CN116297064A - Method and device for measuring dynamic multiple parameters of particles in solution - Google Patents
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- G01N15/00—Investigating characteristics of particles; Investigating permeability, pore-volume or surface-area of porous materials
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- G—PHYSICS
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Abstract
The invention relates to the technical field of optical detection, and provides a method and a device for measuring dynamic multiple parameters of particles in a solution. The device comprises a channel for circulating the solution to be measured, a light source for emitting laser and irradiating the solution to be measured, and an optical receiver for receiving the light passing through the solution to be measured; the optical path formed by the light source is perpendicular to the channel. The invention can not only realize the measurement of bubbles in any solution, but also realize the measurement of various parameters in the culture solution containing animal cells, in particular the accurate quantitative test of the concentration of cancer cells by taking the laser with the wavelength of 620-970 nm as a light source, and has very important research significance.
Description
Technical Field
The invention relates to the technical field of optical detection, in particular to a method and a device for measuring dynamic multiple parameters of particles in a solution.
Background
The application of optical density methods in mineral suspensions has been a few decades history and has recently been used in microbiology more often, for example, as a simple and rapid indirect measurement method for estimating the biomass concentration in a liquid, with the advantages of short analysis time, simple experiment, reduced cost and no damage to the sample.
At present, the object of optical density detection is often algae cells and colonies and other microorganisms, and although the optical density can also be related to the cell concentration for measuring the cell concentration, the relationship between the optical density and the cell concentration is weakened due to complex interaction between light and suspended substances, and the interaction depends on the wavelength of a light source, the concentration of cells, the size distribution of cells and the like.
In the research of optical responses of microorganisms such as bacteria, fungi, microalgae, etc., many scholars have performed optimal wavelength selection according to the optical responses of various microorganisms to improve the accuracy of the optical density measurement method of the microorganism solution. For example, bacterial cultures (e.coli, etc.) are usually selected to be measured at around 600 nm; fungal cultures such as yeasts can be selected for measurement at a wavelength of 660 nm; the recommended absorbance measurement wavelength for the measurement of the green (chlorella) and blue-green (mat algae) algae biomass is 680nm. However, there are still few people using optical density to quantitatively measure the cell concentration of cancer cells. The tumor cells are used as experimental objects frequently used in biotechnology, have great research value and potential for novel cell therapy, and the optical density measurement method of the related cancer cells has certain industrial application prospect due to the advantages of rapidness, simplicity in operation and no marker.
In view of this, the present invention has been proposed.
Disclosure of Invention
The invention provides a method and a device for measuring dynamic multiple parameters of particles in a solution, which widen the application range of an optical density measurement method by taking laser with the wavelength of 620-970 nm as a light source, and particularly realize quantitative tests of the concentration of animal cells (such as cancer cells) in various culture solutions and tests of other parameters, such as whether a culture medium contains bubbles, a phenol red indicator and a pH value.
The invention aims to provide a method for measuring particle parameters in a solution, wherein a test object of the method can be a static solution or a dynamic solution, and by taking laser with the wavelength of 620-970 nm as a light source, not only can the measurement of bubbles in any solution be realized, but also the measurement of various parameters in a culture solution containing animal cells, in particular the accurate quantitative test of the concentration of cancer cells, and the method has very important research significance.
The second object of the present invention is to provide a device for measuring particle parameters in solution, which uses optical sensing to non-invasively measure the concentration of cells in a static or flowing state, and can build a customized cell optical density sensor to provide accurate real-time cell concentration measurement, which has strong practical value for quantitative monitoring and sustainable production of biological products.
The invention also provides a method for measuring the particle parameters in the solution, which comprises the following steps: and using laser with the wavelength of 620-970 nm as a first light source to irradiate the solution to be measured, using an optical receiver to receive the light passing through the solution to be measured, and judging whether the solution to be measured contains bubbles and/or the concentration of particles in the solution to be measured.
The solution to be tested may be either static or dynamic.
Among the lasers having a wavelength of 620 to 970nm, a laser having a wavelength of 800 to 900nm is more preferable.
