Physical test system and test method of single cell

Abstract

The invention provides a single cell physical test system and test method, and relates to the technical field of biological test. The physical test system comprises an illumination assembly, a carrying mechanism, an optical tweezers generation assembly, an optical path coupling assembly, an optical splitter, an imaging assembly and a spectrograph. Wherein the illumination assembly generates illumination light; the loading mechanism comprises a displacement platform and a sample table, the sample table is located on the first light path, and the displacement platform drives the sample table to move; the optical tweezer generation assembly generates optical tweezer light; the light path coupling assembly guides the illumination light to be emitted and guides the optical tweezer light to irradiate the sample table along a second light path; the optical splitter is used for splitting the illumination light into first signal light and second signal light; the imaging assembly couples and receives the first signal light; the spectrometer couples and receives the second signal light. According to the physical test system, the optical tweezers and the spectrum are combined, and morphology imaging, spectrum testing and mechanical property testing of single cells can be achieved at the same time only by adjusting and controlling the position of the sample table.

Description

Technical Field

[0001] The present disclosure relates to the technical field of biological testing, and particularly to a physical testing system and method for single cells. Background Art

[0002] The characteristics and behaviors of single cells can often characterize the overall state of a biological system. The heterogeneity among individual cells plays a crucial role in the processes of disease development, diagnosis, and treatment with drugs. By measuring the morphology, structure, and force-related properties of single cells, it is possible to provide a basis for research on disease diagnosis, pathology, cell development and differentiation, drug R & D and screening, and immune responses, etc.

[0003] Currently, flow cytometers and Raman spectrometers are mainly used clinically to detect the physical properties of single cells. However, such detection devices and methods all have the disadvantages of high cost, complex operation, and lack of simultaneous measurement of the cell structure, morphology, and related forces. For example, the testing process of using a flow cytometer in combination with an optical microscope to lock and reposition a single cell is too complex and requires trained personnel to perform. Moreover, the instrument is expensive and has high requirements for the activity, concentration, etc. of the sample. Although atomic force microscopes (AFMs) and cell traction microscopes (CTMs) can detect the mechanical properties of cells, they also have limitations such as complex operation and high cost.

[0004] Therefore, there is a need to provide a physical testing system and method for single cells to improve the above problems. Summary of the Invention

[0005] The purpose of the present disclosure is to provide a physical testing system and method for single cells to improve the technical problem that current cell detection instruments cannot perform spectral and mechanical tests on single cells in a convenient operation manner.

[0006] To achieve the above object and other related objects, in a first aspect, the present disclosure provides a physical testing system for single cells, which includes an illumination component, a sample stage mechanism, an optical tweezer generation component, an optical path coupling component, a beam splitter, an imaging component, and a spectrometer.

[0007] Wherein, the illumination component generates illumination light transmitted along the first optical path; the sample-carrying mechanism includes a displacement platform and a sample stage disposed on the displacement platform, the sample stage is located on the first optical path, and the displacement platform drives the sample stage to move; the optical tweezer generating component is used to generate optical tweezer light; the optical path coupling component guides the illumination light to exit, and guides the optical tweezer light to irradiate the sample stage along the second optical path, the optical tweezer light forms an optical trap on the sample stage, and the transmission direction of the second optical path coincides with that of the first optical path in an opposite manner; the beam splitter is located on the light-emitting side where the optical path coupling component transmits the illumination light, and the beam splitter is used to split the illumination light into a first signal light and a second signal light; the imaging component coupledly receives the first signal light and converts the first signal light into a detection image; the spectrometer coupledly receives the second signal light.

[0008] In an example of the present invention, the optical path coupling component includes a dichroic mirror, and the dichroic mirror is coupled with the first optical path and corresponds to the light-emitting side of the optical tweezer generating component.

[0009] In an example of the present invention, an objective lens and a tube lens are provided between the optical path coupling component and the sample stage, and the objective lens and the tube lens are sequentially arranged along the transmission direction of the first optical path.

[0010] In an example of the present invention, the spectrometer receives the second signal light through a first optical fiber, the end of the first optical fiber has an optical fiber coupler, the optical fiber coupler is used to coupledly receive the second signal light, and a diaphragm is provided on the optical fiber coupler.

[0011] In an example of the present invention, the illumination component includes an illumination light source and a condenser lens, the illumination light source generates the illumination light, and the condenser lens is located on the first optical path and is disposed between the illumination light source and the sample stage.

