A cell characteristic detection device and detection method thereof

Abstract

The present invention relates to a biological detection device, and in particular to a cell characteristic detection device and a detection method thereof. Detection device: a light source emitter is connected to an incident light transmission medium; a light reflection layer is arranged near the light outlet of the incident light transmission medium and the light entrance of the reflected light transmission medium; the reflected light transmission medium is connected to a light intensity acquisition and detection element. Detection method: a light source is applied to obtain a first light intensity of a sample to be tested before inoculating cells; a light source is applied to obtain a second light intensity emitted by the sample to be tested after inoculating cells; cell characteristics are determined by comparing the first light intensity and the second light intensity. The present invention utilizes changes in light reflection intensity to accurately and qualitatively and quantitatively determine cell concentration, state and type, and can more accurately reflect the true biological characteristics of the sample compared to traditional methods.

Description

Technical Field

[0001] The present invention relates to a biological detection device, and in particular to a cell characteristic detection device and a detection method thereof. Background Art

[0002] In biotechnology and medical research, it is crucial to quickly and accurately determine the concentration, state and type of cells. In the prior art, the determination of cell characteristics usually relies on microscopic observation or specific biochemical markers, which are not only complicated to operate, but also time-consuming.

[0003] In fermentation engineering, monitoring the cell concentration in the culture tank is of great significance for understanding the state of the fermentation process and regulating the fermentation conditions. The light emitting and receiving devices required by the existing optical density detector are difficult to install, and the optical path is insufficient, resulting in low measurement accuracy, slow response speed, and complex operation, which is difficult to meet the needs of practical applications. Summary of the invention

[0004] 1. Technical issues to be resolved

[0005] In view of the above-mentioned shortcomings and deficiencies of the prior art, the present invention provides a cell characteristic detection device that can quickly and accurately determine the concentration, state and type of cells and simplify the operation process.

[0006] Correspondingly, the present invention also provides a method for detecting cell characteristics.

[0007] (II) Technical solution

[0008] In order to achieve the above object, the main technical solutions adopted by the present invention include:

[0009] In a first aspect, the present invention provides a cell characteristic detection device, which comprises: a light source emitter, an incident light transmission medium, a reflected light transmission medium, a light reflection layer and a light intensity acquisition and detection element,

[0010] The light source transmitter is connected to the incident light transmission medium;

[0011] The light reflecting layer is arranged near the light outlet of the incident light transmission medium and the light inlet of the reflected light transmission medium;

[0012] The reflected light transmission medium is connected to the light intensity acquisition and detection element.

[0013] Optionally, it also includes: a data processing module and / or a display module,

[0014] The data processing module is signal-connected to the light intensity acquisition and detection element;

[0015] The display module is connected to the data processing module via signals.

[0016] The present invention transmits the light emitted by the light source transmitter to the light reflection layer through the incident light transmission medium, reflects the light through the light reflection layer, and transmits the reflected light back to the light intensity acquisition and detection element through the reflective light transmission medium, shortens the optical detection distance by reflecting the light, and realizes the detection of the effective optical path. More specifically, during the measurement, the light reflection layer is immersed in the sample liquid.

[0017] The light source emitter of the present invention can be, but is not limited to, an LED or a laser to provide stable and uniform light.

[0018] The light intensity acquisition and detection element may be, but is not limited to, a photodetector such as a photodiode or a photomultiplier tube to detect the intensity of reflected light with high sensitivity.

[0019] The display module and the data processing module may be, but are not limited to, an integrated microprocessor or computer system to process signals and display measurement results through a graphical user interface.

[0020] The data processing module processes the light intensity through the formulas of Formula 1 and Formula 2, and the light intensity obtained by the detection element, wherein I0 represents the first light intensity; I n Represents the second light intensity at a certain moment;

[0021] Optionally, the incident light transmission medium and the reflected light transmission medium are one or a combination of more than one of the following: optical fiber, crystal, and quartz.

[0022] When the incident light transmission medium and the reflected light transmission medium are optical fibers, they are presented in the form of an optical fiber bundle structure, and the light reflection layer is at a certain distance from the light exit of the optical fiber bundle.

[0023] Optionally, it further comprises a protective cover, wherein the incident light transmission medium and the reflected light transmission medium are arranged in the protective cover, and the lower end surface of the protective cover is arranged to be light-transmissive.

[0024] The light reflecting layer is arranged below the protective cover, and there is a space between the protective cover and the light reflecting layer to allow the sample liquid to flow through.

