Bioparticle screening assay

Abstract

A method for screening and detecting biological particles includes the following steps: providing a biological particle screening and detection system, comprising a flow channel, an optical detection component, and a carrier disk, wherein the optical detection component provides a detection optical path that penetrates the flow channel, and the optical detection component is used to sense a target biological particle; providing a sample; diluting the sample to form a test solution; controlling the test solution to flow into the flow channel from an inlet end; and when the optical detection component senses the target biological particle in the test solution, the carrier disk is controlled to move to an outlet end of the flow channel, wherein the carrier disk contains a diluent, and a portion of the solution containing the target biological particle in the test solution is controlled to be discharged from the outlet end and then loaded onto the carrier disk to dilute the portion of the solution.

Description

Technical Field

[0001] This invention relates to biochemical detection systems; in particular, it relates to a method for screening and detecting biological particles. Background Technology

[0002] Traditional biochemical detection methods involve manually analyzing biological samples using techniques such as fluorescence staining and immunoprecipitation. However, these methods are limited by the equipment and skills required for manual operation and necessitate large sample volumes (e.g., blood, other bodily fluids, hair, or nails), making it difficult to perform micro-scale detection.

[0003] In addition to the difficulty of performing trace detection manually, when the number of biological particles in a biochemical sample is large, it will increase the difficulty and reduce the accuracy of identifying or screening target biological particles. Therefore, how to provide a biological particle screening and detection method that can perform trace detection and improve the accuracy of identifying or screening target biological particles is an urgent problem to be solved. Summary of the Invention

[0004] In view of this, the purpose of the present invention is to provide a biological particle screening and detection method that can perform trace detection and improve the accuracy of identifying or screening target biological particles.

[0005] To achieve the above objectives, the present invention provides a biological particle screening and detection method comprising the following steps:

[0006] A biological particle screening and detection system is provided, comprising a flow channel, an optical detection component and a carrier disk, wherein the optical detection component provides a detection optical path that penetrates the flow channel, and the optical detection component is used to sense a target biological particle;

[0007] Provide a sample;

[0008] The sample was diluted to form a test solution;

[0009] The test solution is controlled to flow into the channel from one inlet end; and

[0010] When the optical detection component senses the target biological particles in the test solution, the carrier disk is controlled to move to an outlet end of the flow channel, wherein a diluent is contained in the carrier disk, and a portion of the test solution containing the target biological particles is discharged from the outlet end and then loaded onto the carrier disk to dilute the portion of the solution.

[0011] The advantage of this invention is that the biological particle screening and detection method of this invention can perform trace detection, and by diluting the sample to form the test solution, when the number of biological particles in the sample is large, it can avoid the influence of clustered biological particles or excessively high concentrations of the sample on the accuracy of the optical detection component in sensing the target biological particles in the test solution, thereby improving the accuracy and stability of trace detection. Attached Figure Description

[0012] Figure 1 This is a flowchart of a preferred embodiment of the biological particle screening and detection method of the present invention.

[0013] Figure 2 This is a block diagram of a biological particle screening and detection system according to a preferred embodiment of the present invention.

[0014] Explanation of reference numerals in the attached figures:

[0015] 1: Biological particle screening and detection system

[0016] 10: Flow channel

[0017] 12: Entry point

[0018] 14: Export end

[0019] 20: Optical inspection components

[0020] 22: Excitation Light Source

[0021] 24: Light sensor

[0022] 30: Carrier disk

[0023] 40: Sample source

[0024] 50: Sample supply device

[0025] 60: Carrier unit

[0026] R: Detection optical path

[0027] S02, S04, S06, S08, S10: Steps Detailed Implementation

[0028] To more clearly illustrate the present invention, preferred embodiments are described in detail below with reference to the accompanying drawings. Please refer to... Figure 1 The diagram shown is a flowchart of a preferred embodiment of a biological particle screening and detection method according to the present invention. The biological particle screening and detection method includes the following steps:

[0029] Step S02: A biological particle screening and detection system 1 is provided, comprising a flow channel 10, an optical detection component 20, and a carrier disk 30. The optical detection component 20 provides a detection optical path R that penetrates the flow channel 10. The optical detection component 20 is used to sense a target biological particle. In this embodiment, the biological particle screening and detection method is achieved through, for example... Figure 2 The biological particle screening and detection system 1 shown is used for execution. In other embodiments, the biological particle screening and detection method can also be executed by other biological particle screening and detection systems, and is not limited thereto.