[0012] In an example of the present invention, the wavelength of the illumination light is 350nm - 700nm.

[0013] In an example of the present invention, the optical tweezer generating component includes a modulation light source and an optical modulator, the modulation light source generates coherent light, and the optical modulator modulates the coherent light into the optical tweezer light.

[0014] In a second aspect, the present disclosure provides a physical testing method for single cells. This physical testing method is implemented using the physical testing system described in any one of the above examples. This physical testing method includes: Placing a cell sample on the sample stage, the cell sample includes a physiological saline solution and cells placed in the physiological saline solution; Turn on the illumination component and the optical tweezer generation component, so that the illumination light and the optical tweezer light irradiate the sample stage in opposite directions, and the optical tweezer light forms an optical trap on the sample stage; Use the imaging component to convert the first signal light into a detection image, and the detection image shows the image of the sample stage covered by the illumination light; Drive the sample stage to move through the displacement platform, and drive the single cell to be measured in the cell sample to move to the middle position of the detection image, so that the optical trap captures and fixes the single cell to be measured; Use the spectrometer to receive the second signal light, and obtain the spectral data of the single cell to be measured based on the second signal light.

[0015] In an example of the present invention, obtaining the spectral data of the single cell to be measured based on the second signal light includes: Convert the second signal light into a test spectrum; based on the test spectrum and the dark current spectrum and background spectrum pre-measured by the spectrometer, determine the absorption spectrum of the single cell to be measured.

[0016] In an example of the present invention, the physical test method further includes: Drive the sample stage to move at a preset speed through the displacement platform, so that the single cell to be measured deforms; based on the deformation ratio of the single cell to be measured, the preset speed, and the viscosity coefficient of the physiological saline solution, determine the elastic modulus of the single cell to be measured; wherein, the deformation ratio is the ratio of the deformation size of the single cell to be measured to the original size of the single cell to be measured.

[0017] The single cell physical test system provided by the present invention combines optical tweezers with illumination light, and only by adjusting the position of the sample stage, the morphological imaging, spectral testing and mechanical property testing of single cells can be realized simultaneously. The physical test system can complete the characterization of the metabolic situation of single cells through spectral testing, and can also realize the quantitative analysis of the elastic modulus, tensile recovery ability and deformation behavior in the shear force field of the tested single cell under different environments, so as to more comprehensively reveal the influence of different action conditions on the cell metabolic situation through the two dimensions of coupling dynamics - metabolism. Description of the Drawings

[0018] The features and advantages of the present disclosure will be more clearly understood by referring to the accompanying drawings. The drawings are schematic and should not be construed as imposing any limitation on the present disclosure. In the drawings: Figure 1 Shows a schematic structural diagram of a physical test system in an embodiment of the present disclosure; Figure 2 Shows a schematic structural diagram of a physical test system in an embodiment of the present disclosure; Figure 3It is a schematic flow chart of a physical test method in an embodiment of the present disclosure; Figure 4 It is a schematic flow chart of step S5 in an embodiment of the present disclosure; Figure 5 It is a schematic flow chart of step S6 in an embodiment of the present disclosure; Figure 6 It is a transmission spectral image of a single red blood cell measured by a physical test system in an embodiment of the present disclosure; Figure 7 It is an absorption spectrum of a single red blood cell measured by a physical test system in an embodiment of the present disclosure; Figure 8 It is an absorption spectrum simulating different proportions of oxyhemoglobin in an embodiment of the present disclosure; Figure 9 It shows that in an embodiment of the present disclosure, the peak value of the β band / the peak value of the α band of the simulated standard spectrum is correlated with the oxygenation ratio ω; Figure 10 It is a diagram of the stretching process of a single red blood cell measured by a physical test system in an embodiment of the present disclosure.

