CN111893036A

CN111893036A - Bacterium counting assembly with bubble mixing mechanism

Info

Publication number: CN111893036A
Application number: CN201911276803.9A
Authority: CN
Inventors: 崔璟; 唐明忠; 许朋
Original assignee: Beijing Xinji Jinnuo Medical Devices Co ltd; Shandong Xinke Biological Technology Co ltd
Current assignee: BEIJING XINJI JINNUO MEDICAL EQUIPMENT Co.,Ltd.
Priority date: 2019-12-12
Filing date: 2019-12-12
Publication date: 2020-11-06

Abstract

The invention provides a bacteria counting device, comprising: the sampling assembly comprises a three-dimensional motion mechanical arm and a sampling needle, the three-dimensional motion mechanical arm is used for driving the sampling needle to suck a bacterial sample to be counted from the reagent plate, and the sampling needle is used for adding the sucked bacterial sample to be counted into the counting cell assembly; the pump is connected with the sampling needle through a liquid path and is used for uniformly mixing the bacteria sample to be counted, which is sucked by the sampling needle, wherein when the pump works in a first mode, the liquid in the liquid path is pushed to flow to the sampling needle, and when the pump works in a second mode, the liquid in the sampling needle is sucked into the liquid path. Inhale before the appearance with inhale twice mixing behind the appearance, guarantee the sample intensive mixing that awaits measuring, guarantee that the statistics of bacterium number is more accurate in the testing process, the mixing operation is inside equipment, does not need other supplementary mixing equipment firstly, but the pollution rate is reduced.

Description

Bacterium counting assembly with bubble mixing mechanism

Technical Field

The invention relates to the field of biology, and further relates to a bacteria counting device, in particular to a bacteria counting device with a bubble uniformly mixing mechanism.

Background

A cytometer is generally an instrument for measuring the number of platelets, white blood cells, red blood cells, and the like. The full-automatic cell counter is widely applied, and the technical scheme of the coulter principle analysis method is an internationally recognized standard control method for measuring the sizes of cells and particles and always occupies an important position in hematology analysis.

The resistance counting method, i.e. the coulter method, measures the number of particles using the principle of small-hole resistance. The particles follow the liquid flow during the measurement. When it passes through the small hole, the cross-sectional area of the small hole is reduced, the resistance between the two electrodes is increased, the voltage is raised, and a voltage pulse signal is generated. The number of particles can be counted by the instrument as long as each pulse is accurately measured, and the instrument is also a common design basis for most blood analyzers.

The present design is applied to counting of bacteria, and the following problems still exist: the prior art has various liquid mixing modes, such as oscillation, ultrasound and the like; but these devices are all ancillary single functions of other devices. Secondly, the existing equipment is used for uniformly mixing bacteria, and the pollution can be caused by improper operation.

Therefore, a bacteria counting device with a bubble mixing mechanism is to be provided.

Disclosure of Invention

The embodiment of the invention provides a bacteria counting device, which at least solves the technical problem that the prior art causes pollution due to improper operation of a micro-particle counting device in a bacteria mixing process.

The embodiment of the invention provides a bacteria counting device, which comprises: the sampling assembly comprises a three-dimensional motion mechanical arm and a sampling needle, the three-dimensional motion mechanical arm is used for driving the sampling needle to suck a bacterial sample to be counted from the reagent plate, and the sampling needle is used for adding the sucked bacterial sample to be counted into the counting cell assembly; and the pump is connected with the sampling needle through a liquid path and is used for uniformly mixing the bacteria sample to be counted, which is sucked by the sampling needle, wherein when the pump works in a first mode, the pump pushes the liquid in the liquid path to flow to the sampling needle, and when the pump works in a second mode, the pump sucks the liquid in the sampling needle into the liquid path.

Optionally, the sampling needle is a plurality of sampling needles, the pump is connected to each sampling needle through a plurality of liquid lines, and the pump is a plunger pump or a gas pump.

Optionally, the apparatus further comprises: the plate-type plate-making machine comprises a machine frame, wherein a plate groove is formed in the machine frame; the reagent plate is clamped in the plate groove; wherein, the pump is arranged inside the frame.

Optionally, the pump is the plunger pump, wherein the plunger pump includes: a cylinder body; and a plunger which reciprocates up and down in the cylinder, wherein when the plunger moves upward, an internal volume of the cylinder is reduced to push the liquid in the liquid path to the sampling needle, and when the plunger moves downward, the internal volume of the cylinder is increased to suck the liquid in the sampling needle into the liquid path.

Optionally, the plunger is further configured to move up and down in the cylinder before the sampling needle contacts the bacteria sample to be counted in the reagent plate but does not suck the bacteria sample to be counted, so as to mix the bacteria sample to be counted in the reagent plate.

Optionally, the apparatus further comprises: and the counting cell assembly is used for counting bacteria of the bacteria sample to be counted.

The technical scheme of the invention also provides a bacteria sampling mode, which comprises the following steps: driving a sampling needle to suck a bacterial sample to be counted from a reagent plate through a three-dimensional motion mechanical arm; uniformly mixing the bacteria sample to be counted in the sampling needle by a pump, wherein the pump is connected with the sampling needle through a liquid route, when the pump works in a first mode, the liquid in the liquid route is pushed to flow to the sampling needle, and when the pump works in a second mode, the liquid in the sampling needle is sucked into the liquid route; the sampling needle is driven to move to the counting cell assembly through the three-dimensional motion mechanical arm; and controlling the sampling needle to add the sucked bacterial sample to be counted into the counting cell assembly.

