CN111289556A

CN111289556A - Device and method for detecting liquid phase change

Info

Publication number: CN111289556A
Application number: CN201811486535.9A
Authority: CN
Inventors: 苏楠; 和建伟
Original assignee: Beijing Bicheng Biotechnology Co ltd
Current assignee: Beijing Jushu Biotechnology Co ltd
Priority date: 2018-12-06
Filing date: 2018-12-06
Publication date: 2020-06-16
Anticipated expiration: 2038-12-06
Also published as: CN111289556B

Abstract

The invention provides a device for detecting liquid phase change, which comprises: the sample placing area is used for placing a liquid sample to be detected and completing the phase change process of the liquid in the sample placing area so as to detect the liquid sample; the pulse module is used for applying pulse pressure to the sample to be detected; the detection module is used for detecting the change of the pressure in the pipeline on one side of the detection module; the adjusting area is used for adjusting the intensity of a signal which is detected by the detection module and fed back from the sample to be detected; a conduit having an inlet and an outlet for ingress and egress of a sample; the pulse module and the detection module are arranged on two sides of the sample placing area, the detection of the change of the pressure in the pipeline on one side of the detection module means that the detection module detects the change of the pressure, applied to a sample to be detected by the pulse module and transmitted to one side of the detection module through the sample, along with the time, and outputs the change of the pressure along with the time as a signal, and the adjusting area is positioned between the detection module and the sample placing area.

Description

Device and method for detecting liquid phase change

Technical Field

The invention relates to a device for detecting liquid phase change and a method for detecting liquid phase change by using the device.

Background

It is known that blood coagulation in a living heart or blood vessel causes thrombosis and thrombotic diseases, for example, Acute Myocardial Infarction (AMI), cerebral thrombosis, Deep Vein Thrombosis (DVT), Disseminated Intravascular Coagulation (DIC), and the like; extravascular bleeding of blood can lead to bleeding and can cause bleeding disorders such as allergic purpura, thrombocytopenic purpura, hemophilia, and liver disease bleeding. In order to effectively inhibit these diseases, it is necessary to periodically detect the thrombosis in the blood of a patient.

Currently, in the whole blood system, a Thrombus Elastogram (TEG) is a common method for measuring blood coagulation and thrombus formation by using a thromboelastometer, and is an index reflecting dynamic changes in blood coagulation (including a rate of fibrin formation, firmness of a dissolved state and a coagulated state, and elasticity). Thrombelastogram is a graph drawn by a thrombelastometer. The main components of a conventional thromboelastometer include: a stainless steel blood cup capable of automatically adjusting the constant temperature (37 ℃), a small stainless steel cylinder inserted into the cup and a sensor capable of being connected with the cylinder. The blood cup is arranged on a reaction tank which can rotate back and forth at an angle of 4 degrees and 45 degrees, and blood is contained between the cup wall and the cylinder. When the blood sample is in a liquid state, the round-trip rotation of the cup can not drive the cylinder, the signal reflected to the tracing paper by the sensor is a straight line, when the blood begins to coagulate, resistance is generated between the cup and the cylinder due to the adhesion of fibrin, the cylinder is driven to move simultaneously by the rotation of the cup, the resistance is increased along with the increase of the fibrin, the movement of the cylinder driven by the cup is changed, and the signal traces the thrombelastogram formed on the tracing paper by the sensor.

In addition, many other liquid phase transition processes are also detected or attended to, such as the solidification of some polymer solutions, for example, protein solutions, protein curds, gelatin, and polymer polymerization.

Disclosure of Invention

As described above, thromboelastometers are commonly used in the art to measure the coagulation of blood and the formation of thrombi. However, when the thromboelastometer is used for detection, a large amount of blood is often needed, and a large amount of blood needs to be extracted from a patient for detection, which causes great burden to the patient and higher requirements on operation for a clinician.

Therefore, if the detection of blood coagulation can be realized by a small amount of sample, for example, micro-liter, in the field, the burden of the patient and the risk during blood collection can be greatly reduced. In addition, because the detection of blood coagulation needs to avoid the interference of external environment as much as possible in the detection process, a detection device which can realize very small influence on the collected blood in the whole detection process is also needed.

JP2007-271323a discloses a measurement method capable of simultaneously detecting the blood viscosity and the amount of thrombus formation in a short time at low cost. This patent document relates to a blood holding container having an opening to which a capillary tube is connected, a pressurizing device for discharging blood in the blood holding container from the capillary tube at a constant flow rate, and a detecting device for detecting the viscosity of blood and the amount of thrombus formation.

CN102762991A discloses a microchip for platelet measurement and a platelet measurement device using the same. The microchip disclosed in this patent document is a microchip for measuring platelet function by inducing platelet aggregation by flowing blood in a channel, and has a channel provided inside, wherein collagen is at least partially coated in the channel in order to adhere to platelets, a plurality of walls extend in the direction in which blood flows in the channel, and partition the width of the channel to form channel partitions, and the walls are subjected to a treatment for making the surface roughness (Ra) 10 to 200 nm. By using this device, a platelet function of detecting blood using a trace amount of blood can be realized.

CN101292161A discloses a device for monitoring thrombus formation and a method for detecting thrombus formation. Providing a thrombus-inducing agent in at least a portion of the device comprising a thrombus formation chamber; an inlet tube connected to the thrombus formation chamber and through which blood flows into the thrombus formation chamber; and a drug tube connected to the inlet tube and through which a drug for releasing anticoagulation treatment to go or promote blood coagulation is supplied. The method comprises flowing anticoagulated blood into a thrombosis chamber, providing a thrombosis inducing agent that induces thrombosis in at least a portion of the thrombosis chamber while releasing anticoagulation treatment or promoting blood coagulation, thereby monitoring thrombosis.

CN101874208A discloses a microchip and a blood monitoring device. The interior of the microchip comprises: a first channel into which a first liquid selected from whole blood, platelet-rich plasma, and a drug-treated liquid thereof flows; a second flow path connected to the first flow path, into which a second liquid containing a chemical that reacts with the first liquid flows; and a merged channel extending from a connection portion between the first channel and the second channel; the microchip is characterized in that a stirring section having a stirring bar for mixing the first liquid and the second liquid is provided in the merged channel. By using the device, the reaction performance of blood can be detected by effectively mixing trace blood and the medicament.

