Ultrasonic-based in-needle mixing method and system

Abstract

The present application relates to the technical field of in-needle mixing, and specifically to an ultrasound-based in-needle mixing method and system, comprising: providing an in-needle mixing device; obtaining a first ultrasonic characteristic and a second ultrasonic characteristic of the sample, the first ultrasonic characteristic comprising: a test mixing degree obtained by mixing a type I sample and a type II sample at multiple ultrasonic frequencies for a target duration; the second ultrasonic characteristic comprising: a test damage degree obtained by mixing a type I sample at multiple ultrasonic frequencies for a target duration; selecting a reference ultrasonic frequency corresponding to the current sample from the first ultrasonic characteristic and the second ultrasonic characteristic according to the type of item to be tested; and controlling the in-needle mixing process according to the reference ultrasonic frequency. The in-needle ultrasonic mixing technology proposed by the present invention can simplify the sample mixing operation and alleviate or avoid the degree of damage to special samples such as cells during the ultrasonic process.

Description

Technical Field

[0001] The present invention relates to the field of detection technology, and in particular to an ultrasound-based intra-needle mixing method and system. Background Art

[0002] In some biomedical testing projects, mixing of two liquid samples is often involved. Currently, when instruments mix two liquid samples, they generally draw the liquid (also called sample or reagent) into a fixed consumable container and then mix them. However, this mixing method has very low mixing efficiency and has many drawbacks, such as wasting consumables, time, samples and reagents.

[0003] In this regard, a mixing method of mixing in a needle has been proposed in the prior art.

[0004] For example, patent application CN108854650A discloses a mixing device and a mixing method thereof. Specifically, the device adopts an eccentric motion mode, so that a power mechanism can drive the eccentric mechanism and the sample dispenser to perform eccentric motion synchronously, thereby realizing the stirring function of the mixing device and promoting the mixing of the mixed liquid.

[0005] For another example, patent application CN114624090A discloses a needle mixing structure and automated detection equipment. Specifically, it causes the tail of the reagent needle to move at high frequency within a certain circumference to promote rotation and mixing of the liquid in the reagent needle. The mixing structure includes a movable mechanism that drives the reagent needle to vibrate when a power device is activated and drives the reagent needle to return to its original position when the power device is deactivated.

[0006] In other words, the applicant noted that even with the in-needle mixing technology, the mixing concept used is still the relatively traditional rotational mixing route, that is, rotating the liquid to mix. However, the applicant found that this mixing method based on rotational mixing will have the following drawbacks during application:

[0007] Mixing equipment is complex and bulky. For example, due to the need for rotational mixing, a movable mechanism is also included in the mixing equipment to allow the needle to rotate. This movable mechanism increases the mechanical complexity of the mixing equipment and easily leads to a larger device footprint. Furthermore, during frequent mixing, the movement of the needle also places higher demands on the mechanical stability of the equipment.

[0008] From another perspective, traditional in-needle mixing equipment has high configuration and maintenance costs.

[0009] Therefore, there is an urgent need for a miniaturized, low-cost and more stable in-needle mixing technology. Summary of the Invention

[0010] The object of the present invention is to provide a mixing device and an in-needle mixing method to partially solve or alleviate the above-mentioned deficiencies in the prior art, that is, to provide an in-needle mixing method and apparatus that can achieve miniaturization, low cost and high stability.

[0011] In order to solve the above-mentioned technical problems, the present invention specifically adopts the following technical solutions:

[0012] The first aspect of the present invention is to provide an ultrasound-based intra-needle mixing method, comprising the steps of:

[0013] S001. Provide an in-needle mixing device, comprising a needle tube connected to an ultrasonic vibration device and a connecting pipeline. A negative pressure device is provided in the connecting pipeline, and the negative pressure device is used to create a negative or positive pressure environment in the connecting pipeline to guide the corresponding sample into or out of the needle tube.

[0014] S002, obtaining a first ultrasonic feature and a second ultrasonic feature of the sample, wherein the sample includes: a first type of sample and a second type of sample, the first ultrasonic feature includes: a tested mixing degree obtained by mixing the first type of sample and the second type of sample at multiple ultrasonic frequencies for a target time; the second ultrasonic feature includes: a tested damage degree obtained by mixing the first type of sample at multiple ultrasonic frequencies for a target time;

[0015] S003, selecting at least one reference ultrasonic frequency corresponding to the current sample from the first ultrasonic feature and the second ultrasonic feature according to the type of the item to be tested;

[0016] S004: Inputting at least one reference ultrasonic frequency into the ultrasonic vibration device to promote mixing of the current sample under ultrasonic vibration.

[0017] In some embodiments, S003 includes:

[0018] S0031, obtaining a guiding mixing degree and a guiding breakage degree according to the type of the item to be tested;

[0019] S0032: Select at least one first ultrasonic frequency from the first ultrasonic feature according to the guided mixing degree, wherein the first ultrasonic frequency satisfies a first rule:

[0020] Test the mixing degree - guide the mixing degree to be ≥ the first threshold;

[0021] S0033: Determine whether at least one second ultrasonic frequency can be further selected from at least one first ultrasonic frequency based on the second ultrasonic feature, wherein the second ultrasonic frequency satisfies a second rule:

[0022] Test damage degree - guide damage degree ≤ second threshold;

[0023] Wherein, the first threshold value and the second threshold value are preset judgment values; if so, execute step S0034;

[0024] S0034: Determine the reference ultrasonic frequency according to at least one second ultrasonic frequency.

[0025] In some embodiments, when the result of S0033 is no, S003 further includes the steps of:

[0026] S0035, selecting at least one third ultrasonic frequency and at least one fourth ultrasonic frequency from the first ultrasonic feature and the second ultrasonic feature;

[0027] Wherein, the third ultrasonic frequency satisfies the third rule:

[0028] The second threshold value is less than the test damage degree - the guidance damage degree ≤ the third threshold value;

[0029] Test the mixing degree - guide the mixing degree to be ≥ the first threshold;

[0030] Wherein, the fourth ultrasonic frequency satisfies the second rule:

[0031] Test damage degree - guide damage degree ≤ second threshold;

[0032] The fourth threshold value is less than the test mixing degree - the guidance mixing degree and less than the first threshold value;

[0033] S0036: Determine at least one first reference ultrasonic frequency and at least one second reference ultrasonic frequency according to the at least one third ultrasonic frequency and the at least one fourth ultrasonic frequency.