Optionally, the pump comprises: cylinder body and plunger, wherein, the aforesaid of passing through in the pump to the above-mentioned sampling needle waits to count the bacterium sample and carries out the mixing, includes: and controlling the plunger to reciprocate up and down in the cylinder, wherein when the plunger moves up, the internal volume of the cylinder is reduced to push the liquid in the liquid path to flow to the sampling needle, and when the plunger moves down, the internal volume of the cylinder is increased to suck the liquid in the sampling needle into the liquid path.

Optionally, before the sampling needle is driven by the three-dimensional motion mechanical arm to suck the bacterial sample to be counted from the reagent plate, the method further includes: and (c) uniformly mixing the bacteria sample to be counted in the reagent plate by the pump before the sampling needle contacts the bacteria sample to be counted in the reagent plate but does not suck the bacteria sample to be counted.

Optionally, before the sampling needle is driven by the three-dimensional motion mechanical arm to suck the bacterial sample to be counted from the reagent plate, the method further includes: and controlling the plunger to move upwards and downwards in the cylinder body so as to uniformly mix the bacteria sample to be counted in the reagent plate.

The embodiment of the invention provides a bacteria counting method, which comprises the following steps: adding a sample of bacteria to be enumerated to a counting cell assembly, wherein said counting cell assembly comprises: the device comprises a gem hole, a front pool, a rear pool and electrodes, wherein the front pool and the rear pool are communicated through the gem hole, the liquid pressure between the front pool and the rear pool is negative pressure, the negative pressure is used for enabling a bacterial sample to be counted to enter the rear pool from the front pool through the gem hole, the electrodes are arranged on two sides of the gem hole respectively, and a preset resistance is arranged between the two sides of the gem hole under the condition that the electrodes are electrified;

detecting whether pulse signals generated by the resistance change between the two sides of the gem hole exist on the two sides of the gem hole or not, wherein the pulse signals are used for indicating that bacteria in the bacteria sample to be counted pass through the gem hole;

and under the condition that pulse signals generated on two sides of the gem hole are detected, acquiring the number of bacteria in the bacteria sample to be counted, which is determined according to the pulse signals.

Optionally, the obtaining the number of bacteria in the bacteria sample to be counted determined according to the pulse signal includes:

transmitting the pulse signal to a processing device, and acquiring the number of bacteria in the bacteria sample to be counted, which is sent by the processing device, wherein the number of bacteria in the bacteria sample to be counted is determined according to the bacteria characteristic data represented by the pulse signal; or

And determining the number of bacteria in the bacteria sample to be counted according to the bacteria characteristic data represented by the pulse signal.

Optionally, the diameter of the gem hole is a diameter within a first target diameter range, wherein the first target diameter range is used for allowing only one bacterium to pass through the gem hole at a time when the bacteria in the bacteria sample to be counted pass through the gem hole; or

The diameter of the gem hole is within a second target diameter range, wherein the second target diameter range is used for allowing a plurality of bacteria to pass through the gem hole at a time when the bacteria in the bacteria sample to be counted pass through the gem hole.

One of the electrodes is disposed on each side of the gem hole, and the bacteria are non-conductive, so that the bacteria generate voltage pulse signals when passing through the gem hole, and the number of bacteria in the bacteria sample to be counted can be determined according to the pulse signals, and the pulse signals can be selected as voltage pulse signals.

The bacteria characteristic data represented by the pulse signal comprises: amplifying and gaining the pulse signal through a conditioning circuit, filtering noise through low-pass filtering, and filtering out an overrun amplitude value through buffer amplitude limiting; and identifying the signal with the bacteria characteristic data in the pulse signal through algorithms such as pulse identification, slope identification, peak detection, trough detection, broadband detection and the like.

Blocking phenomenon (hole blocking) appears in above-mentioned precious stone hole, divide into totally block up the hole and incompletely block up the hole, and complete blocking phenomenon appears in above-mentioned precious stone hole promptly and incomplete blocking phenomenon appears in above-mentioned precious stone hole.

If the hole is completely blocked, the counting amount is abnormal and a correct result cannot be counted, and the recoiling assembly or the burning assembly is adopted to eliminate the hole blocking phenomenon; but if the incomplete hole blocking happens, the data can be displayed, the test result is directly influenced, whether the incomplete hole blocking phenomenon occurs can be distinguished from the observation counting time, namely, the observation counting time has a reference value, if the bacteria counting device works normally, the micropores are unobstructed, the time for sucking the bacteria sample to be counted is fixed, when the counting time is prolonged, the incomplete hole blocking phenomenon occurs on a detector of the bacteria counting device is shown, optionally, an algorithm is adopted to judge the number exceeding the limit when the hole blocking occurs on the other scheme, so that the data is judged to be inaccurate, and the hole blocking phenomenon is judged or the external interference is received.

Because under the device normal operating condition, the count time fixed value has been established, the count time is even, because under the normal operating condition, aperture voltage is stable in certain extent basically, if take place aperture voltage rising or count quantity unusual condition, then prove stifled hole or impurity interference phenomenon appear in above-mentioned precious stone hole, stifled hole reason has a lot of, most of cases are because multiple bacterium mixes inhomogeneous, perhaps wash the precious stone hole often, then the piling up of non-counting material can appear to produce stifled hole.