A cup-based device for blood coagulation measurement and testing is disclosed in CN 102099676A. The device includes a blood clot detection instrument and a cup for the blood clot detection instrument. The cup includes a blood sample receiver inlet and a channel structure, the channel structure including: at least one test channel for making a measurement of blood clotting time; a sampling channel having at least one surface portion that is hydrophilic in communication with the blood sample receiver inlet and the at least one test channel; and a waste channel having at least one surface portion that is hydrophilic in communication with the sampling channel; and a vent opening in communication with the sampling channel. The exposure of the optical sensor activates a pump module of the blood clot detection instrument that draws a desired volume of a blood sample into the at least one test channel.

The above information is only for enhancement of understanding of the background of the invention and therefore it may contain information that does not form the prior art that is well known to those of ordinary skill in the art. The above patent applications and the prior art use various devices of relatively complex construction to perform the detection of minute amounts of blood. In addition, in the above-mentioned apparatus, it is often necessary to perform coagulation inhibition treatment on the surface of the member which comes into contact with blood, for example, surface treatment with heparin, polyacetyl lactone, poly-2-methoxyethyl adenine or the like.

In addition, with respect to other liquid samples other than blood samples, there is a similar problem in that it is desired to realize detection with a minute volume and to avoid interference and influence of the liquid sample to the outside during the entire process of detecting coagulation of the liquid.

Further, it is desirable to achieve continuous, rapid, and efficient detection of different liquid samples without mutual interference and contamination between different samples.

In view of the above circumstances, the present invention is intended to provide a liquid phase change detection apparatus having a simple structure, capable of rapidly detecting a phase change process of a liquid to rapidly obtain information on a liquid phase change, such as thrombus formation, and capable of realizing detection only for a trace amount of a liquid sample, and a method for detecting a liquid phase change using the apparatus.

The purpose of the invention is realized by the following technical scheme.

1. An apparatus for detecting a phase change of a liquid, comprising:

the sample placing area is used for placing a liquid sample to be detected and completing the phase change process of the liquid in the sample placing area so as to detect the liquid sample;

the pulse module is used for applying pulse pressure to the sample to be detected;

the detection module is used for detecting the change of the pressure in the pipeline on one side of the detection module;

the adjusting area is used for adjusting the intensity of a signal which is detected by the detection module and fed back from the sample to be detected;

a conduit having an inlet and an outlet for ingress and egress of a sample;

wherein, the pulse module and the detection module are arranged at two sides of the sample placing area, and the detection of the pressure change in the pipeline at one side of the detection module means that the detection module detects the change of the pressure applied to the sample to be detected by the pulse module and transmitted to one side of the detection module through the sample along with the time, and outputs the change of the pressure along with the time as a signal,

the conditioning zone is located between the detection module and the sample placement zone.

2. The apparatus according to item 1, wherein a cross-sectional shape of the sample placement area satisfies the following condition:

the maximum width of the sample placing area in the sample feeding direction of the sample to be detected is D1, the maximum width of the sample placing area in the direction perpendicular to the sample feeding direction of the sample to be detected is D2, and D1 is more than or equal to D2;

the sample placing areas are basically symmetrical along the sample feeding direction of a sample to be detected; and

the shape of one side of the sample placing area in the sample feeding direction along the sample to be measured is an arc shape which is substantially outward convex.

3. The apparatus according to item 2, wherein the cross-sectional shape of the sample placement area further satisfies the following condition:

1＜D1/D2≤8，

preferably 1 < D1/D2. ltoreq.7,

more preferably 1 < D1/D2. ltoreq.6,

further preferably 1 < D1/D2. ltoreq.5.

4. The apparatus of item 3, wherein the cross-sectional shape of the sample placement area further satisfies the condition:

when the inner diameter of the pipe for the inlet and outlet for the sample to enter and exit is R,

satisfying 2 < D1/R < 24 >,

preferably 2. ltoreq. D1/R. ltoreq.20,

further preferably 2. ltoreq. D1/R. ltoreq.16.

5. The apparatus according to item 1, wherein the cross-basal shape of the sample placement zone further satisfies the following condition: the shape of the sample placing area on one side in the sample feeding direction of the sample to be detected is an arc which is basically outwards convex, and no obvious depression is formed on the arc.

6. The apparatus according to item 5, wherein the cross-sectional shape of the sample placement area further satisfies the following condition:

the sample placing areas are basically symmetrical along the direction vertical to the sample feeding direction of the sample to be detected.

7. The device according to any one of items 1 to 6, wherein the conduit is a microchannel having an inner diameter of 10 micrometers to 5 mm, preferably 50 micrometers to 4 mm, more preferably 100 micrometers to 3 mm, more preferably 200 micrometers to 2 mm, and more preferably 300 micrometers to 1 mm.

8. The device of any one of claims 1 to 7, wherein the sample placement section, the pulse module, the detection module, and the tubing are in fluid communication.

9. The apparatus of item 8, wherein,

in the case where the volume of the regulation region is Vo and the volume of the sample placement region is Vs,

when the pulse pressure applied by the pulse module is more than 100Pa and less than or equal to 1000Pa, the range of Vo/Vs satisfies the following conditions: 1001 is more than or equal to Vo/Vs, more preferably 500 is more than or equal to Vo/Vs, more preferably 100 is more than or equal to Vo/Vs, more preferably 50 is more than or equal to Vo/Vs, more preferably 10 is more than or equal to Vo/Vs;

when the pulse pressure exerted by the pulse module is more than 1000Pa and less than or equal to 10KPa, the range of Vo/Vs satisfies the following conditions: 101 is more than or equal to Vo/Vs, more preferably 50 is more than or equal to Vo/Vs, more preferably 10 is more than or equal to Vo/Vs, more preferably 5 is more than or equal to Vo/Vs, and more preferably 2 is more than or equal to Vo/Vs;

when the pulse pressure exerted by the pulse module is more than 10KPa and less than or equal to 100KPa, the range of Vo/Vs satisfies the following conditions: 11 is not less than Vo/Vs, more preferably not less than 5 Vo/Vs, more preferably not less than 2 Vo/Vs, more preferably not less than 1 Vo/Vs, more preferably not less than 0.8 Vo/Vs;

when the pulse module applies pulse pressure more than 100KPa, the range of Vo/Vs satisfies: 2. gtoreq.vo/Vs, more preferably 1. gtoreq.vo/Vs, further preferably 0.8. gtoreq.vo/Vs, further preferably 0.6. gtoreq.vo/Vs, further preferably 0.4. gtoreq.vo/Vs.

10. The device of any one of claims 1 to 9, wherein the conditioning zone is provided by a communication conduit between the detection module and the sample placement zone.

11. The device of any one of items 1 to 9, wherein the conditioning volume is provided by a separately provided area between the detection module and the sample placement area.