According to the method for measuring the particle parameters in the solution, the optical receiver is a photoelectric sensor and outputs voltage; the judging comprises the following steps:
when the voltage is positive voltage, if the detected voltage value is larger than (V1 +V2)/2, the air bubble is contained, otherwise, the air bubble is not contained;
when the voltage is a negative voltage, if the detected voltage value is smaller than (V1 +V2)/2, the air bubble is contained, otherwise, the air bubble is not contained;
wherein V1 is a voltage value when air is detected under the same condition, and V2 is a voltage value when water is detected under the same condition.
According to the method for measuring the particle parameters in the solution provided by the invention, the solution to be measured is a static or fluid dynamic culture solution containing animal cells, the particles are the animal cells in the culture solution, and preferably, the animal cells are suspended cells.
The animal cells may be cancer cells, T cells, stem cells, etc.
According to the method for measuring the particle parameters in the solution provided by the invention, the measuring process of the animal cells in the culture solution comprises the following steps:
(1) Testing the culture solution with known animal cell concentration under the wavelength to obtain corresponding OD value, and drawing a fitting curve of the relevant concentration and the OD value;
the fitting curve in the invention can be obtained by adopting a piecewise fitting mode or can be obtained by adopting a linear regression fitting mode, and generally, the regression coefficient is required to be more than 0.985. In addition, although the fitting curve is used as a better calculation means, the method of the invention can also adopt other ways of calculating the concentration, such as an interpolation method.
The medium in the solution to be measured in the invention is the same as the medium in the standard solution adopted in the process of obtaining the fitting curve.
(2) Substituting the OD value measured by the solution to be measured under the wavelength into the fitting curve to obtain the concentration of the animal cells in the solution to be measured.
The method for measuring the particle parameters in the solution provided by the invention further comprises the following steps: irradiating the solution to be detected by using laser with the wavelength of 300-600 nm as a second light source, and judging whether the culture solution contains a phenol red indicator or not;
preferably, the second light source is a laser with a wavelength of 320-470 nm or 500-580 nm;
more preferably, a laser having a wavelength of 560.+ -.20 nm is used as the second light source.
According to the method for measuring the particle parameters in the solution provided by the invention, the measuring process comprises the following steps:
the absorbance obtained by irradiating the solution to be measured by using laser with the wavelength of 300-600 nm as a second light source is marked as A;
calculating the equivalent absorbance of the cell concentration corresponding to the solution to be detected under the wavelength used by the second light source, and marking as B;
if A/B is more than 0.2, the solution to be measured contains a phenol red indicator, otherwise, the solution to be measured does not contain the phenol red indicator.
According to the method for measuring the particle parameters in the solution provided by the invention, if the culture solution contains the phenol red indicator, the measuring method further comprises the following steps: irradiating the solution to be detected by using laser with the wavelength of 300-600 nm as a second light source, and judging the pH value of the culture solution;
preferably, the second light source is a laser with a wavelength of 320-470 nm or 500-580 nm;
more preferably, a laser having a wavelength of 560.+ -.5 nm is used as the second light source.
According to the method for measuring the particle parameters in the solution provided by the invention, the measuring process comprises the following steps:
the absorbance obtained by irradiating the solution to be measured by using laser with the wavelength of 300-600 nm as a second light source is marked as A;
calculating the equivalent absorbance of the cell concentration corresponding to the solution to be detected under the wavelength used by the second light source, and marking as B;
let a/b=c, substituting C into the absorbance standard curve of the standard pH phenol red solution to obtain the pH of the culture solution.
According to the method for measuring the particle parameters in the solution, the number of the lasers is more than 2, and the lasers are used for simultaneously measuring the concentration of animal cells in the culture solution and other parameters;
the other parameters include: whether the culture solution contains bubbles, whether the culture solution contains a phenol red indicator and/or the pH value of the culture solution.
The invention also provides a device for measuring the particle parameters in the solution, which comprises: a channel for circulating the solution to be measured, a light source for emitting laser light and irradiating the solution to be measured, and an optical receiver for receiving light passing through the solution to be measured; the optical path formed by the light source is perpendicular to the channel;
preferably, the number of the light sources is more than 2;
more preferably, the equivalent diameter of the cross section of the channel is 0.5 mm-15 mm, and the equivalent diameter variation value of the cross section in the flow direction is less than 1mm, and the transmittance of the laser passing through the channel is more than 80 percent when the laser passes through the channel; preferably 90% or more.
The device body where the channel is located is manufactured by adopting a 3D printing technology by taking transparent photosensitive resin as a raw material, and can also be made of transparent materials by adopting a machining or demolding method.