Optionally, a mode of judging whether the hole is blocked or not through a voltage interval is provided, that is, the voltage is divided into 3 grades, namely, the voltage is normal, higher or abnormal, when the voltage is higher, the hole blocking phenomenon is generated by a detector of the bacteria counting device, the higher is a micro hole blocking phenomenon (namely, an incomplete hole blocking phenomenon), the abnormal is a complete hole blocking phenomenon, and the normal is a non-hole blocking state; if the voltage of the small hole rises or the counting amount is abnormal or the base line is judged to be abnormal, the phenomenon that the gem hole is blocked or impurities or interference occur is proved.

Under normal conditions: the middle liquid port of the rear tank is under negative pressure, the rear tank is provided with three channels, the upper channel and the lower channel are communicated with diluent through valves, and the diluent passing through the upper channel and the lower channel can be called uncontaminated liquid; the middle port straight-through valve is then led to a pump and then discharged to be waste liquid, the middle port is also provided with an electrode (the electrode is made of outer electrode stainless steel, the inner electrode is made of platinum in a front pool), under normal conditions, the middle liquid is negative pressure, the liquid of the bacteria sample to be counted can enter the rear pool from the front pool, counting is completed in the process of passing through the gem hole, after the bacteria sample to be counted is counted, the rear pool is cleaned in a mode that the liquid enters a liquid inlet at the upper end and the lower end of the rear pool and flows out from a liquid outlet at the other end of the rear pool, for example, the rear pool is cleaned in a mode that the liquid enters the rear pool through a liquid inlet and outlet port of the rear pool, for example, in a mode of liquid inlet and outlet, the liquid is waste liquid, or the liquid sample and the diluent can be contained, the upper channel and the lower channel are connected with each other and are 1 in 2, and 1 is a main channel leading, 2 are respectively connected with the upper and lower channel ports of the rear pool and the middle channel, namely the channel with the electrode.

Under the condition of hole plugging: and (3) closing the negative pressure at the middle liquid port of the rear pool, and optionally applying positive pressure through a pressure pump to enable the rear pool to generate pressure to recoil the gem holes so as to eliminate the complete blockage phenomenon of the gem holes or the incomplete blockage phenomenon of the gem holes. Alternatively, the liquid is fed through the upper and lower liquid ports of the rear tank, so that the rear tank generates pressure to back flush the gem holes, and the complete blockage phenomenon of the gem holes or the incomplete blockage phenomenon of the gem holes is eliminated.

Further, the above counting cell assembly may further optionally comprise:

and the recoil component is used for recoiling when the gem holes are blocked so as to eliminate the blockage of the gem holes. Further, the negative pressure of the liquid in the rear pool is closed, and the liquid is respectively fed from the two liquid ports of the rear pool, so that the rear pool generates pressure to recoil the gem hole, and the blockage of the gem hole is eliminated. Or applying positive pressure by a pressure pump to generate pressure in the rear pool to recoil the gem holes, thereby eliminating the blockage of the gem holes.

Further, the above counting cell assembly may further optionally comprise:

and the burning assembly is used for burning and eliminating the blockage of the gem hole when the blockage phenomenon occurs to the gem hole.

Further, the burning assembly is used for providing a voltage higher than a preset voltage value to the gem hole through the electrode when the blocking phenomenon occurs to the gem hole so as to melt the blocking substance in the gem hole.

After the computer connected with the bacteria counting device detects the hole blockage, namely the alarm or prompt information of the computer, the computer can artificially execute high-voltage ignition to eliminate the hole blockage, namely, the operation button on the computer (PC end) is artificially clicked, a high-voltage ignition circuit is started, namely, direct current voltage (relative low-voltage part) is normally counted, direct current high voltage is generated during ignition, the ignition mode is high-voltage and low-voltage rapid switching, high frequency can be formed in the high-voltage ignition process, arc discharge can be generated on two sides of a jewel hole at the moment of power on and power off, and generated electric sparks just burn hole blockage substances in the jewel hole. Another optional way of burning to eliminate the hole blockage is that when normal counting is to provide stable low-voltage partial pressure through a switch circuit, direct current high voltage is used for burning, and because the high voltage is used for burning, the liquid to be detected is heated and boiled, and protein components are melted and eliminated to achieve the effect of burning to eliminate the hole blockage.

Further, the voltage of the predetermined voltage value is 90 volts to 110 volts.

Further, the voltage of the predetermined voltage value is 110 v.

Further, the forebay is made of plastic materials.

Further, the forebay is made of polyformaldehyde material.

Further, the rear tank is made of plastic materials.

Further, the rear pool is made of a polyformaldehyde material.

The plastic material, especially polyoxymethylene material machine working property is good, guarantees the size of above-mentioned front pool and above-mentioned rear pool easily, and the structure is more firm.

The technical scheme provided by the embodiment of the invention has the following beneficial effects:

1) the embodiment of the invention realizes the automatic application of the device for measuring the number of bacteria by using a resistance counting method, solves the problems of slow time and low efficiency of the existing bacteria counting, and realizes the effects of high speed and accuracy of bacteria counting.

2) The improved jewel holes of the embodiment of the invention ensure that bacteria can pass through the micropores one by one, prevent the overlapping phenomenon from influencing the measurement of the number of the bacteria, realize the measurement of the number of the bacteria by adopting a resistance counting method, and have accuracy and high efficiency; the device is additionally provided with a high-pressure recoil and firing function to prevent the hole from being blocked, if the hole is blocked, the rear pool part is additionally provided with a high-pressure recoil design to eliminate the complete blocking phenomenon of the gem hole or the incomplete blocking phenomenon of the gem hole, if the high-pressure recoil fails, the firing function can be selected to eliminate the hole, namely, the complete blocking phenomenon of the gem hole or the incomplete blocking phenomenon of the gem hole is ensured not to be easily caused under the condition that the aperture is reduced.

