An ultrasonic phased array multi-droplet automatic manipulation and detection system and method

Abstract

The application discloses an ultrasonic phased array multi-droplet automatic control and detection system and method, which comprises an ultrasonic phased array driving device, an optical flat, a phased array circuit board and a droplet detection platform which are fixed and installed on the optical flat in sequence from top to bottom; the droplet detection platform comprises a super-hydrophobic surface; the phased array circuit board is used for capturing droplets on the super-hydrophobic surface and moving the droplets to a required position; a multi-droplet automatic titration device is fixed and installed on the optical flat and is used for generating droplets with a required volume; the ultrasonic phased array driving device further comprises an acquisition module which is used for acquiring droplet related information to perform droplet detection; a plurality of acoustic field focal points are generated by a computer to realize non-contact control of a plurality of droplets; the droplets are generated in an automatic control mode, the accuracy of droplet control is improved, and high-throughput detection of droplet samples is realized by cooperating with a fluorescence detection sensor and a biological probe.

Description

TECHNICAL FIELD

[0001] The present application relates to multi-droplet generation and multi-droplet manipulation technology, in particular to an ultrasonic phased array multi-droplet automatic manipulation and detection system and method. BACKGROUND

[0002] Droplet manipulation technology refers to the precise control of the movement, merging, splitting or arrangement of droplets in the internal or external environment through external control means (such as electric field, magnetic field, optical field, acoustic field, etc.). The goal of droplet manipulation usually includes precise control of the distribution, size, shape and interaction of droplets, and is widely used in medical diagnosis, chemical synthesis, single molecule experiment and other fields. Internal droplet manipulation refers to the manipulation of droplets in a microfluidic device. However, this method has the problems of uneven distribution of droplets, limited operation precision, high cost due to complex equipment, and directly affects the high-throughput biochemical detection rate of droplets. External droplet manipulation does not depend on the fluid dynamics in the microfluidic channel and the microchannel, and mainly manipulates the droplets through physical or chemical means. This kind of technology is suitable for macro or larger scale droplet operation, and usually uses external physical field or chemical environment to change the shape, position, size or motion state of the droplet. This technology does not involve fine flow control in microfluidics, but uses more direct external intervention. It is more suitable for high-throughput biochemical detection.

[0003] Electric field as one of the droplet manipulation technologies, needs high voltage in the driving process, and the equipment is relatively large in size; magnetic field is used to manipulate magnetic droplets, and the added magnetic material may affect the subsequent chemical detection results; light field changes the phototaxis of droplets to realize droplet control, and the heat generated under the action of light will damage the sample to be tested.

[0004] Acoustic field as one of the droplet manipulation technologies is widely used, and its basic principle is: when the distance between the ultrasonic source and the target to be tested is an integer multiple of half the wavelength, under the action of the radiated sound pressure, the acoustic radiation force parallel to the ultrasonic source is generated. If the sound source is placed above the droplet, the acoustic radiation force can offset part of the effect of gravity. If the acoustic radiation force is greater than or equal to the gravity, the droplet is suspended, and the position of the droplet is changed by dragging the position of the sound source.

[0005] The current droplet operation and detection need droplets which are mostly obtained by manual titration with a pipette. If different volumes of droplets need to be titrated at different positions of the detection platform, the parameters of the pipette need to be adjusted repeatedly. If the types of droplets are different, the original solution needs to be discharged and another solution needs to be sucked. This leads to a decrease in the detection rate of droplets, and long-term use of the pipette will cause damage to the joints of the human hand. SUMMARY

[0006] The purpose of the present application is to solve the problems in the prior art.

[0007] The technical scheme adopted by the present application to solve its technical problems is to provide an ultrasonic phased array multi-droplet automatic operation and detection system, comprising:

[0008] An ultrasonic phased array driving device, comprising an optical flat made of metal and a phased array circuit board and a droplet detection platform fixedly installed on the optical flat in sequence from top to bottom; the droplet detection platform comprises a super-hydrophobic surface, and the phased array circuit board is used for capturing droplets on the super-hydrophobic surface and moving the droplets to a desired position;

[0009] A multi-droplet automatic titration device fixedly installed on the optical flat; the multi-droplet automatic titration device is used for generating droplets with a desired volume and titrating the droplets on the super-hydrophobic surface;

[0010] A waste liquid recovery device fixedly installed beside the droplet detection platform on the optical flat, used for capturing and recovering droplets on the droplet detection platform after detection is completed;

[0011] A communication module connected with the upper computer, the ultrasonic phased array driving device and the multi-droplet automatic titration device to realize communication between the upper computer and the ultrasonic phased array driving device and communication between the upper computer and the multi-droplet automatic titration device;

[0012] The ultrasonic phased array driving device further comprises an acquisition module used for acquiring droplet-related information and sending the information to the upper computer for droplet detection.

