-sonde, un système de circulation de liquide de couplage et un constituant d'excitation ultrasonore, l'appareil de transport de liquide étant disposé au-dessus du constituant multi-sonde ; le constituant multi-sonde comprend deux sondes ou plus, une cavité de couplage et une buse de couplage, les deux sondes ou plus étant disposées sur le même support, l'appareil de transport de liquide comporte de multiples trous de transport de liquide, le point focal des sondes est aligné avec le niveau de liquide de l'appareil de transport de liquide, la cavité de couplage entoure les sondes, la buse de couplage est disposée entre les sondes et l'appareil de transport de liquide, et la cavité de couplage est remplie intérieurement d'un liquide de couplage ; le système de circulation de liquide de couplage permet au liquide de couplage d'entrer en continu dans la cavité de couplage, et le liquide de couplage déborde de la buse de couplage pour entrer en contact avec la partie inférieure de l'appareil de transport de liquide ; et le constituant d'excitation ultrasonore est connecté à un circuit de commande, et le constituant d'excitation ultrasonore comprend au moins deux unités d'excitation ultrasonore, chaque unité d'excitation ultrasonore étant connectée à une sonde correspondante pour exciter chaque sonde, et les sondes étant utilisées pour émettre des ondes ultrasonores.
PCT/CN2021/109810 2021-07-30 2021-07-30 Appareil et procédé de pipetage ultrasonore Ceased WO2023004795A1 (fr)

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PCT/CN2021/109810 WO2023004795A1 (fr) 2021-07-30 2021-07-30 Appareil et procédé de pipetage ultrasonore

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Application Number Priority Date Filing Date Title
PCT/CN2021/109810 WO2023004795A1 (fr) 2021-07-30 2021-07-30 Appareil et procédé de pipetage ultrasonore

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Citations (3)

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Publication number Priority date Publication date Assignee Title
US10325768B1 (en) * 2015-09-03 2019-06-18 Labcyte Inc. Focused acoustic radiation for rapid sequential ejection of subwavelength droplets
CN111495455A (zh) * 2020-05-09 2020-08-07 中国科学院深圳先进技术研究院 非接触式超声移液装置及方法
CN113070106A (zh) * 2020-01-05 2021-07-06 天津大学 一种超声移液装置及基于相控阵技术实现超声移液的方法

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US10325768B1 (en) * 2015-09-03 2019-06-18 Labcyte Inc. Focused acoustic radiation for rapid sequential ejection of subwavelength droplets
CN113070106A (zh) * 2020-01-05 2021-07-06 天津大学 一种超声移液装置及基于相控阵技术实现超声移液的方法
CN111495455A (zh) * 2020-05-09 2020-08-07 中国科学院深圳先进技术研究院 非接触式超声移液装置及方法

Non-Patent Citations (1)

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ANONYMOUS: "Beckman Coulter Echo nano upgrade pipetting system - a new generation of pipetting technology", XP093029732, Retrieved from the Internet <URL:https://v.qq.com/x/page/j3014q99imd.html> [retrieved on 20230307] *

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