According to the method for measuring the particle dynamic multi-parameter in the solution, provided by the invention, the wavelength in a specific range is selected as the irradiation light source, so that the measurement of the particle parameters in various solutions can be realized, and the application range of an optical density method is greatly expanded.
According to the method for measuring the particle dynamic multi-parameter in the solution, provided by the invention, the contribution of a container, a medium and the like to the optical density is minimized by selecting the laser with the wavelength of 620-970 nm, and the static or flowing cell concentration can be further measured.
The dynamic multi-parameter measuring device for the particles in the solution can measure the cell concentration on line with good precision and repeatability according to the output voltage value or the absorbance value of the solution to be measured, and the compact structure of the sensor also makes the device a convenient way for estimating the cancer cell concentration.
Drawings
In order to more clearly illustrate the invention or the technical solutions of the prior art, the following description will briefly explain the drawings used in the embodiments or the description of the prior art, and it is obvious that the drawings in the following description are some embodiments of the invention, and other drawings can be obtained according to the drawings without inventive effort for a person skilled in the art.
FIG. 1 shows a standard curve obtained at six wavelengths of 410nm, 560nm, 580nm, 750nm, 850nm and 970nm with cell concentration as X axis and OD (optical density) as Y axis, wherein a is 410nm, b is 560nm, c is 580nm, d is 750nm, e is 850nm and f is 970nm.
FIG. 2 is an OD calibration curve at 850nm for K562 cells (suspension cells) and A549 cells (adherent cells), where a is K562 cells using physiological saline as medium, b is K562 cells using RPMI-1640 cell culture medium, c is A549 cells using physiological saline as medium, and d is A549 cells using DMEM cell culture medium.
Fig. 3 is a schematic structural diagram of a device for measuring particle parameters in a solution according to the present invention.
Fig. 4 is another schematic structural diagram of a device for measuring particle parameters in a solution according to the present invention.
FIG. 5 is a graph of the equivalent absorbance of HELA cells at 570nm wavelength in example 8 provided by the present invention.
FIG. 6 is a graph showing the fit curves of example 8, which are referred to in determining pH, wherein (a) is a fit curve corresponding to 3 < pH < 7, and (b) is a fit curve corresponding to 7.ltoreq.pH < 10.
Reference numerals:
1: solution inlet, 2: solution outlet, 3: a channel through which the solution to be measured flows, 4: photoelectric sensing area, 5: and a laser emission region.
Detailed Description
For the purpose of making the objects, technical solutions and advantages of the present invention more apparent, the technical solutions of the present invention will be clearly and completely described below with reference to the accompanying drawings, and it is apparent that the described embodiments are some embodiments of the present invention, not all embodiments. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.
The method and apparatus for measuring the dynamic multi-parameter of the particles in the solution according to the present invention are described below with reference to fig. 1 to 6.
The cell concentration equivalent absorbance curve of the solution to be detected containing cells under a specific wavelength is obtained by measuring the absorbance of colorless standard physiological saline containing different cell concentrations by using a spectrometer and fitting the absorbance, the excitation wavelength of the spectrometer is 200 nm-1000 nm, the cell concentration equivalent absorbance curve corresponding to any value in the wavelength range can be obtained, and if the concentration of a certain solution to be detected containing cells is determined, the equivalent absorbance corresponding to the concentration can be calculated by the cell concentration equivalent absorbance curve under the corresponding wavelength.
Example 1 measurement device for particle parameters in solution
A device for measuring particle parameters in a solution, as shown in fig. 3, comprising: a channel 3 for flowing the solution to be measured (including a solution inlet 1 and a solution outlet 2), a light source (located at a laser emitting area 5) for emitting laser light and irradiating the solution to be measured, and an optical receiver (such as a SIEMENS photodiode BPW34 located at a photoelectric sensing area 4) for receiving light passing through the solution to be measured; the equivalent diameter of the cross section of the channel is 0.5-15 mm, and the equivalent diameter variation value of the cross section in the flow direction is less than 1mm; the optical path formed by the light source is perpendicular to the channel, and the light transmittance of the channel at the laser passing position is more than 80% when the laser passes through the channel; the device body where the channel is located is manufactured by adopting a 3D printing technology by taking transparent photosensitive resin as a raw material.