[0013] Preferably, the circuit arranged on the phased array circuit board comprises:

[0014] A microcontroller used for generating a control sequence according to an instruction of the upper computer and decomposing the control sequence into a plurality of clock signals and outputting the clock signals through a plurality of output ends respectively;

[0015] A plurality of shift registers, an input end of each shift register is connected with an output end of the microcontroller, and a plurality of output ends of each shift register output a plurality of parallel signals;

[0016] A plurality of drive amplifiers, an input end of each drive amplifier is connected with an output end of the shift register, and an output end of each drive amplifier outputs an amplified signal;

[0017] A plurality of ultrasonic wave generators, an input end of each ultrasonic wave generator is connected with an output end of the drive amplifier, and an output end of each ultrasonic wave generator generates ultrasonic waves; the ultrasonic waves generated by the plurality of ultrasonic wave generators are focused to form a plurality of acoustic field focal points to control a plurality of droplets respectively.

[0018] Preferably, the microcontroller is an FPGA chip, comprising at least 32 output ends used for outputting clock signals; each output end is electrically connected with an input end of 1 shift register.

[0019] The shift register has 32, each shift register includes 1 input and 8 output, each output is electrically connected with the input of a driver amplifier;

[0020] The driver amplifier has 128, each driver amplifier includes 2 input and 2 output, each output is electrically connected with the positive input of two ultrasonic generators;

[0021] The ultrasonic generator has 256, 16x16 array distribution; Each ultrasonic generator includes 2 input and 1 output, the negative input is grounded, the output generates ultrasonic according to the input signal.

[0022] Preferably, the multi-droplet automatic titration device comprises:

[0023] Liquid storage module for storing titration solution;

[0024] Titration module for generating droplets;

[0025] Droplet pumping module for pumping titration solution to titration module; the input is connected with the liquid storage module, and the output is connected with the titration module;

[0026] The first driving circuit is connected with the titration module and the droplet pumping module respectively for power supply.

[0027] Preferably, the liquid storage module comprises a plurality of cryogenic tubes, the cryogenic tubes are fixed on the optical flat through the cryogenic tube support; a plurality of through holes are reserved on the cryogenic tube support to correspond to the placement of a plurality of cryogenic tubes, and the cryogenic tube support is fixed on the optical flat through screws;

[0028] The titration module comprises a servo motor, a servo motor connecting rod, a plurality of PP adapters and a plurality of dispensing needles; the servo motor connecting rod comprises a horizontal bar and two feet extending from the two ends of the horizontal bar perpendicular to the horizontal bar, the two feet are parallel and equal in length; the servo motor is fixed on a metal plate through a motor support, the metal plate is fixed on the optical flat through a metal plate support and located above the cryogenic tube; a servo motor arm is installed on the output shaft of the servo motor, the servo motor arm connects one of the feet of the servo motor connecting rod, the servo motor connecting rod is fixedly connected with an adapter support, and the adapter support is provided with a plurality of through holes; each PP adapter is composed of an inverted taper adapter and a luer male head, the inverted taper adapter and the luer male head are located at the two ends of the through hole of the adapter support respectively, one end of the luer male head is screwed with the inverted taper adapter to be fixed on the adapter support, and the other end of the luer male head is installed with a dispensing needle for generating droplets.

[0029] Preferably, two limiting crossbars are extended outwardly on the two feet of the servo motor connecting rod in the direction parallel to the crossbar, the two limiting crossbars pass through the slot holes of the two limiting supports respectively, and the two limiting supports are fixed on the metal plate; one of the limiting supports is provided with a bearing round hole, a bearing is installed in the bearing round hole, and a bearing crossbar is further extended on the foot connected with the limiting support and passes through the middle of the bearing.

[0030] Preferably, the collection module comprises a fluorescence detector and an industrial camera fixedly installed on the edge of the optical flat; the camera of the industrial camera has a field of view angle of up to 120° and is fixedly installed on the optical flat in a lens-up mode below the super-hydrophobic surface of the droplet detection platform.

[0031] Preferably, the waste liquid recovery device comprises:

[0032] a second liquid storage module for storing the recovered droplets;

[0033] a second droplet pumping module for pumping the droplets captured by the recovery module to the second liquid storage module; the input end is connected with the output end of the recovery module, and the output end is connected with the second liquid storage module;

[0034] a recovery module, the input end of which is flush with the super-hydrophobic surface of the droplet detection platform, and the output end of which is connected with the second droplet pumping module;

[0035] a second driving circuit connected with the second droplet pumping module and the recovery module for power supply.