12. The device according to any one of items 1 to 11, wherein the liquid sample is a blood sample, a protein liquid sample, or other liquid sample that undergoes a phase change due to high molecular polymerization, or a substance that undergoes a phase change with a change in temperature.

13. A method for detecting a phase change in a liquid using an apparatus comprising a sample placement zone, a pulse module, a conditioning zone, a detection module, and a conduit, comprising the steps of:

the liquid sample to be detected is fed into the sample placing area through a pipeline in a mode that the liquid sample is wrapped by the medium,

providing pulsed pressure to the liquid sample through the pulse module, an

The detection module is used for detecting the change of the pressure which is applied to the sample to be detected by the detection pulse module and transmitted to one side of the detection module through the sample along with the time, and the change of the pressure along with the time is taken as a signal to be output,

the pulse module and the detection module are arranged on two sides of the sample placing area, and the adjusting area is located between the detection module and the sample placing area.

14. The method of claim 13, wherein during the performing the test, the conduit is filled with a medium selected from oil or gas, and the conditioning zone is filled with a gas, preferably the oil is mineral oil and the gas is air.

15. The method of item 13, which is used in any item 1 ~ 12 of the device for detection.

As described above, the apparatus for detecting a phase change of a liquid of the present invention has a simple structure, and can detect a coagulation time and a coagulation state of a liquid (e.g., blood) in a minute amount of a liquid sample (e.g., a blood sample amount). In addition, when the detection device of the present invention is used, the liquid sample (e.g., blood sample) is wrapped by the medium, so that the liquid sample (e.g., blood sample) is not unnecessarily disturbed by the outside during the detection process, and the whole process of the phase change of the liquid sample, such as the time when blood starts to coagulate after procoagulant drugs and factors are added and the strength during coagulation (the strength of thrombus), can be accurately detected.

In addition, the device for detecting the phase change of the liquid can detect the process of solidifying the sample from the liquid to the solid, and can further detect the process of melting the solidified solid to the liquid again. Meanwhile, as can be understood by those skilled in the art, the process from liquid to solid to liquid can be repeated for a plurality of times, and the whole process of repeating for a plurality of times can be detected by the device of the invention.

The above description is only an overview of the technical solutions of the present invention, and in order to make the technical means of the present invention more clearly apparent, and to make the implementation of the content of the description possible for those skilled in the art, and to make the above and other objects, features and advantages of the present invention more obvious, the following description is given by way of example of the specific embodiments of the present invention.

Drawings

Various other advantages and benefits of the present invention will become apparent to those of ordinary skill in the art upon reading the following detailed description of the preferred embodiments. The drawings are only for purposes of illustrating the preferred embodiments and are not to be construed as limiting the invention. It is obvious that the drawings described below are only some embodiments of the invention, and that for a person skilled in the art, other drawings can be derived from them without inventive effort. Also, like parts are designated by like reference numerals throughout the drawings.

FIG. 1 is a schematic view of one embodiment of the detection device of the present invention.

FIG. 2 is a schematic view of another embodiment of the test device of the present invention.

FIGS. 3(a) to (c) are schematic views of a sample-placing region and a cross section thereof of the detecting unit of the present invention.

FIG. 4 shows an example of the results of the test with a score of 5.

FIG. 5 shows an example of the test results with a score of 6.

FIG. 6 shows an example of the test results with a score of 7.

FIG. 7 shows an example of the results of the test with a score of 8.

FIG. 8 shows an example of the results of the test with a score of 9.

Fig. 9 cross-sectional shape of the sample-placing region employed in comparative example 1 and the results of the test scores.

Fig. 10 cross-sectional shape of the sample-placing region employed in comparative example 2 and the results of the test scores.

FIG. 11 is a graph showing the results of the detection in example 10.

FIG. 12 is a schematic view of yet another embodiment of the test device of the present invention.

FIG. 13 is a schematic view of a variant 1 of yet another embodiment of the detection device according to the invention.

FIG. 14 is a schematic view of a variant 2 of yet another embodiment of the detection device according to the invention.

FIG. 15 is a schematic view of a variant 2 of still another embodiment of the detection device of the present invention, in which detection is carried out in the presence of an oily medium.

Fig. 16 is a graph showing the detection results of embodiment 11, wherein Vo/Vs ═ 0 indicates the case where no adjustment region exists.

FIG. 17 is a schematic view of a variant 2 of yet another embodiment of the detection device of the invention, in the case of detection using air as medium.

FIG. 18 is a graph showing the results of detection in example 12.

Detailed Description

Specific embodiments of the present invention will be described in more detail below with reference to the accompanying drawings. While specific embodiments of the invention are shown in the drawings, it should be understood that the invention may be embodied in various forms and should not be construed as limited to the embodiments set forth herein. Rather, these embodiments are provided so that this disclosure will be thorough and complete, and will fully convey the scope of the invention to those skilled in the art.

< apparatus for detecting phase Change of liquid >

The device for detecting the phase change of the liquid comprises: the sample placing area is used for placing a liquid sample to be detected and completing the phase change process of the liquid in the sample placing area so as to detect the liquid sample; the pulse module is used for applying pulse pressure to the sample to be detected; the detection module is used for detecting the change of the pressure in the pipeline on one side of the detection module; a conduit having an inlet and an outlet for ingress and egress of a sample; the pulse module and the detection module are arranged on two sides of the sample placing area, and the detection of the pressure change in the pipeline on one side of the detection module means that the detection module detects the change of the pressure, which is applied to a sample to be detected by the pulse module and transmitted to one side of the detection module through the sample, along with the time, and outputs the change of the pressure along with the time as a signal.

In addition, the device for detecting liquid coagulation of the present invention may further include a sample injection module, and the sample injection module is not particularly limited as long as it is a module capable of realizing sample injection in a form in which the medium includes a liquid sample.

Fig. 1 and 2 each show a schematic view of an embodiment of the detection device according to the invention. It can be seen that, in the detecting device of the present invention, the pulse module and the detecting module need to be disposed at two sides of the sample placing region, two representative manners are shown in fig. 1 and fig. 2, respectively, but it can be understood by those skilled in the art that the pulse module and the detecting module need not be disposed at an angle of 180 degrees with respect to the sample placing region, as long as one is disposed at one side of the sample placing region and one is disposed at the other side of the sample placing region, that is, the pulse module is used to apply a pulse pressure to the sample to be detected, and the detecting module is used to detect the pressure that can be transmitted to the pipe at the other side of the sample after the pulse pressure is absorbed by the sample. Therefore, as can be understood by those skilled in the art, as the liquid sample to be measured changes from liquid to solid or from solid to liquid, the value of the pressure absorbed by the sample changes, and thus the pressure in the pipeline on the other side can be continuously changed, and the device of the present invention can characterize the change of the liquid sample to be measured from liquid to solid by detecting the change with time.