[0019] Element reference numeral description: 10. Lighting assembly; 11. Lighting light source; 12. Condensing lens; 20. Sample stage; 21. Objective lens; 22. Tube lens; 30. Optical path coupling assembly; 31. Dichroic mirror; 32. Reflecting mirror; 40. Optical tweezer generating assembly; 41. Modulating light source; 42. Second optical fiber; 43. Collimator; 44. Beam expander; 45. Transmissive and reflective mirror; 46. Optical modulator; 50. Spectrometer; 61. Camera; 62. Image processor; 70. Spectrometer; 71. First optical fiber; 72. Fiber optic coupler; 73. Aperture; 80. Optical tweezer - spectrum cooperation module; 81. Housing; 82. Light - passing hole; A. First optical path; B. Second optical path. Detailed implementation manners

[0020] Next, the technical solutions in the embodiments of the present disclosure will be clearly and completely described in conjunction with the accompanying drawings in the embodiments of the present disclosure. Obviously, the described embodiments are part of the embodiments of the present disclosure, rather than all the embodiments. Based on the embodiments in the present disclosure, all other embodiments obtained by those of ordinary skill in the art without making creative efforts belong to the scope of protection of the present disclosure.

[0021] Please refer to Figures 1 to 10It should be noted that the illustrations provided in this embodiment only schematically illustrate the basic concept of the present disclosure. Therefore, only the components related to the present disclosure are shown in the drawings, rather than being drawn according to the number, shape, and size of the components in actual implementation. The type, quantity, and ratio of each component in actual implementation can be arbitrarily changed, and the component layout type may also be more complex.

[0022] Please refer to Figure 1 and Figure 2 , in a first aspect, the present disclosure provides a physical testing system for single cells. This physical testing system provides a single-cell optical tweezers-spectroscopy integrated testing and analysis system with low design cost and convenient operation. After detecting the spectrum of a single cell through a microscopic system, this physical testing system can use optical tweezers to measure and operate the single cell to test and obtain mechanical parameters such as the adhesion force and elastic modulus of the single cell. Compared with flow cytometers, Raman spectroscopy, etc., this physical testing system has a lower cost and does not require complex operations, and can detect more abundant indicators.

[0023] As Figure 1 and Figure 2 shown, this physical testing system includes an illumination component 10, a sample stage mechanism, an optical tweezers generation component 40, an optical path coupling component 30, a beam splitter 50, an imaging component, and a spectrometer 70.

[0024] The illumination component 10 is capable of generating illumination light that travels along a first optical path A. This illumination light irradiates the cell sample to perform imaging and spectral analysis on the cell sample. The wavelength of this illumination light can be set according to the spectral analysis requirements. For example, the wavelength of the illumination light can be 350 nm - 700 nm.

[0025] The sample stage mechanism includes a displacement platform and a sample stage 20. The sample stage 20 is arranged on the displacement platform. The sample stage 20 is located on the light output side of the illumination component 10 and is arranged on the first optical path A. The displacement platform is a three-axis piezoelectric displacement platform and a two-dimensional platform. The two-dimensional platform can realize the field of view switching of different sample stages 20. The three-axis piezoelectric displacement platform can drive the sample stage 20 to displace in three-dimensional space. The displacement accuracy of the three-axis piezoelectric displacement platform can reach the nanometer level, and it can realize the automatic focusing of the illumination light on the sample stage 20 through program control, and can also realize the two-dimensional movement of the sample stage 20 in the horizontal direction.

[0026] The optical tweezer generating component 40 is used to generate optical tweezer light, which can form an optical trap on the imaging plane through spatial modulation. The optical path coupling component 30 is simultaneously coupled to the light output side of the optical tweezer generating component 40 and the light output side of the sample stage 20 for transmitting illumination light. The optical path coupling component 30 is used to guide the illumination light to exit and guide the optical tweezer light to irradiate the sample stage 20 along the second optical path B. The transmission direction of the second optical path B coincides with that of the first optical path A in the opposite direction, so that the optical tweezer light and the illumination light are combined and transmitted in the reverse direction and coincide on the sample stage 20. The optical tweezer light forms an optical trap on the sample stage 20 to capture and fix the cells on the sample stage 20.

[0027] The beam splitter 50 is located on the light output side of the optical path coupling component 30 for transmitting illumination light. The beam splitter 50 splits the incident illumination light into a first signal light and a second signal light, and the intensities of the first signal light and the second signal light are the same. As Figure 6 shown, the imaging component is coupled to receive the first signal light and converts the first signal light into a detection image, which is an image showing the area of the sample stage 20 covered by the illumination light, so as to complete the image detection of the morphology and structure of the single cell to be measured. As Figure 7 shown, the spectrometer 70 is coupled to receive the second signal light and performs spectral analysis using the second signal light to obtain the spectral data of the single cell to be measured. The spectral data can be used for test spectra such as absorption spectra and Raman spectra.