Example 2 measurement device for particle parameters in solution
An optical device for measuring parameters of particles of a solution, as shown in fig. 4, is substantially the same as in example 1, except that: the number of the emitting light sources is 2, the light source 1 is used for emitting laser with the wavelength of 620-970 nm, and the light source 2 is used for emitting laser with the wavelength of 300-600 nm.
Example 3
A method for measuring particle parameters in a solution comprises the following steps:
(1) HeLa cells (adherent cells) and HL60 cells (suspension cells) were counted using a Countstar BioTech automatic cell counter IC1000 (Countstar, shanghai, china) and serially diluted in fresh medium and saline solution (SPSS), respectively, prior to OD measurement to give four diluted samples (test solutions), each comprising 5 or more concentrations:
first set of test solutions: HL60 cells, taking RPMI-1640 cell culture solution as a medium;
second set of test solutions: hela cells, which use DMEM cell culture medium as medium;
third group of test solutions: HL60 cells, physiological saline as medium;
fourth set of test solutions: hela cells, which use physiological saline as a medium;
(2) The first to fourth sets of solutions to be tested were each independently inputted into the test apparatus in example 1, and the irradiation wavelength of the light source was set to be 750nm.
Example 4
A method for measuring particle parameters in solution, which is substantially the same as in example 1, except that: the irradiation wavelength in example 1 was replaced with 850nm.
Example 5
A method for measuring particle parameters in solution, which is substantially the same as in example 1, except that: the irradiation wavelength in example 1 was replaced with 970nm.
Comparative example 1
A method for measuring particle parameters in solution, which is substantially the same as in example 1, except that: the irradiation wavelength in example 1 was replaced with 410nm.
Comparative example 2
A method for measuring particle parameters in solution, which is substantially the same as in example 1, except that: the irradiation wavelength in example 1 was replaced with 560nm.
Comparative example 3
A method for measuring particle parameters in solution, which is substantially the same as in example 1, except that: the irradiation wavelength in example 1 was replaced with 580nm.
The absorbance of each of the solutions to be measured in examples 3 to 5 and comparative examples 1 to 3 was recorded, and a fitted curve of the concentration and absorbance was drawn as shown in fig. 1.
As can be seen from fig. 1, the concentrations of HeLa cells and HL60 cells at each wavelength plotted against the corresponding optical densities all exhibited a significant linear positive correlation. The correlation coefficient (R 2 ) All are noted in Table 1, the optical densities measured using different media and cells all show good linear relationship with cell concentration, and the linear correlation coefficients obtained at 6 wavelengths for the four cell-media combinations are all substantially largeAt 0.983, the correlation coefficient of the linear regression curve fitted with physiological saline as the control group was slightly higher than that of the culture solution.
There was some deviation between the standard curves obtained using different cell types and vehicle tests. Wherein, the standard curves under different conditions have the largest deviation when the wavelengths are 410, 560 and 580nm (sensitive wavelengths). The standard curves are closest to each other at wavelengths of 750, 850, 970nm (stable wavelengths), but the gap is not completely eliminated. This demonstrates that the selection of wavelengths in the range of 620-970 nm to measure the optical density of cells results in a standard curve that is better compatible with different cells and media.
TABLE 1
Example 6 bubble detection
The method for measuring particle parameters in solution adopts the device in the embodiment 1 to detect whether bubbles exist in cell suspension, and the testing process is as follows:
(1) The irradiation wavelength of the light source of the device in example 1 was set to 850nm, and the output voltage value (positive voltage) was recorded as v1=3.2v, using air as the detection target.
(2) Distilled water was placed in the channel of example 1 under the same conditions as in step (1) as the detection object, and the output voltage value was recorded as v2=2.8v.
(3) Placing the cell suspension in the channel of example 1 under the same conditions as in step (1), and outputting a voltage value of v=0.8v; since V satisfies V < (v1+v2)/2=3v, the bubble is not contained.
(4) Air is pumped in by adopting a peristaltic pump, and after bubbles are generated in the channel in the embodiment 1, the step (3) is repeated, wherein the output voltage value is V=3.18V, and V is more than or equal to (V1+V2)/2=3V.