[0025] The light passes through the protective cover and then through the sample liquid to reach the light reflecting layer, and the reflected light is transmitted back to the reflecting light transmission medium.

[0026] Optionally, the cell characteristics include one or a combination of two or more of cell concentration, cell state and cell type;

[0027] When the sample liquid contains a single cell or more than one cell, the light reflection intensity of the sample liquid will change. At the same time, the light reflection intensity of organisms or cells of different growth states and types will also change. Therefore, the concentration, state and type of cells and other organisms can be qualitatively and quantitatively determined by the light reflection intensity.

[0028] The cell characteristics include cell characteristics of a single cell or an organism composed of more than one cell.

[0029] In a second aspect, the present invention further provides a method for detecting cell characteristics, which comprises the following steps:

[0030] S1: applying a light source to obtain a first light intensity of the sample to be tested before inoculating cells;

[0031] S2: applying a light source to obtain a second light intensity emitted by the sample to be tested after inoculation of cells;

[0032] S3: Determine the cell characteristics by comparing the first light intensity and the second light intensity.

[0033] Optionally, the method for acquiring the first light intensity and the second light intensity includes: determining by intensity of reflected light passing through the sample liquid.

[0034] Optionally, the cell characteristics are further characterized by Formula 1:

[0035] A=K[-log(I n / I0)] Formula 1;

[0036] Among them, A represents the cell coefficient, which is used to characterize cell characteristics;

[0037] I0 represents the first light intensity;

[0038] I n represents the second light intensity at a certain moment;

[0039] K is a constant;

[0040] Among them, K can be determined by performing multiple tests on samples with known cell characteristics to determine the K value.

[0041] Optionally, the cell characteristics are also qualitatively and / or quantitatively determined by formula 2:

[0042] C n =A n +(1-α)A (n-1) Formula 2:

[0043] Among them, Cn represents the cell characteristic value;

[0044] α is the filter coefficient (in the range of 0 to 1);

[0045] A n It indicates that the second light intensity is obtained by the second measurement, and the cell coefficient A is obtained according to formula 1;

[0046] A (n-1) It indicates that the second light intensity is obtained by the first measurement and the cell coefficient A is obtained according to formula 1.

[0047] The α value can be determined by performing multiple tests on samples with known cell characteristics.

[0048] In organisms including cells or bacteria, during the growth process including fermentation, bubbles formed due to various reasons (such as gas supply) and large particles caused by the aggregation of organisms including cells or bacteria will interfere with the concentration detection of the organism, causing fluctuations and deviations in the detection of cell characteristics such as the concentration of the organism. In order to remove the interference, the present invention removes the interference by low-pass filtering to ensure the accuracy of the measured value.

[0049] (III) Beneficial effects

[0050] The beneficial effects of the present invention are:

[0051] 1. Improve accuracy and reliability: The present invention utilizes the change in light reflection intensity to accurately and qualitatively and quantitatively determine the concentration, state and type of cells, which can more accurately reflect the true biological characteristics of the sample compared to traditional methods.

[0052] 2. Rapid response: The present invention utilizes the intensity of light reflection to make the detection process rapid, thereby significantly shortening the time from sample preparation to result acquisition.

[0053] 3. Strong adaptability: The detection device of the present invention can adapt to different experimental environments and requirements, such as using different incident light transmission media (optical fiber, crystal, quartz) to adapt to different sample characteristics.

[0054] 4. Enhanced stability: Through low-pass filtering technology, interference caused by bubbles, aggregation and other factors is effectively eliminated, ensuring the stability and reliability of the data. BRIEF DESCRIPTION OF THE DRAWINGS

[0055] Figure 1 It is a schematic diagram of the structure of the cell characteristic detection device of Example 1 and Example 2 of the present invention;

[0056] 1. Incident light transmission medium; 2. Reflected light transmission medium; 3. Protective cover; 4. Reflective layer; 5. Light intensity acquisition and detection element. DETAILED DESCRIPTION

[0057] In order to better explain the present invention, so as to facilitate understanding, exemplary embodiments of the present invention will be described in more detail below. Although exemplary embodiments of the present invention are shown below, it should be understood that the present invention can be implemented in various forms and should not be limited by the embodiments set forth herein. On the contrary, these embodiments are provided in order to be able to more clearly and thoroughly understand the present invention, and to be able to fully convey the scope of the present invention to those skilled in the art.