In a preferred mode, the pulse module and the detection module are positioned at 180 degrees ± 20 degrees, preferably at 180 degrees ± 10 degrees, and more preferably at 180 degrees ± 5 degrees with respect to the sample placement region.

The pulse module used in the device of the present invention applies a pulse pressure to the sample to be detected, so any means commonly used in the art for providing a pulse pressure can be used, such as a pulse pump, a plunger pump, a syringe pump, or a medium indirectly pushed with a pulse gas pressure to provide a pulse pressure. The pulsed pressure applied by the pulse module to the liquid sample to be detected during the detection process is a pulsed pressure with a substantially constant peak value of each applied pressure, for example, a certain pressure is applied at intervals, wherein the intervals can be 1-60 seconds, preferably 2-40 seconds, and further preferably 3-20 seconds; the applied pressure may be in the range of 0.1 to 50KPa, preferably 0.4 to 24KPa, more preferably 0.8 to 16KPa, and still more preferably 1.2 to 12 KPa.

The peak value of the pressure of the pulse pressure applied to the module to be detected by the pulse module is basically unchanged. The peak value is substantially constant, meaning that the pressure output of the set pulsed pressure remains constant, but that variations in the output are allowed within the tolerance of the instrument, typically to an output pressure value of ± 1KPa, preferably ± 0.5KPa, preferably ± 0.1 KPa.

The test device of the present invention should be filled with a medium throughout the device prior to use, and filling the medium throughout the device means filling the conduit and the sample placement area with the medium. When the detection device is used, a liquid sample to be detected is fed into the pipeline in a mode that the liquid sample is wrapped by the medium, and enters the sample placing area through the pipeline. And after the sample introduction is finished, opening the pulse module, and applying pulse pressure with the basically constant pressure peak value to the liquid sample to be detected. As the liquid sample gradually solidifies from a liquid state to a solid state in the sample application region, the pressure in the conduit leading to the detection block side, which is typically the pressure in the conduit at the detection block side outside the sample application region, can be detected over time by the detection means in the apparatus. Based on the output pressure signal, the detection module registers the change in pressure to reflect the entire state from liquid to coagulation.

The peak value of the gas pressure applied by the pulse module is substantially constant, so that the peak height of the pulse is substantially constant when no liquid sample is present in the device, and the peak height is the background value detected by the detection module. When the liquid sample is subjected to phase change from a liquid phase to a solid phase and then is subjected to phase change from the solid phase to the liquid phase, the transmission of pressure is influenced, so that the amplitude of a signal received by the detection module is changed, the amplitude change quantity can be used for describing the phase change intensity, the phase change intensity value can be obtained by subtracting the detection value from the background value, the phase change intensity value changing along with time is output, and a detection curve can be obtained, and the curve can represent the phase change process of the sample.

It will be understood by those skilled in the art that, since the liquid sample gradually changes from liquid to solid, the pressure pulse amplitude detected by the detection module gradually decreases, and at this time, the process of gradually decreasing the pressure pulse amplitude detected by the detection module can be recorded, and the directly measured gradually decreasing pressure pulse amplitude is subtracted from the substantially constant pulse pressure peak value, so as to obtain a gradually increasing pressure curve reflecting the solidification strength of the sample as a detection result, and various detected results can be given, for example, with reference to fig. 4 to 10. Meanwhile, if the phase transition process from solid to liquid is detected, a person skilled in the art can understand that the effect of blocking the pulse pressure by the sample in the solid state to the liquid state is weakened, so that the pulse amplitude detected by the phase transition generation detection module is continuously enhanced, and the directly measured gradually-increased pressure pulse amplitude is subtracted from the basically-constant pulse pressure peak value, so that a gradually-decreased pressure curve reflecting the liquefaction degree of the sample is obtained and is used as the detection result.

The detection module used in the device of the present invention is not limited as long as it can detect a change in pressure in the conduit, as described above, and may be any pressure sensor that can be used in the field of microfluidics. Such as micro air pressure sensors, micro hydraulic pressure sensors.

The sample placing area in the device is used for placing a liquid sample to be detected and completing the phase change process of the liquid in the sample placing area for detection. The shape of the sample-placing region is generally not limited as long as this process can be achieved. It is preferable that the cross-sectional shape of the sample-placing region of the present invention satisfies some defined conditions.

In addition, in the case of using the device of the present invention, the sample is introduced in the form of a liquid, but after the sample is introduced into the sample-placing region, the solidification of the sample from the liquid into a solid or the melting of the sample from the solid into the liquid again can be detected. Similarly, it can be known that the phase change process can be repeated multiple times, and multiple phase change processes can be detected.

Figure 3 gives a schematic representation of a cross-section of the sample placement area. FIGS. 3(a) and (b) show schematic perspective views of the apparatus of the present invention, the rectangular parallelepipeds on both sides can represent the pulse module and the detection module, respectively, and the ellipsoid or cylinder in the middle schematically represents the sample placement area. It will be understood by those skilled in the art that the shapes shown in fig. 3(a) and (b) are merely illustrative of the sample-placement region and a schematic of its cross-section, and are not intended to limit the shape of the sample-placement region. Further, fig. 3(a) and (b) show two different three-dimensional shapes, which are also exemplary, and any three-dimensional shape may be employed as long as it satisfies the shape of its cross section.

The cross section of the sample-placing region means a section obtained by cutting the center of the sample-placing region in the direction of flow of the sample with a plane of a rectangle shown in fig. 3(a) or (b), and the hatched portion indicated by oblique lines in fig. 3(a) or (b) is referred to herein as the cross section of the sample-placing region. Fig. 3(c) is a schematic plan view showing the entire cross section after cutting, in which the cross section of the sample-placing region is hatched with oblique lines. In FIG. 3, the dotted line indicates the sample introduction direction.

In the device of the present invention, it is preferable that the cross-sectional shape of the sample-placing region satisfies the following condition: the maximum width of the sample placing area in the sample feeding direction of the sample to be detected is D1 (shown in FIG. 3 (c)), the maximum width of the sample placing area in the direction perpendicular to the sample feeding direction of the sample to be detected is D2 (shown in FIG. 3 (c)), and D1 is not less than D2; the sample placing area is basically symmetrical along the sample introduction direction of the sample to be detected; and the shape of one side of the sample placing area along the sample feeding direction of the sample to be measured is an arc shape which is basically convex outwards.

