CN111893034A - Bacteria counting device with conduction heating mechanism - Google Patents

Abstract

The invention provides a heating device for bacterial counting, comprising: the heating plate is used for placing a reagent plate, and the reagent plate is used for containing a bacterial sample to be counted; a heating block, wherein the heating plate is positioned above the heating block; a heating film, wherein the heating film is in contact with the heating block; a temperature sensor connected to the heating film; and the heating control board is connected with the temperature sensor and the heating block and is used for carrying out temperature control on the heating block according to the temperature parameters acquired by the temperature sensor. The required target temperature is reached within 5 minutes, the heating time is reduced, the whole incubation time is saved, the heating part is directly attached to the heated part, and the heating is rapidly carried out; the reagent board and the heating plate are in a laminating design, the gap is smaller than 0.1mm, and the heating effect and the heat preservation effect are controlled more effectively and accurately.

Description

Bacteria counting device with conduction heating 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 conduction heating 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. This is also the basis of design that is commonly used by most hematology analyzers.

However, the bacteria counting device in the prior art has the following problems:

the design is applied to counting bacteria, and the current counting instrument has the following problems:

first, no device for measuring the number of bacteria by using a resistance counting method exists in the market at present.

Secondly, the existing market utilizes the design to be based on equipment for blood analysis, and the equipment does not have an incubation function.

And thirdly, the existing counting equipment has no function of independently counting the incubation time for each plate.

The traditional device has no requirement of temperature incubation, and the temperature control is not from the point of view, because the design is aimed at counting bacteria, and the step of culturing the bacteria is needed before detection due to the specificity of the detection object. Because the test time is relatively short (2h), there is not enough time to slowly wait for the test temperature to approach the equilibrium, and the required test temperature is required to be reached immediately at the beginning of the test, so the temperature control is important. Therefore, how to design a heating mechanism of the automatic resistance counting device for bacteria with high speed and high efficiency and how to heat the heating mechanism are technical problems to be solved by the design.

Disclosure of Invention

The embodiment of the invention provides a bacteria counting device, which at least solves the technical problems that the micro-particle counting device in the prior art cannot rapidly and accurately measure the number of bacteria and how to heat the bacteria.

An embodiment of the present invention provides a heating device for counting bacteria, including:

the heating plate is used for placing a reagent plate, and the reagent plate is used for containing a bacterial sample to be counted;

a heating block, wherein the heating plate is positioned above the heating block;

a heating film, wherein the heating film is in contact with the heating block;

a temperature sensor connected to the heating film; and

and the heating control board is connected with the temperature sensor and the heating block and is used for carrying out temperature control on the heating block according to the temperature parameters acquired by the temperature sensor.

Optionally, the heating plate is attached to the upper surface of the heating block.

Optionally, a heating groove is arranged in the heating plate, the heating groove is matched with a reagent hole on the reagent plate for containing the bacteria sample to be counted, and the heating groove is used for containing the reagent hole.

Optionally, the interval between the bottom of the heating groove and the heating block is smaller than a first threshold.

Optionally, the heating film is located between the heating plate and the upper surface of the heating block.

Optionally, the heating block includes a set of heating components arranged according to a predetermined pattern for generating heat.

Optionally, the heating block further includes: the heat conduction material wraps the heating component.

Optionally, the thickness of the portion of the heat conductive material located above the upper surface of the heating element is greater than a predetermined value in a range of 2mm to 5 mm.

Optionally, the heating element is located at a position corresponding to a position of a heating groove in the heating plate, wherein the heating groove is matched with a reagent hole in the reagent plate for containing the bacterial sample to be counted, and the heating groove is used for accommodating the reagent hole.

Optionally, the heating control plate includes: and the processor is used for increasing the temperature of the heating block to a second threshold value within a preset time period and stabilizing the temperature at the second threshold value.

Optionally, the present invention provides a bacteria counting apparatus comprising: the heating device of any one of the above; 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; the sampling assembly is arranged on the rack and is used for acquiring the bacterial sample to be counted from the reagent plate; and the counting cell assembly is arranged on the rack and used for counting bacteria in the bacteria sample to be counted.