[0036] Preferably, the second liquid storage module comprises a freezing tube fixed on the optical flat through a recovery support; the recovery support is provided with a through hole for placing the freezing tube and is fixed on the optical flat through a screw; the second droplet pumping module comprises a micro peristaltic pump, the positive and negative power supply lines of which are connected with the second driving circuit, a BPT hose is sleeved on the rotor of the micro peristaltic pump, one end of the BPT hose is connected with the freezing tube of the second liquid storage module, and the other end of the BPT hose is connected with an adapter of the recovery module; the recovery module comprises a PP adapter and a dispensing needle, the dispensing needle is connected with the BPT hose of the second droplet pumping module through the PP adapter; the dispensing needle is flush with the super-hydrophobic surface of the droplet detection platform.

[0037] The number of freezing tubes, the number of micro peristaltic pumps, the number of PP adapters and the number of dispensing needles are consistent, a plurality of freezing tubes are connected with one end of a plurality of BPT hoses one by one, the other end of the plurality of BPT hoses is connected with a plurality of PP adapters one by one, and one dispensing needle is fixedly installed at the end of each PP adapter.

[0038] The application also provides an ultrasonic phased array multi-droplet automatic control and detection method, which adopts the system described in any of the above and comprises the following steps:

[0039] The multi-droplet automatic titration device is started, and multiple droplets with a volume meeting the expected requirements are generated on the droplet detection platform;

[0040] The current positions of the multiple droplets are detected by using the acquisition module, and the phased array circuit board is started to generate multiple acoustic field focal points to capture the multiple droplets;

[0041] According to the preset end position of the droplets, the shortest paths of the multiple droplets moving to the end position are calculated, and the multiple acoustic field focal points are controlled to move the multiple droplets according to the shortest paths;

[0042] After the multiple droplets all reach the target detection position, the acquisition module is used to collect droplet related information and upload the information to the upper computer, and the upper computer analyzes the droplet composition according to the related information uploaded by the acquisition module;

[0043] After the upper computer completes the analysis of the droplet composition, the multiple droplets are moved to the recycling area by the multiple acoustic field focal points generated by the phased array circuit board;

[0044] The second droplet pumping module of the recycling device is started, and the droplets are recycled and stored in the second liquid storage module.

[0045] The application has the following beneficial effects:

[0046] (1) The application can control the ultrasonic phased array to generate multiple acoustic field focal points to capture and simultaneously move multiple droplets without contacting the droplets, and the ultrasonic phased array control droplet method significantly improves the distance of the controllable droplets;

[0047] (2) The automatic titration device of the droplets is introduced, the generation of the controlled droplets is automated, and the waste liquid recycling system is added to avoid the pollution of the droplets in the detection process;

[0048] (3) The fluorescence detection module is introduced to facilitate the acquisition of the droplet sample composition information. The whole application realizes automatic operation and analysis, reduces human intervention, and improves the droplet control efficiency.

[0049] The application will be further described in detail in combination with the drawings and embodiments, but the application is not limited to the embodiments. BRIEF DESCRIPTION OF DRAWINGS

[0050] Figure 1 The figure is a system structure schematic diagram of the embodiment of the application;

[0051] Figure 2 The figure is an exploded and overall installation diagram of the embodiment of the application;

[0052] Figure 3 exploded and overall installation view of the ultrasonic phased array driving device;

[0053] Figure 4 connection schematic diagram of the ultrasonic phased array driving device;

[0054] Figure 5 exploded and overall installation view of the multi-droplet automatic titration device;

[0055] Figure 6 upper metal plate design diagram of the multi-droplet automatic titration device;

[0056] Figure 7 first driving circuit connection diagram of the multi-droplet automatic titration device;

[0057] Figure 8 first driving circuit principle diagram of the multi-droplet automatic titration device;

[0058] Figure 9 exploded and overall installation view of the waste liquid recovery device;

[0059] Figure 10 flow chart of the method of the embodiment of the present application.

[0060] Marked for explanation: 100-ultrasonic phased array driving device, 101-phased array circuit board, 102-phased array circuit fixed metal plate, 103-TDT-2020 aluminum profile-250mm, 104-2020 angle code, 105-TDT-2020 aluminum profile-300mm, 106-droplet detection platform, 107-industrial camera, 108-optical flat, 109-fluorescence detection module support, 110-fluorescence detector; 200-multi-droplet automatic titration device, 201-first driving circuit, 202-TDT-2020 aluminum profile-135mm, 203-peristaltic pump pressing plate, 204-limiting support(with bearing hole), 205-bearing(606ZZ 6*17*6), 206-upper metal plate, 207-first micro peristaltic pump, 208-first frozen tube, 209-frozen tube support, 210-limiting support(without bearing hole), 211-servo motor support, 212-servo motor arm, 213-servo motor arm, 214-servo motor connecting rod, 215-first luer male head, 216-adaptor support, 217-first inverted taper joint; 300-waste liquid recovery device, 301-second frozen tube, 302-waste liquid recovery support, 303-second luer male head, 304-second inverted taper joint, 305-second micro peristaltic pump. DETAILED DESCRIPTION