In the above conditions, the condition that the sample placement area is substantially symmetrical along the sample introduction direction of the sample to be measured means that, regarding the cross section of the sample placement area, the areas of the upper part and the lower part are substantially the same and the shape is substantially symmetrical along the dotted line in the schematic diagram of fig. 3(b) as the center line. For example, the area of the portion on the imaginary line and the area of the portion under the imaginary line are different by 20%, more preferably by 15%, further preferably by 10%, further preferably by 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, and 1%.

The shape of one side of the sample placement area in the sample introduction direction of the sample to be measured is an arc shape that is substantially convex outward. An arc is the shape of a portion of a circle or ellipse. The substantially outwardly convex arc means a shape in which the arc protrudes outward from the dotted line in fig. 3, and the arc means an arc which looks like a part of a circle or an ellipse as a whole, and is not required to be completely smooth.

In the device of the present invention, it is further preferable that the cross-sectional shape of the sample-placing region satisfies the following condition: 1 < D1/D2. ltoreq.8, preferably 1 < D1/D2. ltoreq.7, more preferably 1 < D1/D2. ltoreq.6, still more preferably 1 < D1/D2. ltoreq.5. That is, the cross section of the sample placement section of the present invention preferably has a shape having a major axis and a minor axis, wherein the direction of introduction is the major axis and the direction disposed in line with the direction of introduction is the minor axis.

In the device of the present invention, it is further preferable that the cross-sectional shape of the sample-placing region satisfies the following condition: when the inner diameter of the pipe for the inlet and outlet through which the sample is introduced and discharged is R, 2. ltoreq. D1/R. ltoreq.24, preferably 2. ltoreq. D1/R. ltoreq.20, and more preferably 2. ltoreq. D1/R. ltoreq.16 are satisfied. That is, the volume of the sample-placing region of the present invention and the size of the tube preferably satisfy the above-mentioned relationship.

In the device of the present invention, it is further preferable that the shape of the transverse basal plane of the sample placement area further satisfies the following condition: the shape of the sample placing area on one side in the sample feeding direction of the sample to be detected is an arc which is basically outwards convex, and no obvious depression is formed on the arc. By recessed is meant herein, for example, the occurrence of a portion such as a valley in the arc, such as the cross-sectional shape of the sample placement area in examples 6-8 described below. Examples 6 and 8 had one distinct depression in the arc on one side in the direction of sample introduction, and example 7 had two distinct depressions in the arc on one side in the direction of sample introduction.

In the device of the present invention, it is further preferable that the cross-sectional shape of the sample-placing region also satisfies the following condition: the sample placing area is basically symmetrical along the direction vertical to the sample feeding direction of the sample to be detected.

In another embodiment of the present invention, an apparatus for detecting a phase change of a liquid comprises: the sample placing area is used for placing a liquid sample to be detected and completing the phase change process of the liquid in the sample placing area so as to detect the liquid sample; the pulse module is used for applying pulse pressure to the sample to be detected; the detection module is used for detecting the change of the pressure in the pipeline on one side of the detection module; the adjusting area is used for adjusting the intensity of a signal fed back by the sample to be detected and detected by the detection module, wherein the intensity of the detection signal is improved when the volume of the adjusting area is increased; when the volume of the adjusting area is reduced, the intensity of the detection signal is reduced; a conduit having an inlet and an outlet for ingress and egress of a sample; the pulse module and the detection module are arranged on two sides of the sample placing area, the detection of the change of the pressure in the pipeline on one side of the detection module means that the detection module detects the change of the pressure, applied to a sample to be detected by the pulse module and transmitted to one side of the detection module through the sample, along with the time, and outputs the change of the pressure along with the time as a signal, and the adjusting area is positioned between the detection module and the sample placing area.

In the present invention, it is possible for the regulating region to be formed of any structure as long as its volume is larger than 0. Fig. 12 shows a schematic view of a device for detecting a phase change of a liquid comprising a conditioning region, and it will be understood by those skilled in the art that fig. 12 only schematically shows the position of the conditioning region, and there is no limitation on the shape and size thereof.

In another specific embodiment, the conditioning zone is provided by a communication channel between the detection module and the sample placement zone, as shown in FIG. 13. In addition, although the conditioning region is formed by expanding the tube in FIG. 13, it will be understood by those skilled in the art that the space between the sample and the detection module can be used as the conditioning region.

In another specific embodiment, the modulated volume region is provided by a separately disposed region between the detection module and the sample placement region, as shown in FIG. 14. It will also be appreciated by those skilled in the art that FIG. 14 is also merely a schematic representation of the conditioning zone as a separately disposed region and that FIG. 14 is not intended to limit the shape and size of the conditioning zone.

Further, in the present invention, in the case where the volume of the regulation region is Vo and the volume of the sample placement region is Vs, when the pulsed pressure applied by the pulse module is greater than 100Pa and equal to or less than 1000Pa, the range of Vo/Vs satisfies: 1001 is more than or equal to Vo/Vs, more preferably 500 is more than or equal to Vo/Vs, more preferably 100 is more than or equal to Vo/Vs, more preferably 50 is more than or equal to Vo/Vs, more preferably 10 is more than or equal to Vo/Vs; when the pulse pressure exerted by the pulse module is more than 1000Pa and less than or equal to 10KPa, the range of Vo/Vs satisfies the following conditions: 101 is more than or equal to Vo/Vs, more preferably 50 is more than or equal to Vo/Vs, more preferably 10 is more than or equal to Vo/Vs, more preferably 5 is more than or equal to Vo/Vs, and more preferably 2 is more than or equal to Vo/Vs; when the pulse pressure exerted by the pulse module is more than 10KPa and less than or equal to 100KPa, the range of Vo/Vs satisfies the following conditions: 11 is not less than Vo/Vs, more preferably not less than 5 Vo/Vs, more preferably not less than 2 Vo/Vs, more preferably not less than 1 Vo/Vs, more preferably not less than 0.8 Vo/Vs; when the pulse module applies pulse pressure more than 100KPa, the range of Vo/Vs satisfies: 2. gtoreq.vo/Vs, more preferably 1. gtoreq.vo/Vs, further preferably 0.8. gtoreq.vo/Vs, further preferably 0.6. gtoreq.vo/Vs, further preferably 0.4. gtoreq.vo/Vs. By controlling the size of the adjusting area, the strength of a signal fed back by a sample to be detected and detected by the detection module can be effectively adjusted, so that the phase change process of the sample to be detected can be better detected. By appropriately increasing the signal strength by the conditioning region, a higher resolution of detection can be provided, the overall phase change process is described more clearly, and even some details of the change may be shown that are masked at lower signal strengths than would be possible without the conditioning region.