3) A bacteria counting signal conditioning circuit is designed in a targeted manner; and a signal conditioning circuit is added, so that non-bacterial signals are filtered, signals of bacterial characteristics are accurately identified, and the condition of misjudgment is reduced.

4) And the sample is uniformly mixed twice before and after the sample is sucked, so that the samples to be detected are fully mixed, and the counting of the number of bacteria in the detection process is more accurate.

5) The blending operation is in the equipment, firstly, other auxiliary blending equipment is not used, and the pollution rate is reduced.

Drawings

The above and other objects, features and advantages of exemplary embodiments of the present invention will become readily apparent from the following detailed description read in conjunction with the accompanying drawings. Several embodiments of the invention are illustrated by way of example, and not by way of limitation, in the figures of the accompanying drawings and in which:

FIG. 1 schematically shows a complete schematic view of a bacteria counting apparatus according to an embodiment of the present invention;

FIG. 1-1 schematically illustrates a schematic view of a complete machine of a bacteria counting apparatus according to an embodiment of the present invention during a sample drawing process;

FIGS. 1-2 schematically illustrate a schematic view of a bacteria counting device as a whole for adding a sample to be tested to be aspirated to a counting cell assembly according to an embodiment of the invention;

FIG. 2 schematically illustrates a structural schematic of a counting cell assembly according to an embodiment of the invention;

FIG. 2-1 schematically illustrates a partially symmetrical cross-sectional structural view of FIG. 2, in accordance with an embodiment of the present invention;

FIG. 3 schematically illustrates a structural schematic of a sampling assembly according to an embodiment of the present invention;

FIG. 3-1 schematically illustrates a cross-sectional, structural schematic view of a sampling needle and swab mating relationship, in accordance with an embodiment of the present invention;

FIG. 3-2 schematically shows a schematic structural view of a reagent plate according to an embodiment of the present invention;

FIG. 4 schematically illustrates a schematic diagram of the operating principle of a resistance counting according to an embodiment of the present invention;

FIG. 4-1 schematically illustrates a schematic liquid path diagram operation of a bacteria counting device according to an embodiment of the present invention;

FIG. 5 schematically illustrates a flow diagram of a signal conditioning circuit according to an embodiment of the present invention;

fig. 6 schematically shows a schematic diagram of the movement of a plunger pump according to an embodiment of the invention.

Detailed Description

The principles and spirit of the present invention will be described with reference to a number of exemplary embodiments. It is understood that these embodiments are given solely for the purpose of enabling those skilled in the art to better understand and to practice the invention, and are not intended to limit the scope of the invention in any way. Rather, these embodiments are provided so that this disclosure will be thorough and complete, and will fully convey the scope of the disclosure to those skilled in the art.

Summary of The Invention

The technical scheme of the invention provides a bacteria counting device, and the bacteria counting device for measuring the number of bacteria by using a resistance counting method is firstly required to be invented, namely a sampling component, a counting cell component and a circuit control system are designed and combined to form a complete bacteria counting device suitable for measuring the number of bacteria by using the resistance counting method.

Exemplary devices

The overall schematic diagram of the bacteria counting device is shown in fig. 1, and the bacteria counting device comprises a counting cell assembly 1, a sampling assembly 2, a signal conditioning circuit 3 and a shell 4. The counting cell component 1 is fixedly connected with the sampling component 2, the signal conditioning circuit 3 is shown in fig. 5, the signal conditioning circuit 3 is arranged below a cableway 41 shown in fig. 1-2, namely, arranged in the bacteria counting device, the signal conditioning circuit 3 is connected with an inner electrode 141 and an outer electrode 142 in the counting cell component 1, the signal conditioning circuit 3 comprises a signal acquisition board, a main control board and the like, the shell 4 is arranged at the outer sides of the counting cell component 1, the sampling component 2 and the signal conditioning circuit 3, the sampling component 2 comprises a movement mechanism, the sampling component 2 acquires bacteria liquid to be detected through the movement mechanism and then puts the bacteria liquid into the counting cell component 1, and the counting cell component 1 is driven by the sampling component 2 to slide on the cableway 41.

The construction of the counting cell assembly 1 is schematically shown in fig. 2, and the partial cross-sectional construction of the counting cell assembly 1 is schematically shown in fig. 2-1, wherein the counting cell assembly 1 includes a jewel hole 11, a front cell 12, a rear cell 13, an inner electrode 141 and an outer electrode 142 communicating the front and rear cells, the jewel hole 11 is located between the front cell 12 and the rear cell 13, and the inner electrode 141 and the outer electrode 142 are connected between the front cell 12 and the rear cell 13.

As shown in fig. 2-1, the rear cell 13 includes an upper liquid port 131, a middle liquid port 132 and a lower liquid port 133, the liquid in the rear cell 13 is under negative pressure, so that the liquid to be tested for bacteria entering the front cell 12 can completely enter the rear cell 13 by passing through the gem hole 11, wherein the effect is most obvious when the middle liquid port 132 of the rear cell 13 is under negative pressure, the upper liquid port 131 and the lower liquid port 133 in the rear cell 13 are two flushing ports, the lead of the outer electrode 142 is screwed on the metal of the outer wall of the middle liquid port 132, and optionally, the inner electrode 141 is platinum for counting bacteria in the bacteria sample to be counted. During detection, a liquid sample to be detected passes through the micropores of the gem hole, and the front and rear pool electrodes sense resistance change, so that a pulse signal is generated in a circuit, and the number of bacteria is measured according to the number of pulses.