[0058] Reference to "embodiment" herein means that the specific features, structures or characteristics described in conjunction with the embodiment may be included in at least one embodiment of the present application. The term "embodiment" appearing in various places in the specification does not necessarily refer to the same embodiment, nor does it particularly limit its independence or association with other embodiments. In principle, in the present application, as long as there is no technical contradiction or conflict, the various technical features mentioned in the embodiments can be combined in any way to form a corresponding implementable technical solution.

[0059] Unless otherwise defined, the technical terms used in this document have the same meanings as those generally understood by those skilled in the art to which this application belongs; the use of relevant terms in this document is only for describing specific embodiments and is not intended to limit this application.

[0060] In the description of this application, the term "and / or" is an expression used to describe the logical relationship between objects, indicating that three relationships may exist, for example, A and / or B, which means: A exists, B exists, and A and B exist at the same time. In addition, the character " / " in this article generally indicates that the objects before and after are in an "or" logical relationship.

[0061] In the present application, terms such as “first” and “second” are merely used to distinguish one entity or operation from another entity or operation, and do not necessarily require or imply any actual relationship of quantity, priority or sequence between these entities or operations.

[0062] Without further limitations, in this application, the words "include", "comprises", "has" or other similar expressions used in the sentences are intended to cover non-exclusive inclusion. These expressions do not exclude the presence of additional elements in the process, method or product including the elements, so that the process, method or product including a series of elements may include not only those limited elements, but also other elements not explicitly listed, or also include elements inherent to such process, method or product.

[0063] Similar to the understanding in the Examination Guidelines, in this application, expressions such as "greater than", "less than", "exceed" and the like are understood to exclude the number itself; expressions such as "above", "below", "within" and the like are understood to include the number itself. In addition, in the description of the embodiments of this application, "multiple" means more than two (including two), and similar expressions related to "multiple" are also understood in this way, such as "multiple groups", "multiple times", etc., unless otherwise clearly and specifically limited.

[0064] In the description of the embodiments of the present application, space-related expressions used, such as "center", "longitudinal", "lateral", "length", "width", "thickness", "up", "down", "front", "back", "left", "right", "vertical", "horizontal", "vertical", "top", "bottom", "inside", "outside", "clockwise", "counterclockwise", "axial", "radial", "circumferential", etc., indicate the orientation or position relationship based on the orientation or position relationship shown in the specific embodiments or drawings, and are only for the convenience of describing the specific embodiments of the present application or facilitating the reader's understanding, and do not indicate or imply that the referred device or component must have a specific position, a specific orientation, or be constructed or operated in a specific orientation, and therefore cannot be understood as a limitation on the embodiments of the present application.

[0065] Unless otherwise expressly specified or limited, in the description of the embodiments of the present application, the terms such as "install", "connect", "connect", "fix", "set", etc. used should be understood in a broad sense. For example, the "connection" can be a fixed connection, a detachable connection, or an integrated setting; it can be a mechanical connection, an electrical connection, or a communication connection; it can be a direct connection, or it can be indirectly connected through an intermediate medium; it can be the internal connection of two elements or the interaction relationship between two elements. For technicians in the technical field to which the present application belongs, the specific meanings of the above terms in the embodiments of the present application can be understood according to the specific circumstances.

[0066] Example 1

[0067] like Figure 1 As shown, this embodiment provides a cell feature detection device, which comprises: a light source transmitter, an incident light transmission medium 1, a reflected light transmission medium 2, a light reflection layer 4 and a light intensity acquisition and detection element 5,

[0068] The light source transmitter is connected to the incident light transmission medium 1;

[0069] The light reflecting layer 4 is arranged near the light outlet of the incident light transmission medium 1 and the light entrance of the reflected light transmission medium 2; the reflected light transmission medium 2 is connected to the light intensity acquisition and detection element 5.

[0070] In this embodiment, the light emitted by the light source transmitter is transmitted to the light reflection layer 4 through the incident light transmission medium 1, and the light is reflected by the light reflection layer 4. The reflected light is transmitted back to the light intensity acquisition and detection element 5 through the reflective light transmission medium 2. By reflecting the light, the optical detection distance is shortened and the detection of the effective optical path is realized. More specifically, during the measurement, the light reflection layer 4 is immersed in the sample liquid.

[0071] Example 2

[0072] like Figure 1 As shown, this embodiment provides a more specific cell feature detection device relative to the embodiment 1, which includes: a light source transmitter, an optical fiber for transmitting incident light, an optical fiber for transmitting reflected light, a light reflection layer 4, a light intensity acquisition and detection element 5, a data processing module and a display module.