[0034] In some embodiments, S004 uses a strong and weak alternating mixing method for mixing. Correspondingly, S004 includes the steps of:

[0035] S0041, using the first reference ultrasonic frequency to mix for a first time period;

[0036] S0042: Using the second reference ultrasonic frequency to mix for a second time period, wherein the second time period is greater than the first time period; and the first reference ultrasonic frequency is greater than the second reference ultrasonic frequency;

[0037] S0043, using the first reference ultrasonic frequency to mix a third duration; wherein, the first duration and the second duration are less than the corresponding target durations.

[0038] In some embodiments, S004 further includes the steps of:

[0039] S0041-S0043 were repeated at least once.

[0040] In some embodiments, the steps include:

[0041] S005, recording the actual mixing degree and actual damage degree of the current sample;

[0042] S006: Determine whether the actual mixing degree and the actual damage degree meet the requirements of the guide mixing degree and the guide damage degree; if so, execute the following steps:

[0043] S007: Record the current ultrasound working data and update the first ultrasound feature or the second ultrasound feature accordingly.

[0044] In some embodiments, the inner diameter of the needle tube 12 is 0.4-0.9 mm.

[0045] The present invention also provides an ultrasound-based intra-needle mixing system, comprising:

[0046] The needle tube comprises a first needle tube and a second needle tube, wherein the inner diameter of the first needle tube gradually decreases from the first end to the second end thereof, and the second end of the first needle tube is provided with an anti-drip portion; wherein the anti-drip portion comprises: at least two extended wall surfaces extending outwardly from the second end of the first needle tube, with a gap formed between adjacent two extended wall surfaces;

[0047] a connecting pipeline, a first end of the connecting pipeline being connected to the second needle tube;

[0048] an ultrasonic vibration device connected to the second needle tube;

[0049] A negative pressure device is provided on the connecting pipeline and is used to form a negative pressure or positive pressure environment in the connecting pipeline to guide the corresponding sample to enter or exit the needle tube.

[0050] In some embodiments, the occupied area of one or more of the vacant surfaces is 1 / 3 to 1 / 2, and the occupied area = the total area of all the vacant surfaces / (the total area of all the vacant surfaces + the total area of all the extended wall surfaces); and / or, the inner diameter of the second needle tube is 0.4-0.9 mm.

[0051] In some embodiments, the edge of the corner of the extended wall is a first arcuate edge, and a second arcuate edge is provided transitionally between adjacent extended walls; wherein the first arcuate edge, the second arcuate edge, and the first arcuate edge are connected in sequence to form the corresponding vacant surface.

[0052] Beneficial technical effects:

[0053] In some biological testing projects, biological samples need to be mixed for testing. Traditional manual mixing methods are usually cumbersome, so in-needle mixing technology has been proposed in the prior art to improve mixing efficiency and reduce sample waste during the mixing process.

[0054] The applicant noted that in order to mix the liquid sample in a narrow needle tube, traditional needle mixing requires the use of a rotating needle tube to guide the liquid sample to move significantly in the tube, thereby achieving the purpose of mixing.

[0055] However, this rotating needle requires a complex mechanism to be configured on the in-needle mixing device, which results in high cost and large size. Moreover, this complex mechanism is prone to instability after long-term operation.

[0056] In stark contrast to the traditional rotating needle method, the present invention proposes a method for ultrasonic mixing based on a fixed position (or an in-situ ultrasonic mixing method). Specifically, in order to fully mix a trace amount of sample in a fixed and limited needle, the present invention provides a frequency screening method that alternately screens ultrasonic frequencies based on three factors: sample type, mixing index, and damage index. This screening method can provide relatively reliable ultrasonic conditions for mixing multiple types of samples, and thus avoid damage to core detection objects (such as cells) and affect detection results while meeting the requirements for sufficient mixing.

[0057] It's important to note that, especially when testing patients, for example, when immune function is abnormal, the stability of their red blood cell membranes decreases, and the fragility of red blood cells increases, making them more susceptible to rupture during the mixing process. Furthermore, during the mixing process, liquid from the needle tip can easily spill and contaminate the operating platform.

[0058] In response to this special mixing requirement, the present application proposes a mixing solution with less restrictions on the operating environment and sample type, specifically adopting an in-situ ultrasonic mixing solution with an anti-drip function. Among them, the extended wall surface of the anti-drip area cooperates with the vacant surface to reduce the adhesion ability of the droplets on the head of the needle tube, causing the droplets of liquid to drip quickly at the head of the needle tube to avoid excessive liquid remaining in the head. Furthermore, when the needle tube moves, it is not easy for the head of the needle tube to spill liquid. In addition, liquid splashing can also be avoided during the ultrasonic vibration process. On the other hand, in the in-situ ultrasonic mixing method, the needle tube and the ultrasonic vibration device can be connected in a relatively fixed manner, thereby greatly simplifying the mechanical structure design and reducing the size of the mixing device.

[0059] Furthermore, the present invention adopts a multi-segment arc-shaped edge setting for the anti-drip portion on the fine structure of the needle, which not only plays the role of anti-drip, but also reduces the processing difficulty and improves the strength of the head.

[0060] Furthermore, when it involves mixing of multiple types of samples, especially mixing of cell samples and the mechanical strength of the cells is relatively low, the first and second ultrasonic characteristics are alternately selected and screened to obtain strong and weak vibration parameters (i.e., reference ultrasonic frequencies), and short-term strong vibrations and long-term weak vibrations are alternately performed based on the alternating screening parameters, thereby ensuring sufficient mixing of multiple types of samples while ensuring the integrity of the cells as much as possible. Furthermore, for the mixing scenarios involving multiple types of trace samples, especially cell samples, the present invention also provides a double-layer screening step to select ultrasonic frequencies. The double-layer screening step includes: 1) initial screening of the database, that is, for mixing scenarios involving three or more types of samples, the present invention can select ultrasonic features in a targeted manner based on the primary and secondary relationships of the samples; 2) fine screening of the reference characteristic frequencies, and the first and second ultrasonic characteristics are alternately screened for a second time based on the comprehensive mixing and damage indicators, thereby quickly screening out ultrasonic frequencies suitable for alternating ultrasonic vibrations.