[0028] As Figure 2 shown, in some embodiments, the optical path coupling component 30 includes a dichroic mirror 31. The dichroic mirror 31 is coupled to the first optical path A and corresponds to the light output side of the optical tweezer generating component 40. The dichroic mirror 31 can transmit the illumination light by being coupled to the first optical path A. The dichroic mirror 31 can also receive the incident optical tweezer light and reflect and guide the optical tweezer light to be combined with the illumination light and transmitted in the reverse direction (i.e., transmitted along the second optical path B). For example, in an example, the optical path coupling component 30 includes a reflecting mirror 32 and a dichroic mirror 31. The reflecting mirror 32 is arranged on the first optical path A, and the reflecting surface of the reflecting mirror 32 faces the light output side of the sample stage 20 for transmitting illumination light; the dichroic mirror 31 corresponds to the reflecting surface of the reflecting mirror 32 and the light output side of the optical tweezer generating component 40. On the one hand, the dichroic mirror 31 can transmit the illumination light reflected by the reflecting mirror 32, and on the other hand, the dichroic mirror 31 can completely reflect the optical tweezer light, so that the optical tweezer light and the illumination light are incident on the reflecting mirror 32 in the same beam and in the reverse direction, and then are reflected by the reflecting mirror 32 and irradiate the sample stage 20 along the second optical path B.

[0029] As Figure 1 and Figure 2As shown, in some embodiments, an objective lens 21 and a tube lens 22 are further provided between the sample stage 20 and the optical path coupling assembly 30, and the objective lens 21 and the tube lens 22 are arranged in sequence along the transmission direction of the first optical path A. Among them, the objective lens 21 performs inverted microscope imaging on the illumination light coverage area on the sample stage 20. The front pupil of the objective lens 21 faces the sample stage 20, and the rear pupil of the objective lens 21 faces away from the sample stage 20. The tube lens 22 is arranged on one side of the rear pupil of the objective lens 21. It should be noted that the objective lens 21 can be replaced with various series of objective lenses 21 with a magnification of 4X - 100X according to requirements. For example, in one example, the objective lens 21 is a water lens, and the NA value of the objective lens 21 is 1.2 and the magnification is 60X.

[0030] As Figure 1 and Figure 2 As shown, in some embodiments, a first optical fiber 71 is connected to the spectrometer 70, and the end of the first optical fiber 71 has an optical fiber coupler 72. The optical fiber coupler 72 is used to couple and receive the first signal light, so that the first signal light is output to the spectrometer 70 along the first optical fiber 71. Among them, a diaphragm 73 is provided on the optical fiber coupler 72, and the aperture of the diaphragm 73 can be adaptively adjusted according to the size of the single cell to be measured and the magnification of the objective lens 21, so that the aperture of the diaphragm 73 is slightly larger than the size of the single cell magnified by the objective lens 21, thereby ensuring that the spectrometer 70 only receives the transmitted light of the single cell to be measured and filters the background light outside the single cell to be measured, so as to avoid serious noise interference of the background light on the spectral data.

[0031] As Figure 1 and Figure 2 As shown, in some embodiments, the imaging assembly includes an imager 61 and an image processor 62. The imager 61 is used to couple and receive the first signal light, and the image processor 62 is communicatively connected to the imager 61. The image processor 62 can convert the first signal light into a detection image. In addition, in some embodiments, the image processor 62 can also be communicatively connected to the spectrometer 70 and convert the spectral data generated by the spectrometer 70 into a spectral image.

[0032] As Figure 1 and Figure 2 As shown, in some embodiments, the illumination assembly 10 includes an illumination light source 11 and a condenser lens 12. The illumination light source 11 generates illumination light. The illumination light source 11 can adopt a broadband light source with a stable intensity. For example, the illumination light source 11 can adopt a halogen lamp. The condenser lens 12 is arranged on the first optical path A and is located between the illumination light source 11 and the sample stage 20. The condenser lens 12 focuses the illumination light on the sample stage 20. The condenser lens 12 can uniformly irradiate the illumination light on the sample stage 20 to achieve Köhler illumination of the illumination light on the sample stage 20, and the illumination light is uniformly distributed and has a consistent light intensity on the sample stage 20.