[0073] The optical fiber for transmitting the incident light and the optical fiber for transmitting the reflected light are presented in the form of an optical fiber bundle and are arranged in the protective cover 3; the lower end surface of the protective cover 3 is arranged to be light-transmissive;

[0074] The light emitted by the light source transmitter is propagated through the optical fiber used to transmit the incident light;

[0075] The optical fiber for transmitting the incident light is connected to the light intensity acquisition and detection element 5 .

[0076] The light reflecting layer 4 is at a certain distance from the light outlet of the optical fiber bundle.

[0077] The light reflecting layer 4 is below the protective cover 3, and there is a space between the protective cover 3 and the light reflecting layer 4 to allow the sample liquid to flow through. The light reflecting layer 4 can be connected to the protective cover 3 through a connecting rod, or can be independently arranged in the container where the sample to be tested is placed.

[0078] The data processing module is connected to the light intensity acquisition and detection element 5 by signal;

[0079] The display module is connected to the data processing module via signals.

[0080] The present invention transmits the light emitted by the light source transmitter to the light reflection layer 4 through the optical fiber used to transmit the incident light, and the light is reflected by the light reflection layer 4. The reflected light is transmitted back to the light intensity acquisition and detection element 5 through the optical fiber used to transmit the reflected light. By reflecting the light, the optical detection distance is shortened and the detection of the effective optical path is realized. More specifically, the light reflection layer 4 is immersed in the sample liquid during measurement.

[0081] The data processing module processes the light intensity by the formulas of Formula 1 and Formula 2, and obtains the light intensity obtained by the detection element 5, wherein I0 represents the first light intensity; I n Represents the second light intensity at a certain moment;

[0082] In this embodiment, the light emitted by the protective cover 3 is irradiated into the culture liquid to detect the cultured cells in the culture tank. The light source emits light to irradiate the liquid in the culture tank, and reaches the reflective surface after passing through the set optical path. The light reflected back by the reflective surface then passes through the optical fiber to reach the photoelectric sensor light intensity acquisition and detection element 5 signal, and the photoelectric sensor light intensity acquisition and detection element 5 signal is converted into an electrical signal and transmitted to the data processing module. The data processing module processes the received electrical signal, calculates the characteristic values ​​such as the concentration of the liquid, and displays it through the display unit.

[0083] Example 3

[0084] This embodiment provides a method for detecting the concentration of an Escherichia coli sample solution, the steps of which are:

[0085] S1 Preliminary experiment: prepare multiple cultures of E. coli with different concentrations as preliminary experimental samples;

[0086] The culture broth control sample without inoculation of E. coli was sterile culture medium;

[0087] S2 Preparation of the cell feature detection device in Example 1 or Example 2: Using an LED light source with a wavelength of 600 nm as a light source emitter;

[0088] S3 Optical intensity measurement: The optical intensity reflected by the pre-experimental sample and the control sample is measured by the cell characteristic detection device.

[0089] S4 Substitute the optical intensity measured by the pre-experimental sample into I in formula 1 n Substitute the optical intensity obtained from the pre-experimental sample into I0 in formula 1, and the obtained A is recorded as A n-1 ,

[0090] After the pre-experimental sample is cultured for a period of time, the optical intensity measured by the cell characteristic detection device is substituted into I in formula 1. n Substitute the optical intensity obtained from the pre-experimental sample into I0 in formula 1, and the obtained A is recorded as A n , the obtained A n-1 and A n Substitute into formula 2 to get C n ,

[0091] A=K[-logI n / I0] Formula 1;

[0092] C n =A n +(1-α)A (n -1) Formula 2;

[0093] Among them, Cn represents the cell characteristic value;

[0094] α represents the filter coefficient in the range of 0 to 1;

[0095] A n It represents the cell coefficient A obtained by formula 1 in the second test;

[0096] A (n-1) Represents the cell coefficient A obtained by formula 1 in the first detection.

[0097] Each pre-experimental sample corresponding to each concentration can obtain an optical intensity; thus, multiple binary linear equations can be obtained, and the K value and A value can be calculated and corrected, and a relationship curve between concentration and A value can be drawn; after calculation, the cell coefficient K of Escherichia coli is set to 0.85 and α is 0.2;

[0098] S5 measured the optical intensity of the sample to be tested by the cell characteristic detection device to be 1800 units, which was substituted into I in formula 1. n , the optical intensity of the pre-experimental sample is 2000 units, and it is substituted into I0 in formula 1 to obtain A n-1 =0.038893; Similarly, the optical intensity 1500 units obtained from the second measurement is substituted into I n , A obtained in formula 1 n =0.10625, which can be used to further analyze the concentration of E. coli. Under given experimental conditions, the presence of bacteria causes a decrease in light intensity, which reflects the concentration level of bacteria. Higher or lower A values ​​indicate different cell concentrations, which is useful for adjusting culture conditions or evaluating bacterial growth status.