[0030] Step S04: Provide a sample; the sample may be a blood sample, urine sample, pleural effusion, ascites, spinal fluid, cerebrospinal fluid, or amniotic fluid.

[0031] Step S06 involves diluting the sample to form a test solution. In this embodiment, the test solution is further mixed to ensure uniform distribution of biological particles. Further, step S06 includes diluting the sample by 5X to 100X. In one embodiment, when the blood sample is 8 ml, 32 ml of diluent is added to dilute the sample by 5X. In another embodiment, when the blood sample is 4 ml, 36 ml of diluent is added to dilute the sample by 10X. In another embodiment, when the blood sample is 1 ml, 39 ml of diluent is added to dilute the sample by 40X. In yet another embodiment, when the blood sample is 1 ml, 99 ml of diluent is added to dilute the sample by 100X. By diluting the blood sample by 5X to 100X, not only can excessively high blood sample concentrations be avoided, which would make subsequent biological particle acquisition more difficult, but excessively low blood sample concentrations can also be avoided, which would result in a significant time commitment for subsequent biological particle screening and detection.

[0032] It should be further noted that the aforementioned diluents are buffer solutions that can maintain the pH of the sample, or cell culture media suitable for culturing cells, such as: Tris buffer, Phosphate buffer, HEPES buffer, MES buffer, MOPS buffer, MEM (minimal essential medium), α-MEM (alpha minimal essential medium), DMEM (Dulbecco's modified minimal essential medium), F-12 Medium (Ham's F-12), DMEM / F12 Medium (Dulbecco's Modified Eagle's Medium / Ham's F-12), RPMI 1640 medium, M-199 medium, etc.; to maintain the survival of biological particles, the diluent contains 1-25% serum, with an optimal concentration of 2-10%. The types of serum added include fetal bovine serum (FBS), calf serum (CS), and horse serum (Horse serum). Serum or human serum, etc.; and to prevent biological particles from adhering to the flow channel or carrier, the aforementioned diluent contains anticoagulants, such as ethylenediaminetetraacetic acid (EDTA), heparin, sodium citrate, and potassium oxalate. The diluents used are oxalate and dipotassium ethylenediaminetetraacetate (K2EDTA). The pH and osmotic pressure of the aforementioned diluents are the same as those of the test sample, so that the shape and state of the biological particles remain stable during the detection. The pH of the diluent used to dilute blood samples is generally between pH 6.5 and pH 8.0, with the optimal range between pH 7.0 and pH 7.5, and the osmotic pressure is generally between 200-400 mOsm / kg, with the optimal range between 250-350 mOsm / kg. In addition, the aforementioned diluents are light-transmitting solutions, so that the detection optical path R of the optical detection component 20 can penetrate the flow channel 10 and the diluted test solution, thereby detecting the target biological particles.

[0033] Step S08: Control the flow of the test solution into the flow channel 10 from one inlet end 12. In this embodiment, the flow channel 10 is a microchannel, the inlet end 12 of the flow channel 10 is connected to a sample source 40, and the other end is used to discharge the test solution. Further, a sample supply device 50 can controllably regulate the flow rate of the test solution in the sample source 40 into the flow channel 10, thereby controlling whether the test solution is discharged and controlling the flow rate of the test solution in the flow channel 10.