The cross-sectional shapes that can be used and cannot be used are shown in table 1 of the inventive examples and comparative examples. It will be fully understood by those skilled in the art that only schematic cross-sections are shown in table 1, and shapes other than those shown in table 1 may be used or may be more preferably used as long as the above-described limitations of the present invention are satisfied.

In the device of the present invention, the tube is a microchannel, and the inner diameter thereof is 10 μm to 5 mm, preferably 50 μm to 4 mm, more preferably 100 μm to 3 mm, further preferably 200 μm to 2 mm, and further preferably 300 μm to 1 mm.

In the present invention, the material of the conduit is not limited, and any material may be used as long as it can wrap the liquid sample with the medium by the operation of the present invention. Examples thereof include: polymer materials such as Polytetrafluoroethylene (PTFE), polymethyl methacrylate (PMMA), polypropylene (PP), etc., glass, and metal materials such as stainless steel, titanium alloy, copper, platinum, gold, etc.

In the present invention, a certain amount of the liquid sample to be measured is required to be fed to the sample placement area, and the certain amount of the liquid sample to be measured refers to the amount of the liquid sample to be measured in the present invention, and the certain amount is not specifically limited, and the skilled person knows how to select an appropriate amount according to the subsequent processing or detection of the sample, so as to measure the coagulation process of the liquid sample, such as: 0.1-200 microliter, 0.5-150 microliter, 1-100 microliter, 2-80 microliter, 3-60 microliter and the like, and can be specifically 200 microliter, 150 microliter, 100 microliter, 80 microliter, 60 microliter, 40 microliter, 20 microliter, 10 microliter, 8 microliter, 6 microliter, 4 microliter, 2 microliter, 1 microliter, 0.1 microliter and the like.

In the present invention, as described above, the liquid sample is a blood sample or a polymer solution such as a protein solution, a gelatin solution, a protein curd, a polymer material solution, or the like.

The medium used in the present invention is generally a lipophilic medium, and various kinds of oils generally used in the art can be used. Such as mineral oil, low temperature paraffin, vegetable oil.

< method for detecting phase transition of liquid >

The invention also provides a method for detecting the phase change of liquid, which uses a device comprising a sample placing area, a pulse module, a detection module and a pipeline for detection and comprises the following steps: the detection device comprises a detection module, a pulse module, a detection module and a signal output module, wherein the detection module is used for detecting the change of the pressure applied to the sample to be detected by the detection module and transmitted to one side of the detection module through the sample along with the time, and the change of the pressure along with the time is used as the signal output, wherein the pulse module and the detection module are arranged at two sides of the sample placement area.

The invention also provides a method for detecting the phase change of liquid, which uses a device comprising a sample placing area, a pulse module, a regulating area, a detection module and a pipeline for detection and comprises the following steps: the method comprises the steps of feeding a liquid sample to be detected to a sample placing area through a pipeline in a medium-wrapped liquid sample mode, providing pulse pressure for the liquid sample through a pulse module, detecting the change of the pressure, applied to the sample to be detected by the pulse module and transmitted to one side of the detection module through the sample, of the detection module along with time by utilizing the detection module, and outputting the change of the pressure along with time as a signal, wherein the pulse module and the detection module are arranged on two sides of the sample placing area, and the adjusting area is located between the detection module and the sample placing area. The regulatory region is as described above.

In the present invention, during the detection, the duct is filled with a medium selected from oil or gas, the conditioning zone is filled with a gas, preferably the oil is mineral oil and the gas is air.

The apparatus used in the process of the invention is the apparatus of the invention described above.

In addition, the method of the present invention further comprises: the entire detection system is filled with medium prior to detection.

As described above, the apparatus for detecting a phase change of a liquid of the present invention has a simple structure, and can detect a coagulation time and a coagulation state of a liquid (e.g., blood) in a minute amount of a liquid sample (e.g., a blood sample amount). In addition, when the detection device of the present invention is used, the liquid sample (e.g., blood sample) is wrapped by the medium, so that the liquid sample (e.g., blood sample) is ensured not to be interfered by outside unnecessarily during the detection process, and the whole process of the coagulation of the liquid sample can be accurately detected, for example, the time for blood to start to coagulate becomes short and the strength during coagulation (the strength of thrombus) becomes large after procoagulant drugs and factors are added.

In addition, in the case of a blood sample, the sample may liquefy within a certain period of time after coagulation, and this change indicates that fibrinolysis of the coagulated blood occurs, which is medically referred to as hyperfibrinolysis; patients with hyperfibrinolysis have a high risk of internal bleeding, and if the patients need to be operated, medical measures must be taken to improve the coagulation capacity of the patients, otherwise the patients can bleed a lot during or after the operation, and even become life-threatening. The device can detect the processes and realize the detection of the hyperfibrinolysis process.

In addition, since the apparatus of the present invention employs a given sample introduction module as described below, different liquid samples can be continuously introduced to achieve continuous, one-time processing of a large number of different liquid samples. The liquid samples fed each time can be properly isolated from each other and are not mutually polluted and influenced, so that continuous feeding of various liquid samples and effective detection can be realized.

Examples

The construction of the detection device used in the following examples is substantially in the manner of fig. 2.

The pulse module consists of a constant pressure air source and a time control electromagnetic valve. Adopting a Kammer brand KLP01 type diaphragm pump purchased from Kachuan er fluid science and technology (Shanghai) Limited company, a Song brand DP-101 type air pressure sensor switch module purchased from Shenzhen Chun Toyobo instrument Limited company and a 2L capacity stainless steel air storage tank to form a set of constant pressure air source, wherein the air source is positive pressure; the time control solenoid valve is formed by adopting a Sono Tian Gong brand TM-06 two-position three-way high-frequency solenoid valve purchased from Ningzhen Shen Sheng electronics Limited and a DTM01 type time relay module purchased from Shenzhen Sono industry automatic control equipment Limited.

Wherein, the normally closed port of the electromagnetic valve is connected with a constant pressure air source, the normally open port is connected with a pulse pipeline communicated with the fluid in the detection area, the pipeline is filled with a medium, and the other port of the electromagnetic valve is communicated with the outside air. When the electromagnetic valve does not act, the pulse channel is communicated with the outside air through the electromagnetic valve, and the pressure detected by the detection module is 0 Pa; when the electromagnetic valve acts, the electromagnetic valve is communicated with the constant-pressure air source, and the pressure detected by the detection module is a certain specific pressure value which is greater than zero and less than or equal to the pressure of the constant-pressure air source. The action time and the interval time of the electromagnetic valve are controlled by a time relay module.