[0061] The ultrasonic frequency selection results obtained based on the double-layer screening step can, on the one hand, ensure the effective mixing of multiple types of samples, and at the same time ensure that the core detection objects (such as the first cell) in the mixed samples can maintain a relatively complete morphology, avoiding damage to the detection objects during the mixing process, resulting in detection failure. BRIEF DESCRIPTION OF THE DRAWINGS

[0062] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the following is a brief introduction to the drawings required for the embodiments or the description of the prior art. In all drawings, similar elements or parts are generally identified by similar reference numerals. In the drawings, the various elements or parts are not necessarily drawn according to the actual scale. Obviously, the drawings described below are some embodiments of the present invention. For those of ordinary skill in the art, other drawings can also be obtained based on these drawings without inventive work.

[0063] Figure 1 A mixing device in an exemplary embodiment of the present invention;

[0064] Figure 2 is a schematic diagram of a partial structure of a first needle tube in an exemplary embodiment of the present invention;

[0065] Figure 3 is a side view of a first needle tube in an exemplary embodiment of the present invention;

[0066] Figure 4 is a first schematic diagram of a needle tube in an exemplary embodiment of the present invention;

[0067] Figure 5 is a second schematic diagram of a needle tube in an exemplary embodiment of the present invention;

[0068] Figure 6 1 is a schematic flow chart of the steps of an ultrasonic mixing method in an exemplary embodiment of the present invention;

[0069] Figure 7 FIG. 4 is a schematic diagram of a flow chart for selecting an ultrasonic frequency in an exemplary embodiment of the present invention.

[0070] Reference numerals: needle tube 1, first needle tube 11, anti-drip portion 111, extended wall surface 1111, vacant surface 1112, second needle tube 12, connecting pipeline 2, negative pressure device 3, Class I medium 4, Class II medium 5, ultrasonic vibration device 6, fixed tube sleeve 7. DETAILED DESCRIPTION

[0071] To make the purpose, technical solutions, and advantages of the embodiments of the present invention more clear, the technical solutions in the embodiments of the present invention will be clearly and completely described below in conjunction with the accompanying drawings in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative efforts are within the scope of protection of the present invention.

[0072] Herein, suffixes such as "module," "component," or "unit" used to represent elements are only used to facilitate description of the present invention and have no specific meaning. Therefore, "module," "component," or "unit" may be used interchangeably.

[0073] As used herein, terms such as "upper," "lower," "inner," "outer," "front," "back," "one end," and "the other end" indicate positions or locations based on those shown in the accompanying drawings. These terms are intended solely to facilitate and simplify the description of the present invention and are not intended to indicate or imply that the devices or components referred to must have, be constructed, or operate in a specific orientation. Therefore, they should not be construed as limitations on the present invention. Furthermore, the terms "first" and "second" are used for descriptive purposes only and should not be construed as indicating or implying relative importance.

[0074] As used herein, unless otherwise expressly specified or limited, the terms "installed," "provided with," and "connected" should be understood broadly. For example, "connected" may refer to a fixed connection, a detachable connection, or an integral connection; it may refer to a mechanical connection, a direct connection, an indirect connection via an intermediate medium, or internal communication between two components. Those skilled in the art will understand the specific meanings of the above terms in the present invention on a case-by-case basis.

[0075] As used herein, "and / or" includes any and all combinations of one or more of the associated listed items.

[0076] Herein, "plurality" means two or more than two, ie, it includes two, three, four, five, etc.

[0077] As used in this specification, the term "about" typically means + / - 5% of the stated value, more typically + / - 4% of the stated value, more typically + / - 3% of the stated value, more typically + / - 2% of the stated value, even more typically + / - 1% of the stated value, and even more typically + / - 0.5% of the stated value.

[0078] In this specification, certain embodiments may be disclosed in a format that is within a range. It should be understood that this description of "within a range" is merely for convenience and brevity and should not be interpreted as a rigid limitation on the disclosed range. Therefore, the description of a range should be considered to have specifically disclosed all possible subranges and individual numerical values within this range. For example, the description of a range of 1-6 should be considered to have specifically disclosed subranges such as from 1 to 3, from 1 to 4, from 1 to 5, from 2 to 4, from 2 to 6, from 3 to 6, etc., as well as individual numbers within this range, such as 1, 2, 3, 4, 5, and 6. Regardless of the breadth of the range, the above rules apply.

[0079] Applicants note that conventional rotary needle mixing technology presents the following difficulties: the mixing equipment is complex and bulky. For example, due to the need for rotational mixing, a movable mechanism is incorporated into the mixing device to allow the needle to rotate. This movable mechanism increases the mechanical complexity of the mixing device and tends to require more space. Furthermore, during frequent mixing, the movement of the needle places higher demands on the mechanical stability of the device.

[0080] In this regard, in contrast to the conventional rotating needle mixing technology, the present invention provides a fixed needle mixing technology, such as Figure 1 As shown, the second needle tube 12 is connected to the ultrasonic vibration device 6. When the ultrasonic vibration device 6 is activated, it directly changes the physical state of the object (such as the needle tube) through high-frequency vibration based on the interaction between ultrasound waves and the object, thereby promoting thorough mixing of the sample liquid in the needle tube under the high-frequency vibration. In particular, the present invention does not require an additional movable mechanism to enable the needle tube to rotate circumferentially or eccentrically, and the needle tube can be directly fixedly connected to the ultrasonic vibration device 6, thereby maintaining high stability even under high-frequency vibration.

[0081] However, during application, the applicant discovered that mixing samples within a narrow syringe while maintaining a fixed position placed extremely high demands on the ultrasonic frequency. However, the choice of ultrasonic frequency can also affect the quality of certain special samples. For example, particularly for tests involving cell samples, the high-frequency vibration required for mixing within the limited space of a fixed syringe can easily increase the risk of cell sample breakage, thereby reducing the reliability of test results.

[0082] In particular, for example, blood sample testing requires mixing red blood cells and plasma. However, when the subject is a patient, their red blood cells may be in an unhealthy state due to illness or other reasons. Therefore, they are very likely to break up under high-frequency vibration. This red blood cell breakage can seriously affect the reliability of the mixed sample test results.

[0083] Furthermore, in-needle mixing technology can easily contaminate the testing environment. During the sampling process, some liquid may remain at the tip of the needle. As the needle moves, this liquid may drip and contaminate the testing platform. Especially during the rotation or vibration of the needle, the contamination area may become too large.