[0033] As Figure 1 and Figure 2As shown, the optical tweezer generation component 40 includes a modulated light source 41 and an optical modulator 46. The modulated light source 41 generates coherent light. For example, the modulated light source 41 uses a laser with an output wavelength of 1064 nm and a power range of 200 mW - 5 W. The optical modulator 46 is disposed on the light output side of the modulated light source 41 and modulates the coherent light into optical tweezer light. The optical modulator 46 can control and adjust the position and number of optical traps formed by the optical tweezer light on the imaging surface. The optical modulator 46 is an acousto-optic deflector or a spatial light modulator 46. As Figure 2 As shown, in one example, the optical tweezer generation component 40 includes a modulated light source 41, a collimator 43, a beam expander 44, a dichroic mirror 45, and an optical modulator 46. The modulated light source 41 outputs coherent light through a second optical fiber 42. A collimator 43 is coupled to the light output end of the second optical fiber 42, and the collimator 43 adjusts the output coherent light into parallel light. The beam expander 44 is disposed on the light output side of the collimator 43, and the beam expander 44 expands the coherent light for output. The dichroic mirror 45 and the optical modulator 46 are sequentially disposed on the light output side of the beam expander 44 along the coherent light output direction. The light transmission surface of the dichroic mirror 45 corresponds to the light output side of the beam expander 44, and the light reflecting surface of the dichroic mirror 45 corresponds to the optical modulator 46. The dichroic mirror 45 transmits the coherent light so that the coherent light is incident on the optical modulator 46. After the optical modulator 46 modulates the coherent light into optical tweezer light, the optical tweezer light is reflected to the light reflecting surface of the dichroic mirror 45. The dichroic mirror reflects the optical tweezer light and collimates it through a 4f lens system and then incident on the optical path coupling component 30, and finally is guided by the optical path coupling component 30 and irradiated on the sample stage 20 along the second optical path B.

[0034] In addition, as Figure 1 and Figure 2 As shown, in some embodiments, the optical path coupling component 30, the beam splitter 50, the imaging component, the receiving end of the spectrometer 70, and the optical tweezer generation component 40 except the modulated light source 41 are integrated into a housing 81 to form an optical tweezer - spectroscopy synergy module 80. A light passing hole 82, coupling ports for a first optical fiber 71 and a second optical fiber 42, and a data line port of the imaging component are provided on the housing 81 of the optical tweezer - spectroscopy synergy module 80. The light passing hole 82 is a port for receiving illumination light into the optical path coupling component 30 and outputting optical tweezer light.

[0035] Please refer to Figures 3 to 5 , in a second aspect, the present disclosure provides a physical testing method for a single cell. This physical testing method is implemented using the physical testing system described in any one of the above embodiments. This physical testing method includes the following steps: S1. Place the cell sample on the sample stage 20. The cell sample includes a physiological saline solution and cells placed in the physiological saline solution.

[0036] In step S1, a cell sample is first prepared and uniformly coated on the sample stage 20. When detecting red blood cells, for example, the blood sample is mixed with 1xPBS solution at a certain ratio, the dilution factor is recorded, and the cell sample liquid is added to the sample stage 20 with a depth of about 100 μm.

[0037] Next, step S2 is executed: the illumination component 10 and the optical tweezer generation component 40 are turned on, so that the illumination light and the optical tweezer light are irradiated onto the sample stage 20 in opposite directions, and the optical tweezer light forms an optical trap on the sample stage 20.

[0038] In step S2, the illumination light source 11 is turned on, and the condenser 12 is adjusted so that Kohler illumination is achieved on the sample stage 20; the optical tweezer generation component 40 is turned on, and the optical tweezer light is generated by the optical tweezer generation component 40. The optical tweezer light is irradiated onto the sample stage 20 through the optical path guidance and forms an optical trap on the sample stage 20.

[0039] Next, step S3 is executed: the imaging component is used to convert the first signal light into a detection image, and the detection image shows the image of the sample stage 20 covered by the illumination light.

[0040] In step S3, the imaging component is turned on, and the imaging component is used to receive the first signal light formed by the transmission of the illumination light and form a detection image showing the area of the sample stage 20 irradiated and covered by the illumination light; at the same time, by adjusting the axial position of the objective lens 21, the imaging component can clearly display the detection image, and then clearly display the cell image in the cell sample.