[0061] Reference Figure 1 and Figure 2As shown, the system structure diagram of the embodiment of the application is decomposed and the overall installation diagram, the system comprises:

[0062] The ultrasonic phased array driving device 100 comprises an optical flat 108, a phased array circuit board 101 and a droplet detection platform 103 fixed and installed on the optical flat 108 in sequence from top to bottom; the droplet detection platform 106 comprises a super-hydrophobic surface, the phased array circuit board 101 is used for capturing the droplet on the super-hydrophobic surface and moving the droplet to a required position; the ultrasonic phased array driving device 100 further comprises a collection module used for collecting droplet related information to perform droplet detection; the collection module comprises an industrial camera 107 and a fluorescence detection device 110;

[0063] The multi-droplet automatic titration device 200 is fixed and installed on the optical flat 108 and is used for generating a droplet with a required volume;

[0064] The waste liquid recovery device 300 is fixed and installed beside the droplet detection platform 106 on the optical flat 108 and is used for capturing and recovering the droplet on the droplet detection platform 106 after detection; the waste liquid recovery device 300 comprises a second liquid storage module used for storing the recovered droplet, a second droplet pumping module with an input end connected with the recovery module and an output end connected with the second liquid storage module, a recovery module with an input end flush with the super-hydrophobic surface of the droplet detection platform 106 and an output end connected with the second droplet pumping module, and a second driving circuit connected with the second droplet pumping module and the recovery module to supply power;

[0065] The auxiliary support metal piece comprises an aluminum profile, the optical flat 108, a metal corner code and a phased array circuit fixed metal plate.

[0066] Specifically, the ultrasonic phased array driving device 100 comprises a phased array circuit fixed metal plate 102, the phased array circuit fixed metal plate 102 is fixed on the optical flat 108 through a column structure composed of an aluminum profile, a beam structure composed of an aluminum profile and a metal corner code. The multi-droplet automatic titration device 200 comprises an upper metal plate 206, the upper metal plate 206 is fixed on the optical flat 108 through a column structure composed of an aluminum profile.

[0067] Referring to Figure 3As shown, the ultrasonic phased array driving device 100 includes a bottom optical flat 108, four 300mm aluminum profiles 105 are fixed on the optical flat to form a column, two 250mm aluminum profiles 103 form a beam structure fixed by angle code 104. The phased array circuit fixed metal plate is fixed on the beam, and the phased array circuit board 101 is placed on the metal sheet and fixed by screws and nuts. The industrial camera 107 is fixed in the center of the optical flat 108 with the lens facing up. The droplet detection platform 106 is placed directly below the phased array circuit board 101 and fixed on the optical flat 108. The fluorescence detection module includes a fluorescence detector 110 and a fluorescence detection module support 109. The fluorescence detection module support 109 is fixed on the edge of the optical flat 108 by screws, and two sets of fluorescence detectors 110 are fixed on the fluorescence detection module support 109 by screws and bolts. The end of the fluorescence detection module support 109 has a certain inclination angle, which is beneficial to the irradiation of the light beam on the hydrophobic surface.

[0068] Specifically, the phased array circuit board 101 is an ultrasonic phased array PCB circuit board, which includes a main control module including an integrated main control chip and a communication module, a signal processing module connected to the signal output end of the main control module, and an ultrasonic wave generator connected to the output end of the signal processing module. The above-mentioned modules are connected by electrical connection. The PCB circuit board is placed on a metal sheet and fixed by screws.

[0069] The ultrasonic wave generator used in the embodiment of the application has a center frequency of 40kHz and is arranged in an array in a 16x16 manner with a spacing of 1mm between adjacent probes. The main control chip used is an FPGA CoreEP4CE6, which is used to generate a clock control sequence, and the communication module is an Arduino Nano. The signal processing module includes a 74HC595D shift register and a MIC4127YME drive amplifier. The signal processing module converts the serial clock signal generated by the FPGA into eight parallel output signals (amplitude 5V) acting on the input end of the drive amplifier, and the drive amplifier amplifies the input signal into a pulse signal (amplitude 6V-15V) with the same input voltage amplitude. By adjusting the amplitude and phase difference of the output pulse signal, the focus position of the sound field can be changed, and then the movement of the droplets can be captured.

[0070] The ultrasonic phased array PCB circuit board designed in the embodiment of the application includes one main control module, 32 shift registers, 128 drive amplifiers, and 256 ultrasonic wave generators. The ultrasonic wave generators are arranged in a 16x16 manner. Referring to Figure 4As shown, one of the topologies is exemplified: when the FPGA receives the host computer instruction, a clock signal is generated according to the instruction, the clock signal is input into the shift register, the clock signal is output in parallel and acts on the driving amplifier. The driving amplifier amplifies the input signal and acts on the ultrasonic generator to generate a sound wave signal with sufficient amplitude. By adjusting the clock signal trigger time, the phase of the sound wave signal generated by the final ultrasonic generator is realized, thereby realizing the generation of the sound field focal point position change.