[0084] In this regard, in stark contrast to the traditional rotational mixing technology route, the present invention provides an in-needle mixing method that inputs high-frequency vibration energy into the limited space inside the needle at a fixed position. This in-needle mixing method is more conducive to achieving the design requirements of miniaturization, high stability, and low application cost of the in-needle mixing equipment.

[0085] Furthermore, the present invention provides a fixed in-needle mixing solution with an anti-drip function. The anti-drip function can reduce the risk of droplets falling onto the detection operating table. In other words, the present invention actually provides a mixing solution with less restrictions on the operating environment and sample type (i.e., it is applicable to different types of cells). Figure 1-Figure 5 shown.

[0086] The present invention provides an in-needle mixing device (or mixing equipment) capable of achieving fixed mixing, comprising:

[0087] The needle tube 1 includes a first needle tube 11 and a second needle tube 12, wherein the needle tube preferably adopts a segmented design with an anti-drip function, for example, the inner diameter of the first needle tube 11 gradually decreases from the first end to the second end thereof, and the first needle tube 11 further includes: an anti-drip portion 111 provided at the second end thereof; wherein, see Figure 3 As shown, the anti-drip portion 111 includes: at least two extended walls 1111 (for example, see Figure 3 As shown, the cross-section of the first needle tube 11 is trapezoidal), and a gap 1112 is formed between two adjacent extended wall surfaces 1111; wherein the second end of the first needle tube is also called the head of the needle tube;

[0088] a connecting pipe 2, wherein a first end of the connecting pipe 2 is connected to the second needle tube 12;

[0089] an ultrasonic vibration device 6 connected to the second needle tube 12;

[0090] The negative pressure device 3 (also referred to as a power device) is provided on the connecting pipeline and is used to form a negative pressure or positive pressure environment in the connecting pipeline to guide the corresponding sample into or out of the needle tube.

[0091] For example, in some embodiments, see Figure 1 As shown, the mixing device includes a fixed sleeve 7, wherein one end of the fixed sleeve 7 is sleeved on the second needle tube and the other end is fixedly mounted on the ultrasonic vibration device. Therefore, the needle tube can remain in a relatively fixed position during the vibration process and only vibrates rapidly under the action of high-frequency ultrasound.

[0092] For example, in some embodiments, the ultrasonic vibration device and the second needle tube can be indirectly connected using a fixed tube sleeve 7. For example, the ultrasonic vibration device can be provided with a tubular connecting portion, and the fixed tube sleeve 7 can be directly fixed to the tubular connecting portion to indirectly connect the second needle tube and the ultrasonic vibration device.

[0093] Alternatively, in some embodiments, the needle tube can be directly set on the ultrasonic vibration device, and the fixing sleeve 7 is mainly used to strengthen the fixation of the connection between the needle tube and the ultrasonic vibration device.

[0094] For example, in some embodiments, the fixed sleeve may be provided with an internal thread for fixedly connecting with the second needle tube 12 by threaded engagement.

[0095] For example, in some embodiments, a micro ultrasonic vibrator may be used as the ultrasonic vibration device to reduce the size of the mixing device.

[0096] For example, in some embodiments, a channel may be provided in the middle of the ultrasonic vibration device to facilitate the passage of the second needle tube (or an extension of the second needle tube, or the connecting pipeline 2) through the channel, thereby enabling the rear end of the second needle tube to be connected to the negative pressure device to achieve the purpose of liquid suction and discharge with the assistance of the negative pressure device. For example, a tubular connecting portion is formed in the channel, and the tubular connecting portion can be sleeved on the second needle tube (or an extension of the second needle tube, or the connecting pipeline 2).

[0097] For example, in some embodiments, the negative pressure device 3 is connected to the second end of the connecting pipe 2, and a plunger is provided in the negative pressure device 3, and the plunger can reciprocate in a set direction to cause the needle tube to inhale and discharge the second type of medium 5 through the second end of the first needle tube 11 under the action of the plunger;

[0098] When the plunger moves in the first direction, the first type of medium 4 in the needle tube 1 is partially discharged from the first end of the second needle tube 12 under the action of the plunger, and the second type of medium 5 can be sucked in from the external space to complete the replacement of the first type of medium by the second type of medium; when the plunger moves in the second direction, the second type of medium 5 in the needle tube 1 is discharged from the second end of the first needle tube under the action of the plunger.

[0099] In some embodiments, the first type of medium 4 refers to the air in the syringe, and the second type of medium 5 refers to the liquid sample to be mixed (or simply referred to as liquid).

[0100] In this embodiment, the extended wall surface is combined with the vacant surface to reduce the adhesion of liquid droplets to the needle tip, thereby promoting rapid droplet shedding at the needle tip and preventing excess liquid from remaining there. Furthermore, when the needle moves, the tip is less likely to spill liquid.

[0101] In some embodiments, the occupied area of one or more of the vacant surfaces is 1 / 3 to 1 / 2, and the occupied area = the total area of all the vacant surfaces / the total area of all the vacant surfaces + the total area of all the extended wall surfaces.

[0102] In this embodiment, by limiting the occupied area, it is possible to prevent the droplets from hanging at the narrowest end of the extended wall.

[0103] In some embodiments, see Figure 3 As shown, the edge of the corner of the extended wall 1111 is a first arcuate edge l1, and a second arcuate edge l2 is transitionally arranged between adjacent extended walls 1111; wherein the first arcuate edge, the second arcuate edge, and the first arcuate edge are connected in sequence to form the corresponding vacant surface.

[0104] In some embodiments, the curvature radius of the first curved edge l1 is smaller than the curvature radius l2 of the second curved edge. In other words, the smoothness of the first curved edge is higher than the smoothness of the second curved edge.

[0105] Preferably, see Figure 3 As shown, the gap is formed by connecting two adjacent extended wall surfaces with multiple curved edges. The curved edges of the head section round off the corners of the protruding extended wall surface. This rounding provides some protection for the anti-drip portion, preventing damage to the head due to improper contact during use. Furthermore, the slightly larger radius of curvature of the second curved edge can also reduce the machining difficulty of small structures, while also preventing excessive stress concentration in localized areas and ensuring the strength of the small structure.

