Low-energy laser-assisted cell culture and online monitoring device

Abstract

The present invention relates to the field of cell culture technology, and in particular to a low-energy laser-assisted cell culture and online monitoring device, comprising a cell culture box, a control center, a light source body and a functional body; wherein the cells to be cultured are placed in the functional body, and the control center controls the light source body to emit LED light and all low-energy lasers to the functional body, and the LED light and all low-energy lasers simultaneously complete the illumination and radiation culture of the cells to be cultured; the functional body photographs the cells to be cultured, and transmits the photographed cell images to the control center. The device provided by the present invention can automatically use low-energy lasers of multiple bands in a cell culture box to irradiate cells and monitor cells online during the entire growth process, and can complete long-term culture, irradiation and observation of cells without manually taking out the cells, avoiding changes in the cell growth environment, thereby facilitating the exploration of optimal culture conditions, improving the success rate of cell culture, and reducing the complexity of experiments.

Description

Technical Field

[0001] The present invention belongs to the technical field of cell culture, and in particular relates to a low-energy laser-assisted cell culture and online monitoring device. Background Art

[0002] Stem cells are multipotent cells with the ability to self-replicate. Under certain conditions, they can differentiate into a variety of functional cells. Regenerative medicine, based on the repair and regenerative abilities of stem cells, has the potential to address major medical challenges facing humanity, helping us realize our dream of repairing traumatic and pathological tissues and curing terminal illnesses, potentially triggering a new revolution in medical technology following the success of medication and surgery. Low-energy lasers can enhance the biological activity of stem cells, promote cell proliferation, increase the activity of cellular antioxidant enzymes, strengthen their resistance to the external environment, stimulate the secretion of various cytokines, regulate the wound healing process, and promote wound healing and recovery. They are considered a key factor in tissue engineering.

[0003] Current low-energy laser irradiation therapies lack real-time imaging capabilities within the incubator, typically requiring removal of the incubator for observation under a phase-contrast microscope. Current incubator online monitoring systems are difficult to use with low-energy lasers due to light source limitations. During laser-assisted culture, how to simultaneously monitor the morphological characteristics of stem cells at each stage online, thereby guiding the optimization and improvement of in vitro stem cell culture systems, has become an urgent challenge. Therefore, the development of a low-energy laser stem cell-assisted culture system with online monitoring capabilities is a future development trend and is of great significance for promoting stem cell photobiomodulation therapy. Summary of the Invention

[0004] In view of this, the present invention aims to provide a low-energy laser-assisted cell culture and online monitoring device, which couples multi-wavelength low-energy laser with LED light, and realizes irradiation culture of the cells to be cultured while illuminating and photographing them. Long-term culture, irradiation and observation of cells can be completed in the box without manually removing the cells, avoiding changes in the cell growth environment, thereby facilitating the exploration of optimal culture conditions, improving the success rate of cell culture, and reducing the complexity of the experiment.

[0005] To achieve the above object, the technical solution created by the present invention is implemented as follows:

[0006] A laser-assisted cell culture device comprises a light source body (1), a functional body (2) and a cell culture box (3), wherein the functional body (2) is arranged inside the cell culture box (3), and the light source body (1) is arranged outside the cell culture box (3); wherein cells to be cultured are placed in the functional body (2), and the light source body (1) transmits LED light for illuminating the cells to be cultured and low-energy laser light for irradiating the cells to be cultured to the functional body (2).

[0007] Furthermore, the light source body (1) includes a laser modulation module (4), an LED light source (5) and an optical fiber coupler (6); the laser modulation module (4) emits at least two beams of low-energy laser light for irradiating the cells to be cultured, and the wavelength of each beam of low-energy laser light is different; the LED light source (5) emits LED light for illuminating the cells to be cultured; the optical fiber coupler (6) couples the LED light and all the low-energy laser light into the optical fiber; and the optical fiber transmits the coupled light beams to the functional body (2).

[0008] Furthermore, the laser modulation module (4) includes no less than two lasers (8), each laser (8) emits low-energy laser light, and the wavelength of each low-energy laser light is different.

[0009] Furthermore, the light source body (1) further comprises a light source chassis (7), and the laser modulation module (4), the LED light source (5) and the optical fiber coupler (6) are arranged in the light source chassis (7).