The intensity of the measuring signal of the inner electrode 141 and the outer electrode 142 is a sensor for counting bacteria. Because the diluent has conductivity, when a certain voltage is applied between the two electrodes, a certain resistance exists between the micropores of the gem hole 11, and the cells have non-conductivity, when the cells enter the pores, the resistance between the pores can be changed, so that a pulse signal is generated in the circuit, the pulse signal is processed and transmitted to the PC end for analysis, parameters such as the number, the size and the like of the cells can be measured and counted according to the characteristics such as the number of the pulses, the pulse amplitude and the like, the working principle diagram is shown as figure 4, the bacterial number of the bacterial liquid to be measured is obtained through the resistance counting of the bacterial counting device, and the bacterial number is transmitted to the PC (computer) end.

The schematic configuration of the sampling unit 2 is shown in fig. 3, and as shown in fig. 1, 1-1 and 3, the sampling unit 2 includes a three-dimensionally movable mechanical arm, a sampling needle 22, a swab 23, a reagent plate 24, a plunger pump 25, and the like. The movement mechanism included in the sampling assembly 2 includes the mechanical arm and the sampling needle 22, the mechanical arm includes a mechanical arm 21-1 moving along an X-axis, a mechanical arm 21-2 moving along a Y-axis, and a mechanical arm 21-3 moving along a Z-axis, one end of the sampling needle 22 passes through the swab 23, a partial cross-sectional view of a structural schematic diagram of a matching relationship between the sampling needle 22 and the swab 23 is shown in fig. 3-1, when cleaning is required, water is fed from a water inlet pipe 231 and then discharged from a water outlet pipe 232, the other end of the sampling needle 22 is fixedly connected with the mechanical arm 21-2 and moves along with the movement of the mechanical arm 21-2, so as to realize a target sampling function, that is, the sampling needle 22 takes the bacterial test solution from the reagent plate 24, as shown in fig. 1-1, the plunger pump 25 is connected with the sampling needle 22, the plunger pump 25 controls the suction and discharge of the liquid to be tested for bacteria by the sampling needle 22. Two sampling needles 22 are fixed to a holder of a 3-dimensional motion robot, and one swab 23 is provided for each sampling needle 22, or 4 sampling needles 22 may be used.

Moreover, the plurality of sampling needles 22 can be not only relatively stationary but also independently movable.

The schematic structural diagram of the signal conditioning circuit 3 is shown in fig. 5, and the signal conditioning circuit on the signal processing board collects micro signals, and then uploads the number of bacteria through amplification filtering, signal collection and the like.

Fig. 1, 1-1 and 1-2 show a schematic working process of the sampling assembly of the bacteria counting apparatus, when the bacteria counting apparatus is in operation, the sampling assembly 2 moves to a designated position rapidly, the plunger pump 25 controls the sampling needle 22 to mix the bacteria sample solution in the reagent plate 24 and suck the bacteria sample solution, and then the bacteria sample solution is discharged into the forebasin 12 of the counting chamber assembly 1 moving along with the sampling assembly 2.

Specifically, the high-pressure backflushing design is to close the negative pressure of the middle liquid port 132 of the rear tank 13 to feed liquid into the upper liquid port 131 and the lower liquid port 133 of the rear tank 13, and the rear tank 13 generates pressure to backflush the gem hole 11 to eliminate the complete blockage of the gem hole 11 or the incomplete blockage of the gem hole 11, such as high-pressure backflushing failure, and to select a burning function to eliminate the blockage, i.e., to ensure that the complete blockage of the gem hole or the incomplete blockage of the gem hole is not easy to occur under the condition of reduced pore diameter.

Optionally, as shown in fig. 2, the forebay 12 is a four-channel integrated structure, and is made of polyoxymethylene material or other plastic material, the distance between two forebay channel openings 121 of the forebay 12 is 18mm, and the internal liquid volume of the forebay 12 is greater than 2.5 ml. The polyformaldehyde material or other plastic materials are adopted, so that the accurate sizes of the front pool 12 and the rear pool 13 are easily ensured, and the structure is more stable.

As an example, during the process of bacteria passing through the jewel aperture, the jewel aperture may be clogged, thereby causing inaccurate counting of bacteria. In order to solve the problem of inaccurate counting of bacteria caused by blockage of the jewel hole, the embodiment of the invention also provides a detection scheme and a removal scheme of the blockage of the jewel hole, as shown in an optional complete machine diagram of fig. 1-1, wherein a plunger pump is arranged beside the bacteria counting device and is used for high-pressure recoil to remove the blockage of the jewel hole, or/and high-frequency counting voltage is applied to electrodes at two ends of the jewel hole and is used for high-pressure burning to remove the blockage of the jewel hole, and the specific operation is shown in an exemplary method.

Optionally, the plunger pump 25 shown in fig. 1-1 is an important device of the hydraulic system, and as shown in fig. 6, the volume of the sealed working cavity changes to absorb hydraulic fluid by the up-and-down reciprocating motion of the plunger in the cylinder body, so as to mix the liquid to be measured. When the plunger moves upwards, the internal volume of the cylinder body is reduced, liquid in the liquid path is pushed to flow out, when the plunger moves downwards, the internal volume of the cylinder body is increased, the liquid is sucked into the liquid path, and optionally, a gas pump or other pumps can replace the plunger pump 25.