Example 7 detection of the presence of phenol Red indicator
A method for measuring particle parameters in a solution comprises culturing physiological saline cells and containing Hela cells as a solution to be measured (wherein phenol red is not contained,and the cell concentration was 6.18X10 5 Per mL) and was input into the test apparatus in example 2, the irradiation wavelength of the light source 1 was set to 850nm, and the irradiation wavelength of the light source 2 was set to 570nm.
Substituting absorbance (0.865) obtained by detection of the light source 1 into a standard curve which is formed by the cells and is obtained by fitting after testing OD values of standard solution without concentration, and calculating the concentration of HeLa cells in the solution to be tested to be 6.08X10 5 individual/mL;
substituting the calculated concentration into a cell concentration equivalent absorbance curve of HELA cells at 570nm wavelength to obtain equivalent absorbance of 3.3;
let y=absorbance detected by light source 2/equivalent absorbance=0.12;
because y is less than or equal to 0.2, the solution to be measured does not contain a phenol red indicator.
Example 8 detection of the presence of phenol Red indicator
A method for measuring particle parameters in solution comprises culturing DMEM cell in which Hela cells are contained as the solution to be measured (containing phenol red with cell concentration of 8.2X10) 5 Per mL) and was input into the test apparatus in example 2, the irradiation wavelength of the light source 1 was set to 850nm, and the irradiation wavelength of the light source 2 was set to 570nm.
Substituting the absorbance (1.1) obtained by the detection of the light source 1 into y in a standard curve y=1.196 e-7x+0.1371, calculating x, and obtaining the concentration of Hela cells in the solution to be detected as 8.2 multiplied by 10 5 individual/mL;
substituting the calculated concentration into a cell concentration equivalent absorbance curve (shown in figure 5) of Hela cells at 570nm wavelength to obtain equivalent absorbance of 4.25;
let y=absorbance detected by light source 2/equivalent absorbance=2.04/4.25=0.48;
because y is more than 0.2, the solution to be measured contains a phenol red indicator, and y is substituted into a calculation formula corresponding to a fitting curve shown in fig. 6 to obtain the pH value (namely the value of x in the formula) of the solution to be measured: 7.05.
in example 8, the fit curve was obtained by the following method when determining the pH: the absorbance of the phenol red solution with the standard pH value is measured by using a spectrometer, and the absorbance is obtained by piecewise fitting through a linear regression method, the excitation wavelength of the spectrometer is 200 nm-1000 nm, and a fitting curve corresponding to any value in the wavelength range can be obtained, wherein the fitting curve of 570nm wavelength is shown as a graph in figure 6, the fitting curve corresponding to 3 < pH < 7 is shown as a graph in figure 6, and the fitting curve corresponding to 7 < pH < 10 is shown as a graph in figure 6 b.
Example 9
A method for measuring particle parameters in a solution comprises the following steps:
(1) K562 cells (suspension cells) and a549 cells (adherent cells) were counted using a Countstar BioTech automatic cell counter IC1000 (Countstar, shanghai, china) and serially diluted in fresh medium and saline solution (SPSS) respectively before OD measurement to obtain four groups of diluted samples (test solutions) each comprising 7 concentrations:
first set of test solutions: the K562 cells use physiological saline as a medium;
second set of test solutions: the K562 cells use RPMI-1640 cell culture solution as a medium;
third group of test solutions: a549 cells use physiological saline as a medium;
fourth set of test solutions: a549 cells use DMEM cell culture medium as medium;
(2) The first to fourth groups of solutions to be tested were each independently inputted into the test device in example 1, and the irradiation wavelength of the light source was set to 850nm, the sensor voltage value was recorded and the absorbance value of each solution to be tested was tested, and a fitted curve at 850nm wavelength was drawn, as shown in fig. 2, the absorbance values of cells in different solutions linearly increased with the increase of the cell concentration, and the fitted curve under each solution condition had a high regression coefficient R 2 Value of R 2 Are all greater than 0.99. This indicates that the measurements performed are highly reproducible and accurate.
In practical application, as long as the absorbance value of the solution to be measured and the cell concentration can establish exact correlation, a fitting curve obtained by experiments can be utilized and used for measuring the subsequent concentration.
Finally, it should be noted that: the above embodiments are only for illustrating the technical solution of the present invention, and are not limiting; although the invention has been described in detail with reference to the foregoing embodiments, it will be understood by those of ordinary skill in the art that: the technical scheme described in the foregoing embodiments can be modified or some technical features thereof can be replaced by equivalents; such modifications and substitutions do not depart from the spirit and scope of the technical solutions of the embodiments of the present invention.