[0010] Furthermore, the functional body (2) includes a point light source emitter (11), a stage (12), a plane reflector (13) and a collimating lens (14); wherein the cells to be cultured are placed on the stage (12); the point light source emitter (11) receives the coupled light beam and irradiates the cells to be cultured with the coupled light beam in the form of a point light source; the light emitted by the point light source emitter (11) is reflected by the plane reflector (13) onto the collimating lens (14); the collimating lens (14) converts the reflected light into a parallel light beam; and the parallel light beam is irradiated onto the cells to be cultured.

[0011] Furthermore, the functional body (2) further includes a functional chassis (10), a point light source emitter (11), a stage (12), a plane reflector (13) and a collimating lens (14) are arranged in the functional chassis (10), and the surface of the functional chassis (10) is sealed.

[0012] Furthermore, a reserved hole is provided on the side wall of the cell culture chamber (3), so that the optical fiber (9) extends into the cell culture chamber (3) through the reserved hole, and the optical fiber coupler (6) couples the LED light and all low-energy lasers into the optical fiber (9). The other end of the optical fiber (9) is connected to the point light source emitter (11), and the point light source emitter (11) emits the coupled light beam in the form of a point light source.

[0013] A laser-assisted cell culture and online monitoring device, comprising the laser-assisted cell culture device provided by the present invention, and also comprising a cell online monitoring device and a control center (19); wherein,

[0014] The cell online monitoring device comprises an objective lens (15), a camera (16) and an electric displacement stage (17); wherein light reflected by the cells to be cultured enters the camera (16) through the objective lens (15), so that the cells to be cultured are imaged on the target surface of the camera (16); the camera (16) is placed on the electric displacement stage (17); the electric displacement stage (17) changes the distance between the camera (16) and the cells to be cultured, thereby completing the focusing of the camera (16);

[0015] The control center (19) controls the laser modulation module (4) so ​​that each laser (8) in the laser modulation module (4) emits low-energy laser light; the control center (19) controls the LED light source (5) to emit LED light; the control center (19) is connected to the camera (16) so that the camera (16) transmits the captured cell image to the control center (19); the control center (19) controls the electric displacement stage (17) so that the electric displacement stage (17) changes the distance between the camera (16) and the cells to be cultured, thereby completing the focusing of the camera (16).

[0016] Furthermore, an attenuation plate and a quarter-wave plate are plated at the center of the rear focal plane of the objective lens (15), so that the parallel light beams form phase contrast interference after passing through the objective lens (15).

[0017] Furthermore, the control center (19) has a timing function and is equipped with a setting program. The setting program is used to set the wavelength, energy density, duration, interval and frequency of laser irradiation and cell photography, so that the control center (19) can automatically control the laser modulation module (4), LED light source (5) and camera (16) to perform the set tasks in a timely manner; the control center (19) has a data analysis function, analyzes the cell image, and draws a laser-cell growth curve in combination with the current laser control data.

[0018] Compared with the prior art, the present invention can achieve the following beneficial effects:

[0019] (1) The laser-assisted cell culture device created by the present invention can be equipped with a variety of low-energy laser light sources of different wavelengths as needed to achieve irradiation culture of cells to be cultured; equipped with an LED light source for imaging to avoid the laser speckle effect during imaging; and multiple low-energy lasers are coupled with LED light and then transmitted to the functional body, thereby achieving low-energy laser irradiation of cells to be cultured and cell status monitoring, eliminating the interference of the light source on the temperature in the cell culture box, and being able to perform low-energy laser irradiation experiments on cells without opening the functional body;

[0020] (2) The laser-assisted cell culture device created by the present invention separates the functional body and the light source body. The functional body only retains the necessary components in a conventional cell culture box, and selects a small and compact model to reduce the internal space occupied by the incubator. The functional body can be placed in the incubator at will, and the light source body is placed outside the box. This design can directly empower the existing cell culture box and realize the functions of low-energy laser irradiation and cell status monitoring in the box without replacing the existing equipment, avoiding the tedious operation of opening the incubator to remove cells in the prior art, thereby ensuring the constancy of the cell growth environment and improving the success rate of experimental operations.

[0021] (3) The laser-assisted cell culture device created by the present invention converts the light emitted by the point light source emitter into parallel light by providing a collimating lens, thereby effectively improving the image contrast and flat field of the captured image, so that the imaging quality at the edge of the image is equivalent to that at the center of the image;

[0022] (4) The control center of the low-energy laser-assisted cell culture and online monitoring device created by the present invention is connected to a camera. The operator can observe the growth status of cells in the cell culture incubator at any time through the control center and transmit the captured images back to the control center. At the same time, the operator can formulate an observation plan in the control center and automatically realize timed shooting. This operation does not require opening the incubator and the operator does not need to operate multiple times, which is convenient and fast.