[0071] Specifically, the droplet detection platform 106 is composed of a platform support and a super-hydrophobic surface. The platform support is a hollow structure and is made by integral molding. The platform support is placed on the optical flat 108 and is located 16 cm below the phased array circuit board 101. The super-hydrophobic surface is made by spraying a hydrophobic material (XN-204L hydrophobic coating) on a glass substrate, heating at 150°C for 1 hour and then cooling. The super-hydrophobic surface is placed flat on the center of the platform support and serves as a droplet motion control platform.

[0072] Specifically, the industrial camera 107 is fixed at the center of the optical flat 108 and is used to collect droplet motion images. During use, the industrial camera 107 sends droplet image information to the host computer at fixed time intervals to realize the feedback function. The host computer judges whether the droplet is successfully captured according to the image information, compares the expected preset position of the droplet with the actual position, and corrects the droplet motion trajectory.

[0073] Specifically, the fluorescence detection module is composed of two fluorescence detectors 110 and a fluorescence detection module support 109. The fluorescence detectors 110 are fixed on the fluorescence detection module support 109 by bolts. Since the support itself has a certain inclination angle, the light beams emitted by the fluorescence detectors 110 can better irradiate the hydrophobic surface. During use, the fluorescence detection module sends droplet composition information to the host computer in the form of an electrical signal for detection and analysis. The specific principle is as follows: the fluorescence detection module emits laser light, which irradiates the measured sample (i.e., the droplet). The measured sample reflects a certain fluorescence wavelength. After the fluorescence is filtered, it is focused on the photodetector integrated inside by the fluorescence collection lens. After photoelectric conversion and amplification processing, the corresponding electrical signal is obtained. Finally, the electrical signal is sent to the host computer through serial communication. This information transmission is uninterrupted and at certain time intervals. The host computer analyzes these electrical signals and thus obtains the droplet composition. The model of the fluorescence detector used in the embodiment of the present application is MNS-MFD-365.

[0074] Referring to Figure 5As shown, it is the exploded and overall installation view of the multi-droplet automatic titration device 200, which can be divided into a first driving circuit, a droplet pumping module, a liquid storage module, a titration module, and auxiliary metal parts; the titration module further comprises a servo motor. The multi-droplet automatic titration device 200 is fixed on the optical flat 108 as a whole, and the auxiliary metal parts used include six aluminum profiles 202 (TDT-2020 aluminum profile-135mm) and an upper metal plate 206 (specification reference Figure 6 ).

[0075] The droplet pumping device is composed of four first micro peristaltic pumps 207. In the embodiment of the application, the micro peristaltic pump with the model LFP101ADB is adopted, the rated flow is 0.1ml / min, and the rated input voltage is 6V. The four first micro peristaltic pumps 207 are respectively placed in the holes with a diameter of 19mm in the upper metal plate 206 and fixed by the peristaltic pump pressing plate 203 and screws.

[0076] The liquid storage module is composed of four 5ml first cryogenic tubes 208 and a cryogenic tube support 209. The lid of each first cryogenic tube 208 is pre-provided with a hole with a diameter of 1.5mm-2mm. The four first cryogenic tubes 208 can respectively store different kinds of solutions to be titrated and are placed in the cryogenic tube support 209. The cryogenic tube support 209 is integrally formed and contains four groove holes with a diameter of 16.4mm, which facilitates the placement and removal of the cryogenic tubes. The support base is pre-provided with four fixing holes, which are fixed on the edge of the optical flat 108 by screws.

[0077] The titration module comprises a servo motor (TD8115), a servo motor support 211, a servo motor connecting rod 214, a limiting support (with bearing side 204 and without bearing side 210), a bearing 205 (606ZZ, 6*17*6), a first PP adapter, an adapter support 214 and a dispensing needle. Reference Figure 7 The rotor of the first micro peristaltic pump 207 is sleeved with a BPT hose, wherein the liquid output end is connected with the first inverted taper joint 217 of the titration module, and the liquid input end is connected with the first cryogenic tube 208 and enters the inside of the first cryogenic tube 208 through the hole pre-provided in the lid of the first cryogenic tube 208. The positive and negative leads of the first micro peristaltic pump 207 are respectively connected with the positive and negative terminals of the peristaltic pump of the first driving circuit.