Meanwhile, the negative pressure constant pressure air source is manufactured by adopting the same parts and method for manufacturing the constant pressure air source. The method is used for sample introduction operation of a sample to be detected.

The medium used in the examples described below was mineral oil and the source pressure was controlled at 12 KPa.

A pipe having an inner diameter of 0.6mm × 0.6mm, the pipe being made of polymethyl methacrylate (PMMA);

the sample-placing region is made of polymethyl methacrylate (PMMA) material, and in the examples, the three-dimensional structure of the sample-placing region is a cylindrical structure, as shown in fig. 3(b), and the cross-sectional shapes and parameters specifically adopted in the respective examples are listed in table 1 below.

The detection module uses a miniature gas-liquid universal pressure sensor (XGZP 6847 pressure sensor module purchased from WU lake core sensory sensor technology, Inc.) to output a 0-5V voltage signal with a measuring range of 0-20 KPa.

The following materials were used in examples 1 to 9 and comparative examples 1 to 4:

sheep plasma (for heparin sodium titer detection) was purchased from litsea cubeba to Ming Biochemical auxiliary factory, stored at-18 ℃ and thawed at 4 ℃ before the experiment and activated at 37 ℃ for 1 hour.

The calcium chloride solution purchased from the company Hissemcang Biotechnology (Wuxi) Ltd has a calcium ion concentration of 0.02mol/L, is prepared at the above concentration, is stored at a low temperature of 4 ℃ and is preheated to room temperature when used. The calcium ion is used for promoting coagulation reaction of sheep plasma.

After the experimental material is prepared, taking the activated goat plasma, adding calcium chloride, and timing (taking 0 second at this time), wherein the sample is positioned in a centrifugal tube; the sample was injected into the sample placement area and the test was started at the 100 second time point.

Before sample introduction, the pulse port is closed, the sample outlet pipeline is connected with a negative pressure constant pressure air source through a valve, and the system is filled with a medium. When the sample is introduced, a pipeline connected with a sample inlet extends into a centrifugal tube filled with a sample, a valve of a sample outlet pipeline connected with a negative pressure constant pressure air source is opened, the sample is sucked to a sample placing area through the inlet, the sample placing area is full of the sample, the valve of the sample outlet pipeline connected with the negative pressure constant pressure air source is closed, the centrifugal tube filled with the sample to be detected is removed, the sample inlet is closed, and the sample introduction is completed.

And after the sample introduction is finished, the pulse port is opened, and the pulse channel is communicated with the pulse module through fluid. And starting the time relay module to enable the electromagnetic valve of the pulse module to intermittently act, so that pulse pressure is applied to the sample to be detected, and detection is started. The pulse period is 5S interval 5S of applied pressure, the pressure of the positive pressure constant pressure air source of the pulse module is 12KPa, and the pressure of the negative pressure constant pressure air source is negative 6 KPa. The detection time is 12-20 min.

Fig. 4 to 8 show examples of scores of different test results when the test device of the present invention is used for testing, and examples 1 to 9 below are scored according to the examples of fig. 4 to 8. As can be seen from fig. 4, the example with the score of 5 can basically describe the complete phase transformation process, but the detailed description of the phase transformation process is limited, and the main characteristic of the phase transformation occurrence is not completely reflected. As can be seen from fig. 5, the example with a score of 6 may describe a complete phase transformation process, and although the detailed description of the phase transformation process is not accurate enough, the main characteristics of the phase transformation occurring may be substantially reflected. As can be seen from fig. 6, the 7-point example can describe the complete phase transformation process, and although the local detailed description of the phase transformation process is not accurate enough, the main characteristics of the phase transformation occurrence can be reflected. As can be seen from fig. 7, the 8-point example can accurately and completely describe the entire phase transformation process, and only a small data deviation exists locally, but the accuracy of the description of the phase transformation process is not substantially affected. As can be seen from fig. 8, the example of the score of 9 can accurately and completely describe the whole phase transformation process, and only the individual data points have small fluctuation, but do not affect the accuracy of the description of the phase transformation process.

Example 10

Using the same apparatus as in examples 1 to 9, the sample-placing section having a cross section exactly the same as that of example 3 was placed in an incubator, and was in fluid communication with a pulse module and a detection module placed outside the incubator through a polytetrafluoroethylene hose having an inner diameter of 0.6 mm. A blue leopard brand LRH-150 incubator purchased from Shanghai-constant technologies, Inc. was used. As a sample, margarine for baking purchased from Tian Mei Hua Dai food Co., Ltd, which is called Henhaote, was used, and the sample was in a soft solid state at 28 to 34 ℃ and in a liquid state at 34 ℃ or higher.

The sample placement area of the device and the sample were preheated at 40 ℃ for 20 minutes before the experiment, and the sample was completely melted into a liquid state. Using the same sample introduction method as in examples 1 to 9, a liquid sample was introduced into the sample placement area, and then detection was started with a detection start time of 0 second. After 1 minute from the start of the detection, the temperature of the constant temperature incubator was set to 25 ℃ and the liquid sample started to gradually become solid as the temperature decreased; after the test had proceeded for 12 minutes, the sample had solidified sufficiently, and the incubator temperature was again set to 40 ℃ and the solid sample gradually returned to the liquid state again as the temperature increased. The detection module completely records the change process of the sample from the liquid state to the solid state and then from the solid state to the liquid state, and the specific result is shown in fig. 11.

In comparative examples 1 and 2, the shape of the transverse base surface of the sample-placing section was such that the length in the sample-introducing direction was smaller than the length in the direction perpendicular to the sample-introducing direction. When the shapes were two kinds of shapes having a sample direction ratio of 1:1.5 in a direction perpendicular to the sample direction, the results showed only a limited response to the occurrence of phase transition and the process was not described, and fig. 9 shows the cross-sectional shapes and results of the sample-placing regions used in comparative example 1 and comparative example 2. As can be seen from FIG. 9, the satisfactory detection curve could not be obtained in the sample-placing region not satisfying the above-mentioned condition D1. gtoreq.D 2.

In comparative examples 3 and 4, if the shape of the transverse base surface of the sample-placing section is asymmetrical in the sample-feeding direction. As shown in fig. 10, when the shape is a first asymmetric shape, the result is a limited response to phase change and the process is not fully described; and when the shape is the second asymmetric pattern, the result can basically describe the occurrence of phase change, but cannot describe the complete process of the phase change, and the detection is unstable in the later period. It can be seen from fig. 10 that a satisfactory detection curve cannot be obtained for a sample placement area that does not satisfy the conditions that the sample placement area is substantially symmetrical in the direction of introduction of the sample to be measured and the shape of one side of the sample placement area in the direction of introduction of the sample to be measured is an arc that is substantially convex outward.