[0099] Substitute into calculation formula 2, C n =0.1373644 represents the cell concentration after considering the fluctuations of continuous measurements. This value more accurately reflects the actual characteristics of the cells and reduces the impact of accidental errors caused by changes within the measurement interval. In this way, using formula 2 can effectively smooth the data and provide more stable and reliable measurement results, especially in long-term monitoring or biological processes with large dynamic changes.

[0100] Example 4

[0101] This embodiment provides a method for detecting differentiated tumor cells and undifferentiated tumor cells. The steps are the same as those in Embodiment 3. The difference lies in the difference in K value and α value. n Indicated are tumor cells in different differentiation states.

[0102] In this embodiment, tumor cells with different degrees of differentiation and A values, as well as C nThe relationship curve of the value can be obtained by measuring the A value or C value of the sample to be tested. n The values ​​can qualitatively and quantitatively determine the degree of differentiation of tumor cells.

[0103] Finally, it should be noted that although the above embodiments have been described in the specification and drawings of this application, this does not limit the scope of patent protection of this application. All technical solutions generated by replacing or modifying equivalent structures or equivalent processes based on the essential concept of this application using the contents recorded in the specification and drawings of this application, as well as directly or indirectly implementing the technical solutions of the above embodiments in other related technical fields, are included in the scope of patent protection of this application.

Claims

1. A cell characteristic detection device, characterized in that: It includes: A light source transmitter, an incident light transmission medium (1), a reflected light transmission medium (2), a light reflection layer (4) and a light intensity acquisition and detection element (5), The light source transmitter is connected to the incident light transmission medium (1); The light reflecting layer (4) is arranged near the light outlet of the incident light transmission medium (1) and the light inlet of the reflected light transmission medium (2); The reflected light transmission medium (2) is connected to a light intensity acquisition and detection element (5).

2. The cell characteristic detection device according to claim 1, characterized in that: It also includes: a data processing module and / or a display module, The data processing module is signal-connected to the light intensity acquisition and detection element (5); The display module is connected to the data processing module via signals.

3. The cell characteristic detection device according to claim 1, characterized in that: The incident light transmission medium (1) and the reflected light transmission medium (2) are one or a combination of more than one of the following: optical fiber, crystal, and quartz.

4. The cell characteristic detection device according to claim 1, characterized in that: It also includes a protective cover (3), the incident light transmission medium (1) and the reflected light transmission medium (2) are arranged in the protective cover (3), and the lower end surface of the protective cover (3) is arranged to be light-transmissive. The light reflecting layer (4) is arranged below the protective cover (3), and there is a space between the protective cover (3) and the light reflecting layer (4) to allow the sample liquid to flow through.

5. The cell characteristic detection device according to any one of claims 1 to 4, characterized in that: The cell characteristics include one or a combination of two or more of cell concentration, cell state and cell type; The cell characteristics include cell characteristics of a single cell or an organism composed of more than one cell.

6. A method for detecting cell characteristics, characterized in that: It includes the following steps: S1: applying a light source to obtain a first light intensity of the sample to be tested before inoculating cells; S2: applying a light source to obtain a second light intensity emitted by the sample to be tested after inoculation of cells; S3: Determine the cell characteristics by comparing the first light intensity and the second light intensity.

7. The method for detecting cell characteristics according to claim 6, characterized in that: The first light intensity and the second light intensity are intensities of reflected light after the incident light passes through the sample to be measured and is reflected.

8. The method for detecting cell characteristics according to claim 6, characterized in that: It also characterizes the cell characteristics by formula 1: A=K[-log(I n / I0)] Formula 1; Among them, A represents the cell coefficient, which is used to characterize cell characteristics; I0 represents the first light intensity; I n represents the second light intensity at a certain moment; K is a constant.

9. The method for detecting cell characteristics according to claim 8, characterized in that: It also qualitatively and / or quantitatively characterizes the cell characteristics by formula 2: C n = A n + (1 - α)A (n-1) Formula 2; Among them, C n Represents the cell characteristic value; α is represented as the filter coefficient; A n It indicates that the second light intensity is obtained by the second measurement, and the cell coefficient A is obtained according to formula 1; A (n-1) It indicates that the second light intensity is obtained by the first measurement and the cell coefficient A is obtained according to formula 1.

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