[0078] The servo motor is fixed on the upper metal plate 206 by screws and nuts, specifically fixed on the inner diameter 3.2mm hole of the upper metal plate 206 through the servo motor support 211, the servo motor output shaft is connected with the servo motor arms 212 and 213, and the servo motor arms 212 and 213 are connected with the servo motor connecting rod 214. The limiting support is divided into bearing side and non-bearing side, the bearing side is installed on the length 61mm slot hole of the upper metal plate, the bearing is installed in the bearing hole of the limiting support 204, and the non-bearing side is installed in the length 64mm slot hole of the upper metal plate. The rod extended from the servo motor connecting rod 214 respectively passes through the slot hole reserved in the bearing 205 and the limiting support 204. The adapter support 216 is installed in the slot hole at the end of the servo motor connecting rod 214, and the installation position is adjusted by adapting the height of the liquid drop detection platform 106. The first PP adapter is divided into two parts of the first inverted taper adapter 217 and the first luer male head 215, wherein the liquid output end of the first micro peristaltic pump 207 is connected with the first inverted taper adapter 217. The first inverted taper adapter 217 passes through the upper part of the adapter support 216, the first luer male head 215 passes through the lower part of the adapter support 216, and the two parts are tightly fixed; the end of the first luer male head 215 is fixed with a point glue needle head.

[0079] The first driving circuit 201 is fixed on the upper metal plate 206 by the aluminum profile 202. The first driving circuit includes a core module (the main control chip is Arduino nano), two power conversion chips (TPS5450), four switching devices (S9012 and JQC-3FF-005-S-Z relay), and input and output terminals. Referring to Figure 7 and Figure 8 The first driving circuit connection diagram and principle diagram of the multi-liquid drop automatic titration device; the main control chip generates I / O signals and interacts with the communication module data. Two power conversion chips are used to stabilize the external input voltage at 5V and 6V. The 5V voltage is used to power the relay, and the 6V voltage is used to power the servo motor and pumping device in the titration module. The peristaltic pump terminal adopts KF301-2PIN terminal, the communication interface adopts KF301-3PIN terminal, and the servo motor interface adopts 3PIN row needle with a pitch of 2.54mm. When the host computer issues a serial port command, the microprocessor controls the relay switch by outputting a level signal with a corresponding time, and then controls the opening and closing time of the peristaltic pump. During the titration process, the microprocessor outputs a pulse signal to the servo motor, so that the servo motor connecting rod remains parallel to the working platform; when the titration operation is completed, the microprocessor outputs a pulse signal again to place the servo motor connecting rod in a vertical state, so as to avoid the blocking of the adapter pipe support affecting the generation of the sound field focal point.

[0080] Referring to Figure 9As shown, the waste liquid recovery device includes two second micro peristaltic pumps 305, a BPT hose, a PP adapter, a recovery device support 302, and two sets of 5ml second cryogenic tubes 301. The recovery device support 302 is fixed on the optical flat 108 by screws. The PP adapter includes a second inverted taper joint 304 and a second luer male connector 303, and the end of the second luer male connector 303 is connected to a dispensing needle. The PP adapter is placed in the fixed slot of the recovery device support 302, and the dispensing needle is flush with the hydrophobic surface after installation. The BPT hose is sleeved on the rotor of the second micro peristaltic pump 305, and the input end is connected to the second inverted taper joint 304 and the output end is connected to the second cryogenic tube 301. The second micro peristaltic pump 305 is placed in the upper hole of the recovery device support 302. The cryogenic tube is placed in the lower hole. After the droplet detection is completed, the acoustic focus is operated to move the droplet to the vicinity of the dispensing needle of the recovery device, and the power supply of the second micro peristaltic pump 305 is turned on at this time to pump the droplet back into the second cryogenic tube 301.

[0081] Referring to Figure 10 As shown, when multiple droplets are controlled and detected using the embodiment of the present application, the micro peristaltic pump of the titration module works to generate droplets on the droplet detection platform until the volume of the droplets meets the expected requirements. After the droplets are generated, the servo motor works so that the servo motor connecting rod is in a vertical state. Then the current position of the multiple droplets is detected in real time by visual detection, and there are many such algorithms, and the embodiment of the present application uses a droplet target detection method based on deep learning; the phased array circuit is turned on to generate multiple acoustic foci to capture multiple droplets. After the droplets are successfully captured, since the end position of the droplet movement is set in advance, the A* algorithm (8 directions) can be used to generate the shortest path of the multiple droplets to the end position, and then the multiple acoustic foci are controlled to move according to the shortest path. During the movement, the visual feedback method is used to judge whether the droplets are successfully captured, that is, whether the difference between the droplet position coordinates and the acoustic focus position coordinates is kept within 10 pixels, if yes, it means that the droplets are successfully captured, otherwise, it means that the capture fails, and the current droplet position is returned to re-capture and sent to the target detection position. After the droplets reach the target detection position, the information collected by the fluorescence detection module is used to detect the composition of the droplets, and the wavelengths of the fluorescent light reflected by different substances are different, so the electric signals obtained through the photoelectric transducer are also different. The electric signals are continuously sent back to the upper computer at a certain time interval and displayed, and the composition of the droplets is judged according to the size of the electric signals. After the detection is completed, the droplets are moved to the recovery area, the power supply of the peristaltic pump of the recovery device is turned on, and the droplets are recovered.