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) The first time, the incubation mode is applied to bacteria counting equipment, so that the automatic incubation and counting can be realized, and the non-full-automatic application in the prior art is solved.

5) The PID control algorithm is combined with the solid block heating mode, the required target temperature is achieved within 5 minutes, the heating time is reduced, and the whole incubation time is saved.

6) The heating part is directly attached to the heated part, and the temperature is rapidly increased; the reagent board and the heating plate are in a laminating design, the gap is smaller than 0.1mm, and the heating effect and the heat preservation effect are controlled more effectively and accurately.

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;

3-3 schematically illustrate a structural schematic of a heating assembly according to an embodiment of the present invention;

FIGS. 3-4 schematically illustrate a structural schematic of a heating block according to an embodiment of the present invention;

FIGS. 3-5 schematically illustrate a schematic diagram of a 48-well reagent plate according to an embodiment of the present invention;

FIGS. 3-6 schematically illustrate a detailed flow diagram of a temperature control 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 shows a flow diagram of a signal conditioning circuit according to an embodiment of the present 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, which measures the number of bacteria by using a resistance counting method, firstly needs to invent the bacteria counting device which measures the number of bacteria by using the resistance counting method, namely, a sampling component, a counting cell component and a circuit control system are designed and combined to be suitable for measuring the number of bacteria by a resistance counting method, on the basis, the invention provides a bacteria counting device with a conduction heating mechanism, an automatic resistance counting device heating mechanism aiming at bacteria, the coordination of heating and counting, for example, the heating time and temperature (automatic type) correspond to the counting, because bacteria need to be counted quickly and accurately, the requirement of quick culture of the bacteria needs to be met, the heating time and temperature are automatically controlled according to different samples, so that subsequent counting service is served, and the accuracy and timeliness are improved. The hot melting is large and quick, the heat can be stored in time, and the thickness of the plate is more than 5 mm; the contact is tight, the contact area between the sample box and the heating plate is large, and the power is large.

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 probes can not only be relatively static, but also can move independently.

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 elimination 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 eliminate the blockage, or/and high-frequency counting voltage or direct-current high voltage is applied to electrodes at two ends of the small hole and is used for high-pressure burning to eliminate the blockage.

Optionally, the conductive heating mechanism is mainly a heating assembly, as shown in fig. 3-3, the heating assembly is composed of a heating block, a detection plate, a heating film, a temperature sensor, and a heating control plate, and the heating block is shown in fig. 3-4. The heating plate is used for placing a reagent plate, and the reagent plate is used for containing a bacterial sample to be counted; a heating block, wherein the heating plate is positioned above the heating block; a heating film, wherein the heating film is in contact with the heating block; a temperature sensor connected to the heating film; and the heating control board is connected with the temperature sensor and the heating block and is used for carrying out temperature control on the heating block according to the temperature parameters acquired by the temperature sensor.

Optionally, the hardware conduction mode adopts solid block heating with large specific heat capacity, and the solid block adopts an aluminum block.

Optionally, in the embodiment of the present invention, a bonding design and a tight connection mode are adopted, a heating film is arranged below the solid block, the bottom plate is pressed and tightly contacted, a clamping groove for clamping the reagent plate is arranged on the solid block, and the 48-hole reagent plate shown in fig. 3-5 can be placed in a clamping groove hole corresponding to the heating assembly shown in fig. 3-3.

The embodiment of the invention is characterized in that the heat capacity of the heating block is large, the distance between the detection plate and the heating block is small, and the joint design is easy for heating and heat preservation.

As shown in fig. 3-4, the thickness of the heating block is larger than 2mm, the distance is larger than 0.3mm, the heating film is pasted below the heating plate and is in direct contact, the model design of the reagent plate is in fit design with the heating block, the wall thickness of the reaction cup of the reagent plate is smaller than 0.5mm, the gap between the reaction cup and the heating plate is smaller than 0.1mm, and the reaction cup is a container, namely the reaction cup, for containing the liquid to be tested for bacteria in the reagent plate shown in fig. 3-5.