[0106] It's worth noting that sampling needles are typically quite thin, especially where the first needle tapers toward its second end. Therefore, machining this tiny structure is extremely challenging. The multi-segmented curved edge design in this embodiment not only prevents dripping but also reduces machining complexity and enhances the strength of the tip.

[0107] In addition, it is worth noting that in this embodiment, reciprocating motion is used to mix the liquid so that the needle tip of the needle tube can basically maintain its position unchanged during the mixing process, thereby improving the structural stability of the needle tube during the mixing process and avoiding splashing of the liquid.

[0108] Moreover, this fixed needle type (i.e., no circular motion or eccentric motion is required) can further simplify the mechanical design difficulty of the mixing equipment.

[0109] In some embodiments, the negative pressure device is a plunger pump.

[0110] In some embodiments, the inner diameter of the second needle tube 12 is 0.4-0.9 mm.

[0111] In some embodiments, the second syringe is preferably suitable for mixing 10-20 μL of the second type of medium (or sample).

[0112] In some embodiments, a hydrophobic coating is provided on the inner wall of the needle tube.

[0113] Preferably, in this embodiment, the inner wall surface of the needle tube has a hydrophobic property, thereby limiting the spreading ability of the liquid on the inner wall, thereby reducing the contact area between the liquid and the surface, and further hindering the generation of bubbles.

[0114] In some embodiments, the inner wall of the needle tube is water-polished.

[0115] Preferably, the inner wall of the needle tube in this embodiment adopts a smooth surface (that is, the smoothness can be improved by water grinding). This can prevent the generation of bubbles on the one hand, and on the other hand, prevent the red blood cells from being broken by external stimuli during the reciprocating movement.

[0116] In some embodiments, the needle is a glass needle.

[0117] In some embodiments, the needle tube is a steel needle.

[0118] As previously mentioned, the present invention provides a fixed in-needle mixing method (or, in-situ ultrasonic mixing method) that utilizes relatively high-frequency ultrasound to mix the sample within the confined space within the needle. To balance mixing uniformity with the risk of sample damage, the present invention provides an ultrasonic mixing method that selects ultrasonic frequencies based on key characteristics.

[0119] The present invention also provides an ultrasound-based needle mixing method, see Figure 6 As shown, the steps include:

[0120] S001. Provide an in-needle mixing device, comprising a needle tube connected to an ultrasonic vibration device and a connecting pipeline. A negative pressure device is provided in the connecting pipeline, and the negative pressure device is used to create a negative or positive pressure environment in the connecting pipeline to guide the corresponding sample into or out of the needle tube.

[0121] The in-needle mixing device may be the in-needle mixing device described in any one of the embodiments of the present invention.

[0122] S002, obtaining a first ultrasonic feature and a second ultrasonic feature of the sample, wherein the sample includes: a first type of sample and a second type of sample, the first ultrasonic feature includes: a tested mixing degree obtained by mixing the first type of sample and the second type of sample at multiple ultrasonic frequencies for a target time; the second ultrasonic feature includes: a tested damage degree obtained by mixing the first type of sample at multiple ultrasonic frequencies for a target time;

[0123] Preferably, one type of sample is usually a liquid sample containing cells. Further, one type of sample is usually a liquid sample containing cells to be tested (such as red blood cells, white blood cells, etc.), and the cells to be tested can be the test subjects of the items to be detected.

[0124] For example, in some embodiments, before conducting formal ultrasonic vibration mixing, multiple groups of preliminary experiments can be carried out to obtain multiple data records. A data record may include: vibration frequency, vibration time (equivalent to the target duration), test results - test mixing degree and / or test damage degree. Based on multiple data records, a feature database such as the corresponding first ultrasonic feature and the second ultrasonic feature can be statistically generated. Of course, in other embodiments, the above-mentioned data records can also be application records generated in real time during the application process (such as when the operator completes a real detection operation, that is, the ultrasonic parameters selected for the process and the final test results are synchronously recorded), that is, the feature database can also be gradually expanded as actual detection data is generated.

[0125] For example, in some embodiments, the feature database may record test results of different ultrasonic frequencies and different vibration durations.

[0126] For example, in some embodiments, the first ultrasonic characteristic may be a first fitting curve formed based on multiple data records, where each data record includes vibration time, vibration frequency, and a tested mixing result (e.g., mixing degree). Correspondingly, a curve representing the relationship between vibration frequency and mixing degree can be fitted based on the multiple data records. Using a rectangular coordinate system as an example, the vibration frequency can be represented by the X-axis, and the mixing degree can be represented by the Y-axis.

[0127] Similarly, the second super-feature can also be a second fitted curve formed based on multiple data records. A data record includes vibration time, vibration frequency, and the mixing result of the test (e.g., degree of damage). Correspondingly, a curve representing the relationship between vibration frequency and degree of damage can be fitted based on the multiple data records. Using a rectangular coordinate system as an example, the X-axis represents vibration frequency, and the Y-axis represents degree of damage.

[0128] S003, selecting at least one reference ultrasonic frequency corresponding to the current sample from the first ultrasonic feature and the second ultrasonic feature according to the type of the item to be tested;

[0129] Preferably, the samples to be mixed include: sample A and sample B, wherein sample A contains cells. Correspondingly, the first ultrasonic feature is selected or collected by sample A and sample B (equivalent to Class I sample and Class II sample), that is, the first ultrasonic feature is preferentially the mixing parameter of the actual samples A and sample B to be mixed, and the second ultrasonic feature is selected or collected by sample A (equivalent to Class II sample).

[0130] S004: Inputting at least one reference ultrasonic frequency into the ultrasonic vibration device to promote mixing of the current sample under ultrasonic vibration.

[0131] For example, in some embodiments, see Figure 7 As shown, S003 includes:

[0132] S0031, obtaining a guiding mixing degree and a guiding breakage degree according to the type of the item to be tested;

[0133] For example, in some embodiments, the types of items to be tested are divided into quantitative testing and qualitative testing, and different item types may have different requirements for mixing quality (such as mixing degree, damage degree).

[0134] For example, in some embodiments, the degree of mixing of a liquid sample can be assessed by its appearance (e.g., color or turbidity). Alternatively, in some embodiments, the degree of mixing of a liquid sample can be assessed by quantitative testing methods such as chromatography, mass spectrometry, and infrared spectroscopy. Specifically, scores can be set for different degrees of mixing, and the degree of mixing of a sample can be scored based on the above appearance observations or quantitative tests.