Claims (10)
1. A method for measuring a particle parameter in a solution, comprising: and using laser with the wavelength of 620-970 nm as a first light source to irradiate the solution to be measured, using an optical receiver to receive the light passing through the solution to be measured, and judging whether the solution to be measured contains bubbles and/or the concentration of particles in the solution to be measured.
2. The method for measuring particle parameters in solution according to claim 1, wherein the optical receiver is a photosensor and outputs a voltage; the judging comprises the following steps:
when the voltage is positive voltage, if the detected voltage value is larger than (V1 +V2)/2, the air bubble is contained, otherwise, the air bubble is not contained;
when the voltage is a negative voltage, if the detected voltage value is smaller than (V1 +V2)/2, the air bubble is contained, otherwise, the air bubble is not contained;
wherein V1 is a voltage value when air is detected under the same condition, and V2 is a voltage value when water is detected under the same condition.
3. The method for measuring particle parameters in a solution according to claim 1 or 2, wherein the solution to be measured is a culture solution containing animal cells, and the particles are animal cells in the culture solution, preferably, the animal cells are suspended cells.
4. A method for measuring particle parameters in solution according to claim 3, wherein the process of measuring animal cells in the culture solution comprises:
(1) Testing the culture solution with known animal cell concentration under the wavelength to obtain corresponding OD value, and drawing a fitting curve of the relevant concentration and the OD value;
(2) Substituting the OD value measured by the solution to be measured under the wavelength into the fitting curve to obtain the concentration of the animal cells in the solution to be measured.
5. The method for measuring a particle parameter in a solution according to claim 3 or 4, further comprising: irradiating the solution to be detected by using laser with the wavelength of 300-600 nm as a second light source, and judging whether the culture solution contains a phenol red indicator or not;
preferably, the second light source is a laser with a wavelength of 320-470 nm or 500-580 nm;
more preferably, a laser having a wavelength of 560.+ -.20 nm is used as the second light source.
6. The method for measuring particle parameters in solution according to claim 5, wherein the measuring process comprises:
the absorbance obtained by irradiating the solution to be measured by using laser with the wavelength of 300-600 nm as a second light source is marked as A;
calculating the equivalent absorbance of the cell concentration corresponding to the solution to be detected under the wavelength used by the second light source, and marking as B;
if A/B is more than 0.2, the solution to be measured contains a phenol red indicator, otherwise, the solution to be measured does not contain the phenol red indicator.
7. The method for measuring particle parameters in a solution according to any one of claims 3 to 6, wherein if the culture medium contains a phenol red indicator, the method further comprises: irradiating the solution to be detected by using laser with the wavelength of 300-600 nm as a second light source, and judging the pH value of the culture solution;
preferably, the second light source is a laser with a wavelength of 320-470 nm or 500-580 nm;
more preferably, a laser having a wavelength of 560.+ -.5 nm is used as the second light source.
8. The method for measuring particle parameters in solution according to claim 7, wherein the measuring process comprises:
the absorbance obtained by irradiating the solution to be measured by using laser with the wavelength of 300-600 nm as a second light source is marked as A;
calculating the equivalent absorbance of the cell concentration corresponding to the solution to be detected under the wavelength used by the second light source, and marking as B;
let a/b=c, substituting C into the absorbance standard curve of the standard pH phenol red solution to obtain the pH of the culture solution.
9. The method according to any one of claims 3 to 8, wherein the number of the lasers is 2 or more, for simultaneously measuring the concentration of animal cells and other parameters in the culture solution;
the other parameters include: whether the culture solution contains bubbles, whether the culture solution contains a phenol red indicator and/or the pH value of the culture solution.
10. A device for measuring a particle parameter in a solution, comprising: a channel for circulating the solution to be measured, a light source for emitting laser light and irradiating the solution to be measured, and an optical receiver for receiving light passing through the solution to be measured; the optical path formed by the light source is perpendicular to the channel;
preferably, the number of the light sources is more than 2;
more preferably, the equivalent diameter of the cross section of the channel is 0.5 mm-15 mm, and the equivalent diameter variation value of the cross section in the flow direction is less than 1mm, and the transmittance of the laser passing through the channel is more than 80%.
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