On the other hand, the average value of the results of three tests in examples 1 to 9 is at least 5 points, and the purpose of detecting the liquid phase change process can be achieved.

Table 1 summarizes the cross-sectional shapes of the sample-placing areas of examples 1 to 9 and the results of scoring three test results according to the scoring examples of fig. 4 to 8.

Example 11

The device was constructed in the same manner as in example 3 above, except that there was a conditioning zone between the detection module and the sample placement zone. The sample placement area was elliptical as in example 3, and the adjustment area was an independent area communicating with the conduit between the detection module and the sample placement area through a thin conduit; as shown in fig. 15. Before detection, the detection module is taken down, all pipelines are filled with mineral oil, air with a fixed volume can be sealed in the adjusting area, and then the detection module is connected, so that the integral structure of the detection module, the adjusting area, the sample placing area and the pulse module in the embodiment is formed.

After the device was assembled, the sheep plasma samples were tested for the sample injection and coagulation process in a manner complete to example 1, wherein figure 15 shows a schematic diagram of the testing process, wherein the dark grey areas represent the samples, the hatched areas represent the areas filled with mineral oil, and the conditioned area is air.

In this embodiment, the pressure of the positive pressure constant pressure air source connected to the pulse module is 12KPa, the pressure of the negative pressure constant pressure air source for sample injection is controlled to be negative 6KPa, and the detection time is 12-20 min.

The detection results of example 11 are shown in FIG. 16, and it can be seen that the signals of examples 11-1, 11-2, and 11-3 with the adjustment regions are stronger than the signals of example 3 without the adjustment regions, and as the adjustment regions are increased, Vo/Vs is increased, the signals are gradually enhanced, and the resolution of the detection process is correspondingly improved after the signals are enhanced, so the detection effect is further improved.

Example 12

The apparatus was constructed in the same manner as in example 11 above, with the conditioning zone being an independent area communicating with the conduit between the detection module and the sample placement zone via a thin conduit; before detection, the whole device is filled with air. After the sample introduction is finished, the volume occupied by the air sealed in the sample and the detection module is the volume of the regulation area, and the sealed area plays the role of the regulation area.

The difference from example 11 is only that in the apparatus for detection, no mineral oil was injected, but air was used as a medium, and fig. 17 shows a schematic view during detection, and a dark gray area indicates a sample, except that other spaces in the apparatus were filled with air. As shown by the dotted line in the figure, a part of the area enclosed by the dotted line is the gas volume contained by the conduit, communication structure between the detection module and the sample; the other part is to supplement the added volume between the detection module and the sample, which volume in its entirety constitutes the volume of the conditioning zone.

In example 12, the detection zone uses the same elliptical configuration as in example 11 and the other parameters are as follows:

example 12 was conducted in the same manner as in examples 1-9 above, except that the sample was changed to human whole blood and human venous blood was drawn into a citrated blood collection tube. Before detection, 1mL of blood sample is placed in a kaolin activated reagent tube, shaken up and waits for 1min, then 340 μ L of activated blood sample is added into 20 μ L of calcium chloride solution (0.2mol/L), the blood sample added with calcium chloride is injected into a sample placement area, a sample inlet is closed, and detection is started, wherein the detection result is shown in FIG. 18.

Because the source, the component and the biochemical properties of the goat plasma and the human whole blood are essentially different, the signal intensity in the detection is greatly different, and the difference is mainly caused by the sample.

From the results of fig. 18, it can be seen that Vo/Vs increases and the signal increases when the volume of the adjustment region increases, which shows that the detection effect can be improved by the adjustment region in the case of air as well.

The results of the above examples show that the time at which the liquid solidifies and the intensity of the solidification of the reaction liquid can be effectively detected using the apparatus of the present invention.

The present application is susceptible to various modifications and alternative forms, specific embodiments have been shown by way of example in the drawings and have been described in detail herein. However, the application is not intended to be limited to the particular forms disclosed. On the contrary, the intention is to cover all modifications, equivalents, and alternatives falling within the scope of the application, which is defined by the appended claims and their legal equivalents.

The numerical ranges recited in the present invention each include data for both endpoints of the numerical range, and also include each of the specific values in the numerical range, and the numerical values can be combined with the endpoints at will to form a new subrange.

Claims

1. An apparatus for detecting a phase change of a liquid, comprising:

a conduit having an inlet and an outlet for ingress and egress of a sample;

2. The apparatus of claim 1, wherein the cross-sectional shape of the sample placement area satisfies the condition:

3. The apparatus of claim 2, wherein the cross-sectional shape of the sample placement area further satisfies the condition:

1＜D1/D2≤8，

preferably 1 < D1/D2. ltoreq.7,

more preferably 1 < D1/D2. ltoreq.6,

further preferably 1 < D1/D2. ltoreq.5.

4. The apparatus of claim 3, wherein the cross-sectional shape of the sample placement area further satisfies the condition:

satisfying 2 < D1/R < 24 >,

preferably 2. ltoreq. D1/R. ltoreq.20,

further preferably 2. ltoreq. D1/R. ltoreq.16.

5. The apparatus of claim 1, wherein the sample placement area further has a transverse basal shape that satisfies the following condition: the shape of the sample placing area on one side in the sample feeding direction of the sample to be detected is an arc which is basically outwards convex, and no obvious depression is formed on the arc.

6. The apparatus of claim 5, wherein the cross-sectional shape of the sample placement area further satisfies the condition:

7. The device according to any one of claims 1 to 6, wherein the conduit is a microchannel having an inner diameter of 10 micrometers to 5 mm, preferably 50 micrometers to 4 mm, more preferably 100 micrometers to 3 mm, more preferably 200 micrometers to 2 mm, more preferably 300 micrometers to 1 mm.

8. The device of any one of claims 1 to 7, wherein the sample placement section, the pulse module, the detection module, and the conduit are in fluid communication.

9. The apparatus of claim 8, wherein,

when the pulse module applies pulse pressure more than 100KPa, the range of Vo/Vs satisfies:

2. gtoreq.vo/Vs, more preferably 1. gtoreq.vo/Vs, further preferably 0.8. gtoreq.vo/Vs, further preferably 0.6. gtoreq.vo/Vs, further preferably 0.4. gtoreq.vo/Vs.