The bacteria counting device provided by the embodiment of the invention further comprises a pump which is connected with the sampling needle through a liquid path and is used for uniformly mixing the bacteria sample to be counted, wherein the bacteria sample to be counted is sucked by the sampling needle, when the pump works in a first mode, the liquid in the liquid path is pushed to flow to the sampling needle, and when the pump works in a second mode, the liquid in the sampling needle is sucked into the liquid path.

Alternatively, the sampling needle 22 may be a plurality of sampling needles, the pump may be connected to each of the sampling needles through a plurality of the liquid lines, the pump may be a plunger pump or a gas pump, and the connection manner of the sampling needle 22 and the plunger pump 25 is shown in fig. 1-1, for example.

Optionally, the pump is the plunger pump, wherein the plunger pump includes: a cylinder body; and a plunger which reciprocates up and down in the cylinder, wherein when the plunger moves upward, an inner volume of the cylinder is decreased to push the liquid in the liquid path to the sampling needle, and when the plunger moves downward, the inner volume of the cylinder is increased to suck the liquid in the sampling needle into the liquid path, as shown in fig. 6.

Exemplary method

Optionally, two platinum electrodes are respectively arranged on two sides of the laser-formed gem hole, and because the diluent has conductivity, when a certain voltage is applied between the two electrodes, a certain resistance is arranged between the micropores. When cells enter the pores, the resistance among the pores is changed, so that a pulse signal is generated in the circuit, the pulse signal is processed and transmitted to a PC (personal computer) end for analysis, and parameters such as the number, the size and the like of the cells can be measured and counted according to the number of the pulses, the pulse amplitude and other characteristics.

A bacterial count signal conditioning circuit and an acquisition algorithm are designed in a targeted manner, effective signals are completely reserved through amplifying the signals, and the effective signals are adjusted to the amplification times which are most beneficial to algorithm identification through adjusting gains; the low pass filtering filters out high frequency noise and the out-of-limit amplitude by buffering clipping. The bacterial characteristic signals are accurately identified through algorithms such as pulse identification, slope identification, peak detection, trough detection, broadband detection and the like, so that the bacterial quantity is obtained from the pulse signals.

In the event that said pulse signals comprise a first type of set of pulse signals, said circuit control system or processing device determining a first number of said first type of set of pulse signals, wherein each pulse signal of said first type of set of pulse signals is a pulse signal triggered by one of said bacteria through said gemstone aperture; in the case where the pulse signal includes a second type of pulse signal, the circuit control system or the processing device determines a second number as a product of a number of the second type of pulse signal and a predetermined number, wherein each of the second type of pulse signal is a pulse signal generated by the predetermined number of bacteria simultaneously passing through the jewel hole.

Determining the number of bacteria in said sample of bacteria to be counted as said first number in case said pulse signals comprise only one set of pulse signals of said first type; determining the number of bacteria in said sample of bacteria to be counted as said second number in case said pulse signals comprise only one set of pulse signals of said second type; in the case where the pulse signal includes a set of pulse signals of the first type and a set of pulse signals of the second type, the number of bacteria in the bacterial sample to be counted is determined as the sum of the first number and the second number.

As an example, the circuit control system or the processing device in the embodiment of the present invention may determine whether the pulse signal includes a group of pulse signals of the second type by:

when only 1 bacterium can pass through, the counting is accurate, 2 bacteria or 3 bacteria simultaneously pass through a second type pulse signal generated by the gem pore, when the second type pulse signal and the first type pulse signal are within an error range, the second type pulse signal is recorded as effective counting, otherwise, an error is reported for recounting or a counting result is converted according to the error value.

As an example, during the process of bacteria passing through the jewel aperture, the jewel aperture may be clogged, thereby causing inaccurate counting of bacteria. In order to solve the problem of inaccurate counting of bacteria caused by blockage of the jewel hole, the embodiment of the invention also provides a detection scheme and a removal scheme of the blockage of the jewel hole.

As an exemplary detection scheme for gem hole blockage, the embodiment of the invention further comprises: determining that the gem hole is blocked when the voltage between the front pool and the rear pool is detected to exceed a predetermined threshold, wherein the front pool is an anode and the rear pool is a cathode, the more the blockage of the gem hole is serious, the larger the resistance between two sides of the gem hole is, and the larger the voltage between the front pool and the rear pool is, and the predetermined threshold can be set according to different measurement requirements (for example, different measurement precision) of the number of bacteria.

As an exemplary solution for removing the plugged jewel hole by recoil, the embodiment of the present invention further includes:

as an exemplary solution for eliminating the plugged hole by burning, the embodiment of the present invention further includes: high pressure recoil and high pressure firing.

If meet the plug hole, the middle part is just to the jewel hole position, from the middle flushing opening malleation recoil liquid, two upper and lower flushing openings discharge, the combination control of plunger pump and valve, through last lower liquid mouth to exerting the malleation in the pond of back, the waste liquid is discharged from preceding pond waste liquid mouth, the waste liquid draws through the cooperation of battery valve and waste liquid pump.

The firing process is under 110V voltage, high-frequency counting voltage is added on the electrodes at two ends of the small hole, during normal counting, the counting voltage is direct current voltage continuously provided, during high-voltage firing, the high-frequency counting electrode is designed to be powered on and powered off at short intervals, so that high frequency is formed, at the moment of power on and power off, arc discharge is generated between the two electrodes, the emitting point of electric sparks is the small hole, so that protein and fragments are easily removed, other instruments are designed to be independently powered by alternating current, and the front end of an electrode wire is controlled by a relay or a silicon controlled rectifier. Optionally, the method can also be used for burning and eliminating the hole blocking phenomenon by boiling and heating the protein under high pressure to melt the protein.