[0135] For example, in some embodiments, the degree of cell damage in a cell sample can be characterized by the damage rate as follows: Damage rate = (number of damaged cells / total number of cells) × 100%. For example, in some embodiments, direct counting methods (microscope observation, such as trypan blue staining or fluorescent staining) can be used for statistical analysis. Alternatively, optical density measurement (an indirect method) can be used for statistical analysis.

[0136] S0032: Select at least one first ultrasonic frequency from the first ultrasonic feature according to the guided mixing degree, wherein the first ultrasonic frequency satisfies a first rule:

[0137] ;

[0138] Preferably, in some embodiments, the first rule is: ; That is, at least one ultrasonic frequency that can be fully mixed is selected from the first ultrasonic feature.

[0139] For example, when the first ultrasonic feature is a first fitting curve obtained by fitting multiple data records, a first line segment where Y ≥ the degree of mixing can be selected from the first fitting curve, and at least two ultrasonic frequencies corresponding to the first line segment can be recorded. For example, the ultrasonic frequencies corresponding to the two endpoints of the line segment can be obtained.

[0140] S0033: Determine whether at least one second ultrasonic frequency can be further selected from at least one first ultrasonic frequency based on the second ultrasonic feature, wherein the second ultrasonic frequency satisfies a second rule:

[0141] ;

[0142] The first threshold and the second threshold are preset judgment values; preferably, the first threshold and the second threshold can be preset by the user.

[0143] For example, in some embodiments, a suitable second ultrasonic frequency can be further selected from the first line segment. Specifically, the test damage degree corresponding to at least two ultrasonic frequencies in the first line segment can be queried respectively, and the one that meets the following rules can be selected: The ultrasonic frequency is used as the second ultrasonic frequency.

[0144] If yes, execute step S0034;

[0145] S0034: Determine the reference ultrasonic frequency according to at least one second ultrasonic frequency.

[0146] For example, in some embodiments, a second ultrasonic frequency may be selected from a plurality of second ultrasonic frequencies as a reference ultrasonic frequency.

[0147] For another example, in some embodiments, at least two of the multiple second ultrasonic frequencies may be selected as reference ultrasonic frequencies, and the vibration process may use the multiple reference ultrasonic frequencies for alternating vibration.

[0148] For another example, in some embodiments, an average of multiple second ultrasonic frequencies may be calculated as the reference ultrasonic frequency.

[0149] It should be noted that for different types of testing items, the types and quantities of samples required for mixing are different, so the mixing indicators and damage requirements involved are also quite different.

[0150] Especially when facing the mixing of multiple types of samples, and the samples involve special types of cells, such as the red blood cells of anemic patients, which are relatively fragile and easily damaged during the ultrasound process. Therefore, when it comes to the mixing of red blood cells or other cells with low mechanical strength, it will be difficult to take into account the requirements of mixing index and damage index. In this regard, the present invention provides a method for alternately screening ultrasonic frequencies based on sample type, mixing index, and damage index for the in-situ mixing method to solve the mixing needs of cells with low mechanical strength.

[0151] Furthermore, in some embodiments, when the result of S0033 is no, S003 further includes the steps of:

[0152] S0035, selecting at least one third ultrasonic frequency and at least one fourth ultrasonic frequency from the first ultrasonic feature and the second ultrasonic feature;

[0153] Wherein, the third ultrasonic frequency satisfies the third rule:

[0154] ;

[0155] ;

[0156] Wherein, the fourth ultrasonic frequency satisfies the second rule:

[0157] ;

[0158] ;

[0159] S0036: Determine at least one first reference ultrasonic frequency and at least one second reference ultrasonic frequency according to the at least one third ultrasonic frequency and the at least one fourth ultrasonic frequency.

[0160] For example, a first reference ultrasonic frequency may be selected from at least one third ultrasonic frequency, or an average value of at least one third ultrasonic frequency may be calculated as the first reference ultrasonic frequency.

[0161] For example, a second reference ultrasonic frequency may be selected from at least one fourth ultrasonic frequency, or an average value of at least one fourth ultrasonic frequency may be calculated as the second reference ultrasonic frequency.

[0162] It is understandable that one or more of the above thresholds can be preset by the user.

[0163] In this embodiment, at the selected third ultrasonic frequency, the sample mixing degree is relatively high, but the risk of cell damage may also be high. Therefore, the vibration at the third ultrasonic frequency can also be called strong vibration. At the selected fourth ultrasonic frequency, the sample mixing degree is relatively low, but the risk of cell damage is also reduced. Therefore, the vibration at the fourth ultrasonic frequency can also be called weak vibration.

[0164] In some embodiments, S004 uses a strong and weak alternating mixing method for mixing. Correspondingly, S004 includes the steps of:

[0165] S0041, using the first reference ultrasonic frequency to mix for a first time period;

[0166] S0042, using the second reference ultrasonic frequency to mix for a second duration, wherein the second duration is greater than the first duration, and the first reference ultrasonic frequency is greater than the second reference ultrasonic frequency;

[0167] S0043, using the first reference ultrasonic frequency to mix a third duration; wherein, the first duration and the second duration are less than the corresponding target durations.

[0168] Preferably, the second duration is greater than the first duration and greater than the third duration.

[0169] In this embodiment, alternating strong and weak vibrations are performed based on the strong and weak vibration data obtained by the above-mentioned alternating screening, thereby ensuring sufficient mixing of multiple types of samples while ensuring the integrity of the cells as much as possible.

[0170] In some embodiments, S004 further includes the steps of:

[0171] S0041-S0043 were repeated at least once.

[0172] It is noteworthy that the alternating vibration scheme based on alternating screening proposed in the present invention can further limit the degree of damage to the cell sample and avoid damage to the sample on the basis of ensuring uniform mixing of the sample.

[0173] Furthermore, for a scenario where multiple types of samples are mixed, the samples may contain multiple types of cells, such as at least two samples are cells or liquid samples containing at least one type of cells. Step S002 includes:

[0174] (1) Determine the first cell and the second cell according to the type of the test item, wherein the first cell can directly or indirectly reflect the required test index; conversely, the second cell can be a cell incidental to the liquid sample, which under normal conditions generally does not cause adverse effects or interference on the final test result.