[0082] The application is an ultrasonic phased array multi-droplet automatic control device, which can control the ultrasonic phased array to generate multiple acoustic field focal points to capture and operate multiple droplets to move simultaneously without contacting the droplets. The ultrasonic phased array control droplet method can significantly improve the distance of the controlled droplets. The automatic titration device is introduced, the generation of the controlled droplets is automated, the fluorescence detection module is introduced to facilitate the acquisition of droplet sample composition information. The waste liquid recovery system is added to avoid droplet pollution during the detection process. The complete application realizes automatic operation and analysis, reduces human intervention, and improves the efficiency of droplet control.

[0083] It can be seen that the application can realize non-contact control of multiple droplets by generating multiple acoustic field focal points through a computer. In addition, the generation of the controlled droplets is generated in an automatic control mode, replacing the traditional manual operation mode, improving the accuracy of droplet control, and cooperating with the fluorescence detection sensor and the biological probe to realize high-throughput detection of the droplet sample.

[0084] The above is only a preferred embodiment of the application and is not intended to limit the application. Any modification, equivalent replacement, improvement, etc. within the spirit and principles of the application shall be included in the protection scope of the application.

Claims

1. An ultrasonic phased array multi-droplet automatic manipulation and detection system, characterized in that, The application relates to an ultrasonic phased array driving device, a multi-droplet automatic titration device, a waste liquid recovery device and a communication module. The ultrasonic phased array driving device comprises an optical flat made of metal, a phased array circuit board and a droplet detection platform which are fixedly installed on the optical flat in sequence from top to bottom; the droplet detection platform comprises a super-hydrophobic surface, the phased array circuit board is provided with an ultrasonic wave generator, and the ultrasonic wave generator is used for capturing liquid droplets on the super-hydrophobic surface and moving the liquid droplets to a required position. The multi-droplet automatic titration device is fixedly installed on the optical flat; the multi-droplet automatic titration device is used for generating liquid droplets with a required volume and titrating the liquid droplets on the super-hydrophobic surface. The waste liquid recovery device is fixedly installed beside the droplet detection platform on the optical flat and is used for capturing and recovering the liquid droplets on the droplet detection platform after detection is completed. The communication module is connected with an upper computer, the ultrasonic phased array driving device and the multi-droplet automatic titration device to realize communication between the upper computer and the ultrasonic phased array driving device and communication between the upper computer and the multi-droplet automatic titration device. The ultrasonic phased array driving device further comprises an acquisition module which is used for acquiring droplet related information and sending the droplet related information to the upper computer to detect the droplets. The multi-droplet automatic titration device comprises a liquid storage module, a titration module, a droplet pumping module, a first driving circuit and a second driving circuit. The liquid storage module is used for storing titration solution. The titration module is used for generating liquid droplets. The droplet pumping module is used for pumping the titration solution to the titration module; an input end of the droplet pumping module is connected with the liquid storage module, and an output end of the droplet pumping module is connected with the titration module. The first driving circuit is connected with the titration module and the droplet pumping module respectively to supply power. The liquid storage module comprises a plurality of freezing tubes which are fixed on the optical flat through a freezing tube support; a plurality of through holes are reserved on the freezing tube support to correspond to the placement of the freezing tubes, and the freezing tube support is fixed on the optical flat through screws. The titration module comprises a servo motor, a servo motor connecting rod, a plurality of PP adapters and a plurality of dispensing needles; the servo motor connecting rod comprises a horizontal rod and two feet which extend in a direction perpendicular to the horizontal rod from two ends of the horizontal rod, and the two feet are parallel and have equal lengths; the servo motor is fixed on a metal plate through a motor support, the metal plate is fixed on the optical flat through a metal plate support and is located above the freezing tubes; a servo motor arm is installed on an output shaft of the servo motor, the servo motor arm is connected with one of the feet of the servo motor connecting rod, an adapter support is fixedly connected on the servo motor connecting rod, and a plurality of through holes are arranged on the adapter support; each PP adapter is composed of an inverted taper adapter and a luer male head, the inverted taper adapter and the luer male head are respectively located at two ends of the through hole of the adapter support, one end of the luer male head is screwed with the inverted taper adapter to be fixed on the adapter support, and the other end of the luer male head is installed with a dispensing needle to generate liquid droplets. Two limiting horizontal rods are extended outward from the two feet of the servo motor connecting rod in a direction parallel to the horizontal rod, two limiting supports are fixed on the metal plate, and the two limiting horizontal rods respectively pass through the slot holes of the two limiting supports; one of the limiting supports is provided with a bearing round hole, a bearing is installed in the bearing round hole, and a bearing horizontal rod is further extended from the foot connected with the limiting support.