[0041] Next, step S4 is executed: the sample stage 20 is driven to move by the displacement platform, and the single cell to be measured in the cell sample is driven to move to the middle position of the detection image, so that the optical trap captures and fixes the single cell to be measured.

[0042] As Figure 6 shown, in step S4, the sample stage 20 is driven to move horizontally by controlling the displacement platform, and the single cell to be measured in the cell sample is driven to move to the overlapping area of the illumination light and the optical tweezer light on the sample stage 20 (i.e., the middle position of the detection image), so as to facilitate capturing the single cell to be measured and detecting the spectral parameters and mechanical parameters of the single cell to be measured in the subsequent process.

[0043] Next, step S5 is executed: the spectrometer 70 is used to receive the second signal light, and the spectral data of the single cell to be measured is obtained based on the second signal light.

[0044] As Figure 4 and Figure 7 shown, the spectral data includes the absorption spectrum of the single cell to be measured. In some embodiments, the process of obtaining the absorption spectrum of the single cell to be measured in step S5 includes the following steps: S51. Convert the second signal light into a test spectrum.

[0045] In step S51, the spectrometer 70 converts the second signal light into a test spectrum.

[0046] S52. Based on the test spectrum and the dark current spectrum and background spectrum pre-measured by the spectrometer 70, determine the absorption spectrum of the single cell to be measured.

[0047] In step S52, filter the noise effects of the dark current spectrum and the background spectrum in the test spectrum to obtain the absorption spectrum of the single cell to be measured.

[0048] As Figure 8 and Figure 9 shown, for the absorption spectrum, according to the standard spectral database, the molar extinction coefficient curves of the ratio of oxyhemoglobin (HbO2) and hemoglobin (Hb) in the 360 - 700 nm band can be simulated. It can be known that the β-band peak / α-band peak in the Q-band (500 - 600 nm) is correlated with the oxygenation ratio ω, and based on this, the linear response interval of the oxygenation ratio ω can be determined.

[0049] As Figure 7 shown, by performing step S5 to obtain the absorption spectrum of the single cell to be measured within the above-determined 360 - 700 nm band, and evaluate the hemoglobin content of the cell by calculating the β-band peak / α-band peak in the absorption spectrum.

[0050] In addition, as Figure 5 and Figure 10 shown, in some embodiments, the physical test method further includes an elastic modulus test step S6 for the single cell to be measured. The elastic modulus test step S6 includes: S61. Drive the sample stage 20 to move at a preset speed through the displacement stage, and drag the single cell to be measured fixed by optical trapping, so that the single cell to be measured deforms.

[0051] S62. Based on the deformation ratio of the single cell to be measured, the preset speed of the displacement stage movement, and the viscosity coefficient of the solution, determine the elastic modulus of the single cell to be measured.

[0052] As Figure 10 shown, when the single cell to be measured is a red blood cell, since the red blood cell is biconcave disc-shaped, when the red blood cell flows in an infinite space, the optical trapping force F t acting on the trapped red blood cell F d balances with the fluid resistance F d At this time, the fluid resistance (1) In formula (1), η is the viscosity coefficient of the solution in the cell sample, d 0 is the initial diameter of the single cell to be measured when no deformation occurs, v is the relative velocity between the single cell to be measured and the solution fluid (i.e., the preset velocity of the displacement platform movement). Since the elastic modulus of the cell is linear when the single cell to be measured is stretched, the stretching force of the solution fluid can be related to the cell elongation. In step S62, based on the linear relationship between the stretching force of the solution fluid and the cell deformation, according to formula (2), based on the deformation ratio λ of the single cell to be measured, the preset velocity v of the displacement platform movement, η and the viscosity coefficient μ of the solution, the elastic modulus (2) of the single cell to be measured is determined. Formula (2) is as follows: λ In formula (2), λ is the deformation ratio of the single cell to be measured, and the deformation ratio λ is expressed as d =Δ d / d 0, where Δ d s - d 0, d s is the diameter of the single cell to be measured on the major axis after being stretched and deformed, d 0 is the initial diameter of the single cell to be measured when not stretched.