[0175] It is understood that the test items referred to in the present invention may be conventional medical tests or biological test items, and thus the selection of the first cell and the second cell may be specified according to the test item, or the test item may pre-record the type information of the first cell and the second cell;

[0176] (2) Select at least one main sample from at least two samples, where the main sample is a sample with a sample content greater than a preset proportion;

[0177] (3) selecting a corresponding first ultrasonic feature according to at least one main sample;

[0178] For example, in some embodiments, the selected first ultrasound feature is a first fitting curve.

[0179] Preferably, in some embodiments, two main samples may be selected, and the first ultrasonic features corresponding to the first type of sample and the second sample corresponding to the two main samples may be selected.

[0180] Preferably, in some embodiments, the two main samples are of exactly the same type as the first category sample and the second category sample.

[0181] Alternatively, in some embodiments, the first or second type of samples referred to may be samples having sample properties (such as liquid viscosity, density, particle size, such as cell particle size) that are the same or similar to those of the main sample. For example, similar means that the degree of difference between the sample properties of the two is within a preset difference ratio, such as within 10% or even within 20%.

[0182] (4) selecting a corresponding second ultrasonic feature according to the first cell;

[0183] Typically, the first cell is a cell contained in the selected first or second type of sample, or the first cell is the first or second type of sample.

[0184] For example, in some embodiments, the actual process in step (4) is: selecting the second ultrasonic feature corresponding to the first type of sample or the second type of sample containing the first cell.

[0185] For example, in some embodiments, a second fitting curve corresponding to the first cell (or a sample containing the first cell) may be selected.

[0186] In this embodiment, for the mixing scenario of multiple types of samples, a method for screening ultrasonic frequencies based on a main sample is proposed.

[0187] In some embodiments, mixing of two or more cell types is involved, such as mixing of cells A, B, and C. However, if cell C is the primary target for detection, and cell C and cell A are the two cell types with the highest content, then cells C (or, in other words, a liquid containing cell C) and cells A (or, in other words, a liquid containing cell A) can be used as primary samples, and a first ultrasonic feature can be selected based on cells C and A (for example, in the early stages of application, a pre-mixing test can be performed on cells C and A, and the corresponding mixing results, such as a first fitting curve, can be recorded). Furthermore, a second ultrasonic feature of cell C can be obtained (this data can also be obtained through a pre-mixing test). Finally, a reference ultrasonic feature can be obtained by comprehensively selecting the first and second ultrasonic features.

[0188] Notably, for mixing scenarios involving multiple sample types, the present invention provides a two-tiered screening process for selecting ultrasonic frequencies. This two-tiered screening process includes: 1) initial database screening. Specifically, for mixing scenarios involving three or more sample types, the present invention selects ultrasonic features based on the primary and secondary relationships of the samples; and 2) fine-scale screening with reference to characteristic frequencies. This process alternately screens the first and second ultrasonic features based on mixing and damage indicators, thereby rapidly selecting an ultrasonic frequency suitable for alternating ultrasonic vibrations.

[0189] The ultrasonic frequency selection results obtained based on the double-layer screening step can, on the one hand, ensure the effective mixing of multiple types of samples, and at the same time ensure that the core detection objects (such as the first cell) in the mixed samples can maintain a relatively complete morphology, avoiding damage to the detection objects during the mixing process, resulting in detection failure.

[0190] In some embodiments, the steps include:

[0191] S005, recording the actual mixing degree and actual damage degree of the current sample;

[0192] S006: Determine whether the actual mixing degree and the actual damage degree meet the guide mixing degree and the guide damage degree requirements (for example, the actual mixing degree and the actual damage degree meet the first and second rules respectively). If so, execute the following steps:

[0193] S007: Record the current ultrasound working data and update the first ultrasound feature or the second ultrasound feature accordingly.

[0194] In some embodiments, the inner diameter of the needle tube 12 is 0.4-0.9 mm.

[0195] The mixing method is described below by taking the mixing process of samples such as plasma and red blood cells as an example:

[0196] First, a sampling container can be set on the equipment operating table, which carries the blood sample after centrifugation, and the sample consists of a plasma layer, a platelet and white blood cell layer, and a red blood cell layer from top to bottom.

[0197] Then, a suitable amount of red blood cells (which in some embodiments corresponds to the second sample) may be drawn using a needle.

[0198] Then, the needle is raised to draw an appropriate amount of plasma (which, in some embodiments, corresponds to the first sample).

[0199] Raise the needle tube so that the lower liquid level of the mixed sample initially formed by red blood cells and plasma rises or remains above a first height H1, wherein the first height H1 is higher than or located at the dividing line between the first needle tube and the second needle tube; ensure that no droplets are hanging on the anti-drip portion 111;

[0200] For example, in some embodiments, an operator can visually observe that no liquid droplets are hanging from the anti-drip portion 111 .

[0201] Preferably, to prevent liquid splashing, the upper and lower liquid levels can be maintained between the second height H2 and the third height H3. For example, the second height H2 is preferably located above the dividing line between the first and second needle tubes. Of course, in some embodiments, the specific positions of the second and third heights can be adaptively set by the operator based on different liquid contents.

[0202] To meet the in-needle mixing requirements for a large number of sample types, the present invention provides an in-situ mixing solution that performs intensity-interactive high-frequency vibration on the samples based on partial pre-test data. This in-situ mixing solution can simplify the mechanical design difficulty of the mixing equipment and reduce application costs.

[0203] Understandably, in actual testing applications, different testing projects have different requirements for sample type, content, and even mixing level. Furthermore, different types of samples, such as different cell types, or cell samples collected from subjects with different health conditions, can often have significantly different mechanical strengths.

[0204] In this regard, the alternating ultrasonic frequency screening technology provided by the present invention based on partial pre-test data can reduce the difficulty of selecting ultrasonic conditions and reduce the workload of pre-ultrasound experiments.

[0205] It should be noted that, in this document, the terms "comprises," "includes," or any other variations thereof are intended to encompass non-exclusive inclusion, such that a process, method, article, or apparatus comprising a series of elements includes not only those elements but also other elements not explicitly listed, or elements inherent to such process, method, article, or apparatus. In the absence of further limitations, an element defined by the phrase "comprising a ..." does not exclude the presence of other identical elements in the process, method, article, or apparatus comprising the element.