Optionally, the temperature software control mode is PID control, the required target temperature is reached within 5 minutes, the heating time is tested by adopting a PID temperature rise mode, and the specific flow of the temperature control is shown in fig. 3-6.

Exemplary method

The time data for measuring the temperature rise by adopting the technical scheme of the embodiment is shown in the following table:

1) when the ambient temperature is 10 ℃, the temperature of the reagent plate, i.e., the temperature of the liquid to be tested for bacteria, is measured using the bacteria counting apparatus having the conductive heating mechanism shown in this embodiment.

Time(s)	Target temperature (37 ℃ C.)
		0	10℃
30	15℃
		60	19℃
90	24℃
		120	27℃
150	29℃
		180	33℃
210	36℃
		240	37℃

2) When the ambient temperature is 10 ℃, the temperature of the reagent plate, i.e., the temperature of the liquid to be tested for bacteria, is measured using the bacteria counting apparatus having the conductive heating mechanism shown in this embodiment.

Time(s)	Target temperature (37 ℃ C.)
		0	25℃
30	26℃
		60	28℃
90	31℃
		120	33℃
150	36℃
		180	37℃
210	37℃
		240	37℃

From the above experiment, it can be known that the temperature of the liquid to be tested for bacteria can be rapidly increased by using the bacteria counting device with the conduction heating mechanism shown in this embodiment, so as to achieve the rapid goal.

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.

Since the incubation time is equal to the temperature stabilization process time + the temperature rise time, the temperature rise time may be different due to the external environment temperature, but the temperature can reach the predetermined temperature within a certain time. The incubation temperature is closely related to the final number of bacteria during growth, the number counting which cannot meet the requirement cannot be accurate (because the instrument has certain sensitivity, the counting can be more accurate only by reaching certain magnitude, or the counting cannot be detected too little), if the instrument has certain sensitivity, the temperature can be cultured for 4 generations at about 37 ℃, and if the temperature is 30 ℃, the seed can be cultured for 2 generations. Therefore, the time and temperature required for heating must be as fast as possible to minimize the effect on bacterial growth during the time period of temperature rise.

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.

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 (10)

1. A heating device for bacterial enumeration, comprising:

the heating plate is used for placing a reagent plate, and the reagent plate is used for containing a bacterial sample to be counted;

a heating block, wherein the heating plate is positioned above the heating block;

a heating film, wherein the heating film is in contact with the heating block;

a temperature sensor, wherein the temperature sensor is connected to the heating film; and

and the heating control board is connected with the temperature sensor and the heating block and used for carrying out temperature control on the heating block according to the temperature parameters acquired by the temperature sensor.

2. The apparatus of claim 1, wherein the heating plate is attached over an upper surface of the heating block.

3. The apparatus of claim 1, wherein the heating plate is provided with a heating groove therein, the heating groove is matched with a reagent hole on the reagent plate for containing the bacterial sample to be counted, and the heating groove is used for accommodating the reagent hole.

4. The apparatus of claim 3, wherein a spacing between a bottom of the heating well and the heating block is less than a first threshold.

5. The apparatus of claim 1, wherein the heating film is positioned between the heating plate and an upper surface of the heating block.

6. The apparatus of claim 1, wherein the heating block comprises a plurality of heating elements arranged in a predetermined pattern for generating heat.

7. The apparatus of claim 6, wherein the heating block further comprises: a thermally conductive material encasing the heating element.

8. The apparatus of claim 7, wherein a thickness of a portion of the thermally conductive material above the upper surface of the heating element is greater than a predetermined value in the range of 2mm to 5 mm.

9. The apparatus of claim 6, wherein the heating element is positioned to correspond to a heating well in the heating plate, wherein the heating well is aligned with a reagent well in the reagent plate for containing the bacterial sample to be counted, and the heating well is configured to receive the reagent well.

10. The apparatus of claim 1, wherein the heat control plate comprises: and the processor is used for increasing the temperature of the heating block to a second threshold value within a preset time period and stabilizing the temperature at the second threshold value.

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