[0053] In summary, the single cell physical testing system provided by the present invention combines an optical tweezer with illumination light. By only regulating the position of the sample stage, the morphological imaging, spectral testing, and mechanical property testing of a single cell can be simultaneously achieved. While the physical testing system can complete the characterization of the metabolic condition of a single cell through spectral testing, it can also quantitatively analyze the elastic modulus, stretching recovery ability, and deformation behavior in a shear force field of the tested single cell under different environments. Thus, through the two dimensions of kinetics - metabolism, the influence of different action conditions on the cell metabolic condition can be more comprehensively revealed.

[0054] The above embodiments are only illustrative of the principles and effects of the present invention, and are not used to limit the present invention. Any person familiar with this technology can modify or change the above embodiments without departing from the spirit and scope of the present invention. Therefore, all equivalent modifications or changes completed by those with ordinary knowledge in the technical field without departing from the spirit and technical ideas disclosed by the present invention should still be covered by the claims of the present invention.

Claims

1. A single-cell physical testing system, characterized in that, Comprising: A lighting component that generates illumination light transmitted along a first optical path; A sample-carrying mechanism that includes a displacement platform and a sample stage disposed on the displacement platform. The sample stage is located on the first optical path, and the displacement platform drives the sample stage to move; An optical tweezer generation component for generating optical tweezer light; An optical path coupling component that guides the illumination light to exit and guides the optical tweezer light to irradiate the sample stage along a second optical path. The optical tweezer light forms an optical trap on the sample stage, and the transmission direction of the second optical path coincides with that of the first optical path in the opposite direction; A beam splitter located on the light-emitting side where the optical path coupling component transmits the illumination light. The beam splitter is used to split the illumination light into a first signal light and a second signal light; An imaging component that coupledly receives the first signal light and converts the first signal light into a detection image; A spectrometer that coupledly receives the second signal light.

2. The physical test system according to claim 1, wherein The optical path coupling component includes a dichroic mirror that is coupled to the first optical path and corresponds to the light-emitting side of the optical tweezer generation component.

3. The physical test system according to claim 1, wherein An objective lens and a tube lens are disposed between the optical path coupling component and the sample stage, and the objective lens and the tube lens are sequentially arranged along the transmission direction of the first optical path.

4. The physical test system according to claim 3, wherein The spectrometer receives the second signal light through a first optical fiber. The end of the first optical fiber has an optical fiber coupler for coupledly receiving the second signal light, and a diaphragm is disposed on the optical fiber coupler.

5. The physical test system according to claim 1, wherein The lighting component includes a lighting source and a condenser lens. The lighting source generates the illumination light, and the condenser lens is located on the first optical path and is disposed between the lighting source and the sample stage.

6. The physical test system according to claim 1 or 5, characterized in that, The wavelength of the illumination light is 350nm - 700nm.

7. The physical test system according to claim 1, wherein, The optical tweezer generation component includes a modulation light source and an optical modulator. The modulation light source generates coherent light, and the optical modulator modulates the coherent light into the optical tweezer light.

8. A physical testing method for single cells, characterized in that, The physical testing method is implemented using the physical testing system according to any one of claims 1 to 7. The physical testing method includes: Placing a cell sample on the sample stage. The cell sample includes a physiological saline solution and cells placed in the physiological saline solution; Turning on the lighting component and the optical tweezer generation component so that the illumination light and the optical tweezer light irradiate the sample stage in opposite directions, and the optical tweezer light forms an optical trap on the sample stage; Using the imaging component to convert the first signal light into a detection image, and the detection image shows an image of the sample stage covered by the illumination light; Driving the sample stage to move through the displacement platform, driving the single cell to be measured in the cell sample to move to the middle position of the detection image, so that the optical trap captures and fixes the single cell to be measured; Using the spectrometer to receive the second signal light and obtaining spectral data of the single cell to be measured based on the second signal light.

9. The physical test method according to claim 8, wherein Further comprising: Driving the sample stage to move at a preset speed through the displacement platform so that the single cell to be measured is deformed; Determine the elastic modulus of the single cell to be measured based on the deformation ratio of the single cell to be measured, the preset speed, and the viscosity coefficient of the physiological saline solution; wherein, the deformation ratio is the ratio of the deformation size of the single cell to be measured to the original size of the single cell to be measured.

10. The physical testing method according to claim 8, wherein The obtaining the spectral data of the single cell to be measured based on the second signal light includes: Convert the second signal light into a test spectrum; Determine the absorption spectrum of the single cell to be measured based on the test spectrum, the dark current spectrum, and the background spectrum pre-measured by the spectrometer.

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