Optionally, with a multi-probe design and a novel probe structure, the two sampling needles 22 move simultaneously, so that the effect of 4 sampling needles is achieved, and the cost is saved; the design is applied to counting bacteria, 2 stainless steel sampling needles are driven by a 3-dimensional motion mechanical arm, as shown in figure 3, the interval between two sampling needles 22 is 18mm, a target plate is removed for sample suction, liquid to be measured is spitted into two counting cells, 4 counting cells are provided in total, and at the moment, the two counting cell channels start to work; and then the 3-dimensional motion mechanical arm drives 2 stainless steel sampling needles to remove the target plate for sample suction, the liquid to be detected is spit into the other two counting cells, at the moment, the two counting cell channels start to work, the sample suction is removed again until the channel detection is finished, and the same actions are repeated. The simultaneous movement of the two sampling needles 22, and the multi-channel detection, can save both the waiting time and the cost of the probe assembly.

Optionally, the three-dimensional arm moves together with the counting cell, so that the time from sample suction to sample delivery is shortened.

The three-dimensional arm and the counting cell move together and are relatively static, and when the sample needs to be detected, the three-dimensional arm X, Y, Z shaft is directly used for nearby lofting to the counting cell after the sample is sucked, and excessive 3-dimensional motion is not needed.

Optionally, as shown in fig. 3, the sampling needles 22 are spaced at 18mm intervals, and as shown in a schematic structural diagram of the reagent plate 24 shown in fig. 3-2, the distance between two test hole sites 241 of the reagent plate 24 is 9mm, the distance between two sampling needles 22 is exactly 2 times of the distance between the test hole sites of the reagent plate, so that the stroke is short, time is saved, and it is avoided that the movement stroke is lengthened to increase the operation time and reduce the efficiency.

Optionally, as shown in fig. 4, after the liquid is injected into the forebay by the sample adding needle, the forebay liquid is brought to the rear bay by the negative pressure action, and then passes through the jewel hole, when bacteria pass through the jewel hole voltage, pulse signals can be generated (a constant current source is provided, the bacteria change through representing resistance and then the voltage changes), the circuit filters and amplifies the signals, then the signals reach the single chip microcomputer after filtering, the single chip microcomputer performs the process of AD sampling, the program of the single chip microcomputer also has a pulse recognition algorithm, and after the processing, the signals can be uploaded to the PC software. And the process of processing the signals comprises the following steps: the AD samples are sampled approximately in the order of 10M and then the algorithm processes to a number of K, i.e. total, histogram information. And then upload to the PC.

Optionally, in the working process of the bacteria counting device, the flow chart of the liquid flow direction is as shown in fig. 4-1, the bacteria counting device is a 4-channel counting liquid-path bacteria counting device, because the liquid path of each channel is consistent, the working principle of the bacteria counting device is described by taking two channels 1 and 2(CH1 and CH2) as an example: as shown in the liquid path diagram of fig. 4-1, before sample adding and counting, before each counting, the counting cell module 1 is cleaned, the V1 solenoid valve is matched with the 10ML pump to suck up diluent (bacterial reagent to be measured), then the liquid is injected into the anterior cell 12 through the matching of the V1 solenoid valve, the V2 solenoid valve, the V3 solenoid valve, the V4 solenoid valve and the pump, then the positive pressure is applied to the liquid path to flush the gem pore 11 through the matching of the 10ML pump with the V1 solenoid valve, the V2 solenoid valve and the V3 solenoid valve, then the waste liquid in the anterior cell 12 is completely discharged through the V8 solenoid valve, the V9 solenoid valve and the P1 pump, and the P3 pump, the V6 solenoid valve and the V7 solenoid valve discharge the waste liquid in the posterior cell 13. The sample adding and counting process is as follows: adding a liquid into the forebay 12, then adding a diluent through a combination of a solenoid valve and a pump for dilution, lifting the swab 23, and cleaning the swab 23: the sampling needle 22 is lifted, the lower bottom surface of the sampling needle 22 is wrapped in the swab 23, the V5 solenoid valve is matched with the P1 pump to discharge the diluent which washes the outer wall of the sampling needle 22 from the V4 solenoid valve channel to a waste liquid pool, and then the V5 solenoid valve is matched with the P1 pump to discharge the diluent which washes the inner wall of the sampling needle 22 from the V4 solenoid valve channel to the waste liquid pool. The V4 electromagnetic valve is 3-way, one inlet, two outlets (assumed as 1 and 2), at least one outlet is communicated with the inlet at the same time, so that the diluent can be controlled to clean the inner wall and the outer wall of the sampling needle 22, and the liquid to be detected is sucked after the cleaning is finished. And the two channels which load the sample first start to count for a certain time through the negative pressure of the V6 electromagnetic valve and the P3 pump, when the preset time is reached, the front pool 12 and the rear pool 13 are cleaned through the matching of the respective electromagnetic valve and the pump, and the next sample injection is waited.

The embodiment of the invention also provides a bacteria sampling mode, which comprises the following steps: driving a sampling needle to suck a bacterial sample to be counted from a reagent plate through a three-dimensional motion mechanical arm; uniformly mixing the bacteria sample to be counted in the sampling needle by a pump, wherein the pump is connected with the sampling needle through a liquid route, when the pump works in a first mode, the liquid in the liquid route is pushed to flow to the sampling needle, and when the pump works in a second mode, the liquid in the sampling needle is sucked into the liquid route; the sampling needle is driven to move to the counting cell assembly through the three-dimensional motion mechanical arm; and controlling the sampling needle to add the sucked bacterial sample to be counted into the counting cell assembly.