[0034] Step S10: When the optical detection component 20 senses the target biological particles in the test solution, the carrier disk 30 is controlled to move to an outlet end 14 of the flow channel 10, so that the target biological particles are discharged from the outlet end 14 and collected in the carrier disk 30; wherein the optical detection component 20 includes an excitation light source 22 and a photosensor 24, the light emitted by the excitation light source 22 passes through the flow channel 10 via the detection optical path R and is received by the photosensor 24. In this embodiment, the excitation light source 22 and the light in the optical detection component 20 are... Sensors 24 are disposed on opposite sides of the flow channel 10. In another embodiment, the excitation light source 22 and the photosensor 24 in the optical detection assembly 20 may also be disposed on the same side. The optical detection assembly 20 provides a detection optical path R, which illuminates the target biological particles in the flow channel 10, and the optical detection assembly 20 senses the target biological particles. When the optical detection assembly 20 senses the target biological particles in the test solution, the sample supply device 50 controls the sample source 40 to stop supplying the test solution to the flow channel 10. The flow of the test solution is controlled to stop in the flow channel 10, and the flow channel 10 stops discharging the test solution. When the carrier plate 30 is controlled to move to the outlet end 14 of the flow channel 10, the flow of the test solution in the flow channel 10 is controlled again, so that a portion of the test solution containing the target biological particles is discharged from the outlet end 14 and collected in the carrier plate 30. Subsequently, the carrier plate 30 containing the target biological particles can be moved to a device such as a suction device for subsequent suction of the target biological particles. The above steps can be repeated. Steps S08 and S10 are performed to continue detecting the target biological particles in the test solution. In this embodiment, the carrier disk 30 is moved to the outlet end 14 of the flow channel 10 by a carrier disk unit 60. In this embodiment, the volume of the partial solution discharged into the carrier disk 30 is 5 to 40 μl. Depending on the operator's needs, 5, 10, 15, 20, 25, 30, 35 or 40 μl of partial solution can be discharged into the carrier disk 30. In other embodiments, the volume of the partial solution discharged into the carrier disk 30 can be adjusted according to the operator's needs.

[0035] Furthermore, the excitation light source 22 can be a laser light source, a mercury lamp light source, or an LED light source; the wavelength of the light emitted by the excitation light source 22 can be visible light or invisible light; the photosensor 24 can be a photomultiplier tube (PMT) or a charge-coupled device (CCD); preferably, the optical detection component 20 further includes a filter disposed at the front end of the photosensor 24, and the number of filters can be one or more.

[0036] For example, the excitation light emitted by the excitation light source 22 can enter the flow channel 10 along the detection optical path R. After the target biological particle in the flow channel 10 absorbs the excitation light emitted by the excitation light source 22, it emits a radiation light, which can be fluorescence or cold light. After being filtered by the filter, the wavelength of the radiation light emitted after excitation can pass through the filter and enter the photosensor 24, thereby determining that the optical detection component 20 has sensed the target biological particle.

[0037] Furthermore, in this embodiment, step S10 further includes placing a diluent in the carrier plate 30, controlling the portion of the test solution containing the target biological particles to be discharged from the outlet end 14 and then loaded onto the carrier plate 30 to dilute the portion of the solution; that is, when the portion of the test solution containing the target biological particles is discharged from the outlet end 14 into the carrier plate containing the diluent, the dilution action can be completed simultaneously to facilitate the subsequent absorption of the target biological particles; wherein the diluent can dilute the portion of the solution, making the sample diluted by 50X to 10000X.

[0038] Preferably, the diluent dilutes the partial solution by 10 to 100X, more preferably by 20 to 60X. In this embodiment, the partial solution is 5-40 μl. In one embodiment, the partial solution is 5 μl, and the diluent in the carrier disk is 495 μl, diluting the partial solution by 100X. In another embodiment, the partial solution is 10 μl, and the diluent in the carrier disk is 490 μl, diluting the partial solution by 50X. In another embodiment, the partial solution is 15 μl, and the diluent in the carrier disk is 525 μl, diluting the partial solution by 36X. In yet another embodiment... In one embodiment, the partial solution is 20 μl, and the diluent in the carrier disk is 500 μl, resulting in a 26X dilution of the partial solution; in another embodiment, the partial solution is 25 μl, and the diluent in the carrier disk is 500 μl, resulting in a 21X dilution of the partial solution; in another embodiment, the partial solution is 30 μl, and the diluent in the carrier disk is 570 μl, resulting in a 20X dilution of the partial solution; in another embodiment, the partial solution is 35 μl, and the diluent in the carrier disk is 490 μl, resulting in a 15X dilution of the partial solution; in another embodiment, the partial solution is 40 μl, and the diluent in the carrier disk is 360 μl, resulting in a 10X dilution of the partial solution.