[0206] Through the description of the above embodiments, those skilled in the art can clearly understand that the above-mentioned embodiment methods can be implemented by means of software plus the necessary general hardware platform, and of course can also be implemented by hardware, but in many cases the former is a better embodiment. Based on this understanding, the technical solution of the present invention is essentially or the part that contributes to the prior art can be embodied in the form of a software product, which is stored in a storage medium (such as ROM / RAM, magnetic disk, optical disk), including a number of instructions for enabling a computer terminal (which can be a mobile phone, computer, server, or network device, etc.) to execute the methods described in each embodiment of the present invention.

[0207] The embodiments of the present invention are described above in conjunction with the accompanying drawings, but the present invention is not limited to the above-mentioned specific implementation methods. The above-mentioned specific implementation methods are merely illustrative and not restrictive. Under the guidance of the present invention, ordinary technicians in this field can also make many forms without departing from the scope of protection of the present invention and the claims, all of which are protected by the present invention.

Claims

1. An ultrasound-based in-needle mixing method, characterized in that: Including steps: S001. Provide an in-needle mixing device, comprising a needle tube connected to an ultrasonic vibration device and a connecting pipeline. A negative pressure device is provided in the connecting pipeline, and the negative pressure device is used to create a negative or positive pressure environment in the connecting pipeline to guide the corresponding sample into or out of the needle tube. S002, obtaining a first ultrasonic feature and a second ultrasonic feature of the sample, wherein the sample includes: a first type of sample and a second type of sample, the first ultrasonic feature includes: a tested mixing degree obtained by mixing the first type of sample and the second type of sample at multiple ultrasonic frequencies for a target time; the second ultrasonic feature includes: a tested damage degree obtained by mixing the first type of sample at multiple ultrasonic frequencies for a target time; S003, selecting at least one reference ultrasonic frequency corresponding to the current sample from the first ultrasonic feature and the second ultrasonic feature according to the type of the item to be tested; wherein S003 includes: S0031, obtaining a guiding mixing degree and a guiding breakage degree according to the type of the item to be tested; S0032: Select at least one first ultrasonic frequency from the first ultrasonic feature according to the guided mixing degree, wherein the first ultrasonic frequency satisfies a first rule: Test the mixing degree - guide the mixing degree to be ≥ the first threshold; S0033: Determine whether at least one second ultrasonic frequency can be further selected from at least one first ultrasonic frequency based on the second ultrasonic feature, wherein the second ultrasonic frequency satisfies a second rule: Test damage degree - guide damage degree ≤ second threshold; Wherein, the first threshold value and the second threshold value are preset judgment values; if so, execute step S0034; S0034, determining the reference ultrasonic frequency according to at least one second ultrasonic frequency; S004: Inputting at least one of the reference ultrasonic frequencies into the ultrasonic vibration device to promote mixing of the current sample under ultrasonic vibration.

2. The ultrasound-based in-needle mixing method according to claim 1, characterized in that: When the result of S0033 is no, S003 further includes the steps of: S0035, selecting at least one third ultrasonic frequency and at least one fourth ultrasonic frequency from the first ultrasonic feature and the second ultrasonic feature; Wherein, the third ultrasonic frequency satisfies the third rule: ； ； Wherein, the fourth ultrasonic frequency satisfies the second rule: ； ； S0036: Determine at least one first reference ultrasonic frequency and at least one second reference ultrasonic frequency according to the at least one third ultrasonic frequency and the at least one fourth ultrasonic frequency.

3. The ultrasound-based in-needle mixing method according to claim 2, characterized in that: In S004, a strong and weak alternating mixing method is used for mixing. Correspondingly, S004 includes the steps of: S0041, mixing for a first time using the first reference ultrasonic frequency; S0042: Using the second reference ultrasonic frequency to mix for a second duration, wherein the second duration is greater than the first duration, and the first reference ultrasonic frequency is greater than the second reference ultrasonic frequency; S0043, using the first reference ultrasonic frequency to mix a third duration; wherein, the first duration and the second duration are less than the corresponding target durations.

4. The ultrasound-based in-needle mixing method according to claim 3, characterized in that: S004 also includes the steps of: S0041-S0043 were repeated at least once.

5. The ultrasound-based in-needle mixing method according to claim 4, characterized in that: Including steps: S005, recording the actual mixing degree and actual damage degree of the current sample; S006: Determine whether the actual mixing degree and the actual damage degree meet the requirements of the guide mixing degree and the guide damage degree; if so, execute the following steps: S007: Record the current ultrasound working data and update the first ultrasound feature or the second ultrasound feature accordingly.

6. The ultrasound-based in-needle mixing method according to any one of claims 1 to 5, characterized in that: The inner diameter of the needle tube is 0.4-0.9 mm.

7. An ultrasound-based in-needle mixing system, characterized in that: include: A needle tube (1), comprising a first needle tube (11) and a second needle tube (12), wherein the inner diameter of the first needle tube (11) gradually decreases in a direction from the first end to the second end thereof, and the second end of the first needle tube (11) is provided with an anti-drip portion (111); wherein the anti-drip portion (111) comprises: at least two extended wall surfaces (1111) formed by extending outward from the second end of the first needle tube (11), and a vacant surface (1112) is formed between two adjacent extended wall surfaces (1111); A connecting pipeline (2), wherein a first end of the connecting pipeline (2) is connected to the second needle tube (12); an ultrasonic vibration device (6), the ultrasonic vibration device (6) being connected to the second needle tube (12); A negative pressure device is provided on the connecting pipeline and is used to form a negative pressure or positive pressure environment in the connecting pipeline to guide the corresponding sample to enter or exit the needle tube.

8. The in-needle mixing system according to claim 7, characterized in that: The occupied area of one or more of the vacant surfaces is 1 / 3 to 1 / 2, and the occupied area = the total area of all the vacant surfaces / (the total area of all the vacant surfaces + the total area of all the extended wall surfaces); and / or the inner diameter of the second needle tube (12) is 0.4-0.9 mm.

9. The in-needle mixing system according to claim 8, characterized in that: The edges of the corners of the extended wall surfaces (1111) are first arcuate edges, and second arcuate edges are transitionally provided between adjacent extended wall surfaces (1111); wherein the first arcuate edge, the second arcuate edge, and the first arcuate edge are sequentially connected to form the corresponding vacant surface.

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