The comparison of the data of the number of bacteria measured by whether the test agent is mixed by a pump before the sample is sucked is shown in the following table:

the test results show that the number of bacteria obtained by uniformly mixing the test agent to be tested by using the pump before sample suction is obviously higher than that obtained by uniformly mixing the test agent to be tested by using no pump before sample suction, namely, the number of bacteria obtained by uniformly mixing the test agent to be tested by using the pump before sample suction is closer to the objective number of bacteria.

It should be noted that although in the above detailed description several units/modules or sub-units/modules of the apparatus are mentioned, such a division is merely exemplary and not mandatory. Indeed, the features and functionality of two or more of the units/modules described above may be embodied in one unit/module according to embodiments of the invention. Conversely, the features and functions of one unit/module described above may be further divided into embodiments by a plurality of units/modules.

Moreover, while the operations of the method of the invention are depicted in the drawings in a particular order, this does not require or imply that the operations must be performed in this particular order, or that all of the illustrated operations must be performed, to achieve desirable results. Additionally or alternatively, certain steps may be omitted, multiple steps combined into one step execution, and/or one step broken down into multiple step executions.

While the spirit and principles of the invention have been described with reference to several particular embodiments, it is to be understood that the invention is not limited to the disclosed embodiments, nor is the division of aspects, which is for convenience only as the features in such aspects may not be combined to benefit. The invention is intended to cover various modifications and equivalent arrangements included within the spirit and scope of the appended claims.

Claims

1. A bacteria sampling device, comprising:

the sampling assembly comprises a three-dimensional motion mechanical arm and a sampling needle, the three-dimensional motion mechanical arm is used for driving the sampling needle to suck a bacterial sample to be counted from the reagent plate, and the sampling needle is used for adding the sucked bacterial sample to be counted into the counting cell assembly;

the pump, through the liquid route with the sampling needle is connected, is used for the mixing the sampling needle inhales and gets treat count bacterium sample, wherein, work as the pump is when working with first mode, promotes the liquid flow in the liquid route flows to the sampling needle, works as the pump is when working with the second mode, will liquid in the sampling needle is inhaled the liquid route.

2. The device of claim 1, wherein the sampling needle is a plurality of sampling needles, the pump is connected to each sampling needle through a plurality of the liquid lines, and the pump is a plunger pump or a gas pump.

3. The apparatus of claim 1, wherein the apparatus further comprises:

the plate-type plate-making machine comprises a machine frame, wherein a plate groove is formed in the machine frame;

the reagent plate is clamped in the plate groove;

wherein the pump is disposed inside the housing.

4. The apparatus of claim 1, wherein the pump is the plunger pump, wherein the plunger pump comprises:

a cylinder body;

and a plunger which reciprocates up and down in the cylinder, wherein when the plunger moves upward, an internal volume of the cylinder becomes smaller to push the liquid in the liquid path to flow to the sampling needle, and when the plunger moves downward, the internal volume of the cylinder becomes larger to suck the liquid in the sampling needle into the liquid path.

5. The device of claim 4, wherein the plunger is further configured to move up and down in the cylinder to mix the bacteria sample to be counted in the reagent plate before the sampling needle contacts the bacteria sample to be counted in the reagent plate but does not aspirate the bacteria sample to be counted.

6. The apparatus of any of claims 1 to 5, further comprising:

a counting cell assembly for performing bacterial counting on the bacterial sample to be counted.

7. A bacteria sampling method, comprising:

driving a sampling needle to suck a bacterial sample to be counted from a reagent plate through a three-dimensional motion mechanical arm;

uniformly mixing the bacteria sample to be counted in the sampling needle by a pump, wherein the pump is connected with the sampling needle through a liquid route, when the pump works in a first mode, the liquid in the liquid route is pushed to flow to the sampling needle, and when the pump works in a second mode, the liquid in the sampling needle is sucked into the liquid route;

the three-dimensional motion mechanical arm drives the sampling needle to move to the counting cell assembly;

and controlling the sampling needle to add the sucked bacterial sample to be counted into the counting cell assembly.

8. The method of claim 7, wherein the pump comprises: cylinder body and plunger, wherein, it is right through the pump in the sampling needle treat count bacterium sample and carry out the mixing, include:

and controlling the plunger to reciprocate up and down in the cylinder, wherein when the plunger moves upwards, the internal volume of the cylinder is reduced to push the liquid in the liquid path to flow to the sampling needle, and when the plunger moves downwards, the internal volume of the cylinder is increased to suck the liquid in the sampling needle into the liquid path.

9. The method of claim 7, wherein prior to said drawing the sample of bacteria to be enumerated from the reagent plate by the three-dimensional motion of the robotic arm, the method further comprises:

and before the sampling needle contacts the bacterial sample to be counted in the reagent plate but does not suck the bacterial sample to be counted, uniformly mixing the bacterial sample to be counted in the reagent plate through the pump.

10. The method of claim 8, wherein prior to said drawing the sample of bacteria to be enumerated from the reagent plate by the three-dimensional motion of the robotic arm, the method further comprises:

and controlling the plunger to move upwards and downwards in the cylinder body so as to uniformly mix the bacteria sample to be counted in the reagent plate.