2. The ultrasonic phased array multi-droplet automatic manipulation and detection system according to claim 1, characterized in that, The phased array circuit board is provided with a circuit. A microcontroller is configured to generate a control sequence according to an instruction of an upper computer and to divide the control sequence into a plurality of clock signals, and output the clock signals through a plurality of output terminals, respectively. A plurality of shift registers are connected to the output terminals of the microcontroller, respectively, and output a plurality of parallel signals through a plurality of output terminals. A plurality of drive amplifiers are connected to the output terminals of the shift registers, respectively, and output amplified signals through output terminals. A plurality of ultrasonic generators are connected to the output terminals of the drive amplifiers, respectively, and generate ultrasonic waves through output terminals, and the ultrasonic waves generated by the plurality of ultrasonic generators are focused to form a plurality of acoustic field focal points to control a plurality of liquid droplets, respectively.

3. The ultrasonic phased array multi-droplet automatic manipulation and detection system according to claim 2, wherein, The microcontroller is an FPGA chip, and includes at least 32 output terminals configured to output clock signals. Each of the 32 shift registers includes one input terminal and eight output terminals, and each of the output terminals is electrically connected to one input terminal of one drive amplifier. Each of the 128 drive amplifiers includes two input terminals and two output terminals, and each of the output terminals is electrically connected to the positive input terminals of two ultrasonic generators. The 256 ultrasonic generators are arranged in a 16×16 array, and each of the ultrasonic generators includes two input terminals and one output terminal, and the negative input terminal is grounded, and the output terminal generates ultrasonic waves according to the input signals.

4. The ultrasonic phased array multi-droplet automatic manipulation and detection system according to claim 1, wherein, The collection module includes a fluorescence detector and an industrial camera fixedly installed on the edge of the optical flat.

5. The ultrasonic phased array multi-droplet automated manipulation and detection system of claim 1, wherein, The camera of the industrial camera has a field of view of up to 120°, and is fixedly installed on the optical flat in a lens-up manner below the super-hydrophobic surface of the liquid droplet detection platform. The waste liquid recovery device includes: A second liquid storage module configured to store the recovered liquid droplets. A second liquid droplet pumping module configured to pump the liquid droplets captured by the recovery module to the second liquid storage module, and having an input terminal connected to the output terminal of the recovery module and an output terminal connected to the second liquid storage module. The recovery module has an input terminal flush with the super-hydrophobic surface of the liquid droplet detection platform and an output terminal connected to the second liquid droplet pumping module.

6. The ultrasonic phased array multi-droplet automatic manipulation and detection system according to claim 5, wherein, A second driving circuit connected to the second liquid droplet pumping module and the recovery module for power supply. The second liquid storage module includes a freezing tube fixed on the optical flat by a recovery bracket, and the recovery bracket has a through hole for placing the freezing tube and is fixed on the optical flat by a screw. The second liquid droplet pumping module includes a micro peristaltic pump, and the positive and negative power supply lines of the micro peristaltic pump are connected to the second driving circuit. The rotor of the micro peristaltic pump is sleeved with a BPT hose, one end of the BPT hose is connected to the freezing tube of the second liquid storage module, and the other end of the BPT hose is connected to an adapter of the recovery module. The recovery module includes a PP adapter and a dispensing needle, and the dispensing needle is connected to the BPT hose of the second liquid droplet pumping module through the PP adapter. The dispensing needle is flush with the super-hydrophobic surface of the liquid droplet detection platform. The number of the freezing tubes, the number of the micro peristaltic pumps, the number of the PP adapters and the number of the dispensing needles are consistent, a plurality of freezing tubes are connected with one end of a plurality of BPT hoses one by one, the other end of the plurality of BPT hoses is connected with the plurality of PP adapters one by one, and one dispensing needle is fixedly installed at the end of each PP adapter.

7. An ultrasonic phased array multi-droplet automatic manipulation and detection method, using the system of any one of claims 1 to 6, characterized in that, The method comprises the following steps: Turning on the multi-droplet automatic titration device to generate a plurality of droplets with a volume meeting the expected requirements on a droplet detection platform; Detecting the current positions of the plurality of droplets by using a collection module and turning on a phased array circuit board to generate a plurality of acoustic field focal points to capture the plurality of droplets; According to the preset end position of the droplets, calculating the shortest path for the plurality of droplets to move to the end position and controlling the plurality of acoustic field focal points to move the plurality of droplets according to the shortest path; After the plurality of droplets reach the target detection positions, collecting droplet related information by using the collection module and uploading the information to an upper computer, and analyzing the composition of the droplets by the upper computer according to the related information uploaded by the collection module; After the upper computer completes the analysis of the composition of the droplets, moving the plurality of droplets to a recovery area by the plurality of acoustic field focal points generated by the phased array circuit board; Turning on a second droplet pumping module of a recovery device to recover the droplets and store the droplets in a second liquid storage module.

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