[0039] Furthermore, in this embodiment, step S06 includes diluting the sample by 5X to 100X. In other embodiments, it includes a lower limit of 5X, 10X, or 15X for diluting the sample and an upper limit of 100X, 90X, or 80X for diluting the sample. In this embodiment, step S10 includes diluting the partial solution by 10X to 100X. In other embodiments, it includes a lower limit of 10X, 15X, or 20X for diluting the partial solution and an upper limit of 100X, 80X, or 60X for diluting the partial solution. The lower limit of the total dilution ratio is the lower limit of the sample dilution multiplied by the lower limit of the partial solution dilution, and the upper limit of the total dilution ratio is the upper limit of the sample dilution multiplied by the upper limit of the partial solution dilution.

[0040] In summary, the biological particle screening and detection method of the present invention can perform trace detection. Furthermore, by diluting the sample to form the test solution, when the number of biological particles in the sample is large, it can avoid the influence of clustered biological particles or excessively high concentrations of the sample on the accuracy of the optical detection component 20 in sensing the target biological particles in the test solution, thereby improving the accuracy and stability of trace detection.

[0041] The above description is only a preferred and feasible embodiment of the present invention. Any equivalent changes made by applying the present invention specification and claims should be included within the patent scope of the present invention.

Claims

1. A method for screening and detecting biological particles, comprising: A biological particle screening and detection system is provided, comprising a flow channel, an optical detection component and a carrier disk, wherein the optical detection component provides a detection optical path that penetrates the flow channel, and the optical detection component is used to sense a target biological particle; Provide a sample; The sample was diluted to form a test solution; The test solution is controlled to flow into the flow channel from one inlet end; as well as When the optical detection component senses the target biological particles in the test solution, the carrier disk is controlled to move to an outlet end of the flow channel, wherein a diluent is contained in the carrier disk, and a portion of the test solution containing the target biological particles is discharged from the outlet end and then loaded onto the carrier disk to dilute the portion of the solution.

2. The biological particle screening and detection method as described in claim 1, comprising diluting the sample by 5X to 100X.

3. The biological particle screening and detection method as described in claim 1, comprising mixing the test solution to ensure that the diluted biological particles are uniformly distributed in the solution.

4. The biological particle screening and detection method as described in claim 1, wherein the sample is a blood sample, urine sample, pleural effusion, ascites, spinal fluid, cerebrospinal fluid, or amniotic fluid.

5. The biological particle screening and detection method according to claim 3, wherein the partial solution is diluted to dilute the sample by 50X to 10000X.

6. The biological particle screening and detection method according to claim 3, wherein the diluent dilutes the partial solution to a concentration of 10X to 100X.

7. The biological particle screening and detection method according to claim 3, wherein the diluent dilutes the partial solution to a concentration of 20X to 60X.

8. The biological particle screening and detection method as claimed in claim 1, wherein when the optical detection component senses the target biological particles in the test solution, the test solution is controlled to stop flowing in the flow channel; when the carrier is controlled to move to the outlet end of the flow channel, the test solution is controlled to flow in the flow channel again, so that a portion of the solution containing the target biological particles in the test solution is discharged from the outlet end and loaded onto the carrier.

9. The biological particle screening and detection method as described in claim 1, wherein the optical detection component comprises an excitation light source and a photosensor, and the light emitted by the excitation light source passes through the flow channel via the detection optical path and is received by the photosensor.

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