[0023] (5) The present invention creates the low-energy laser-assisted cell culture and online monitoring device. The operator can set the corresponding program in the control center to plan laser irradiation and cell photography, so that the device can be executed for a long time and automatically, and collect images of the entire process of cell growth, reproduction, and differentiation, and draw a laser-cell growth curve, providing a convenient means for exploring the optimal laser irradiation conditions (including but not limited to: wavelength, energy density, duration, interval, frequency, etc.). BRIEF DESCRIPTION OF THE DRAWINGS

[0024] The accompanying drawings, which constitute part of the present invention, are intended to provide a further understanding of the present invention. The exemplary embodiments of the present invention and their descriptions are intended to explain the present invention and do not constitute an undue limitation of the present invention. In the accompanying drawings:

[0025] Figure 1 This is a schematic structural diagram of the laser-assisted cell culture device according to an embodiment of the present invention;

[0026] Figure 2 This is a structural diagram of the light source body according to an embodiment of the present invention;

[0027] Figure 3 A schematic diagram of the structure of the functional body according to an embodiment of the present invention;

[0028] Figure 4 This is a schematic structural diagram of the cell online monitoring device according to an embodiment of the present invention;

[0029] Figure 5 This is a schematic structural diagram of the low-energy laser-assisted cell culture and online monitoring device according to an embodiment of the present invention;

[0030] Figure 6 This is a simplified structural diagram of the low-energy laser-assisted cell culture and online monitoring device described in an embodiment of the present invention.

[0031] Description of reference numerals:

[0032] 1. Light source body; 2. Functional body; 3. Cell culture chamber; 4. Laser modulation module; 5. LED light source; 6. Fiber coupler; 7. Light source chassis; 8. Laser; 9. Fiber; 10. Functional chassis; 11. Point light source emitter; 12. Stage; 13. Plane mirror; 14. Collimating lens; 15. Objective lens; 16. Camera; 17. Electric translation stage; 18. Cell culture chamber; 19. Control center; 20. Data cable; 21. Hub. DETAILED DESCRIPTION

[0033] In order to make the purpose, technical solutions and advantages of the present invention more clearly understood, the present invention is further described in detail below with reference to the accompanying drawings and specific embodiments. It should be understood that the specific embodiments described herein are only used to explain the present invention and do not constitute a limitation of the present invention.

[0034] It should be noted that, in the absence of conflict, the embodiments of the present invention and the features in the embodiments can be combined with each other.

[0035] In the description of the present invention, it should be understood that the terms "center", "longitudinal", "lateral", "up", "down", "front", "back", "left", "right", "vertical", "horizontal", "top", "bottom", "inside", "outside" and the like indicate orientations or positional relationships based on the orientations or positional relationships shown in the accompanying drawings, and are only for the convenience of describing the present invention and simplifying the description, rather than indicating or implying that the device or element referred to must have a specific orientation, be constructed and operated in a specific orientation, and therefore cannot be understood as a limitation on the present invention. In addition, the terms "first", "second" and the like are only used for descriptive purposes and cannot be understood as indicating or implying relative importance or implicitly indicating the number of the indicated technical features. Thus, features defined as "first", "second" and the like may explicitly or implicitly include one or more of the features. In the description of the present invention, unless otherwise specified, "multiple" means two or more.

[0036] In the description of the present invention, it should be noted that, unless otherwise expressly specified or limited, the terms "installed," "connected," and "connected" should be understood in a broad sense. For example, they can refer to fixed connections, detachable connections, or integral connections; they can refer to mechanical connections or electrical connections; they can refer to direct connections or indirect connections through an intermediate medium; and they can refer to internal connections between two components. Those skilled in the art can understand the specific meanings of the above terms in the present invention based on specific circumstances.

[0037] The present invention will be described in detail below with reference to the accompanying drawings and in combination with embodiments.

[0038] like Figure 1 As shown, the laser-assisted cell culture device described in the embodiment of the present invention includes a light source body 1, a functional body 2, and a cell culture chamber 3. The light source body 1 is arranged outside the cell culture chamber 3 to prevent the high-power heating components in the light source body 1 from affecting the environment inside the cell culture chamber 3. The functional body 2 is arranged inside the cell culture chamber 3, and the cells to be cultured are placed in the functional body 2, so that the functional body 2 can irradiate and culture the cells to be cultured in the cell culture chamber 3 for a long time.

[0039] like Figure 2 As shown, the light source body 1 includes a laser modulation module 4, an LED light source 5, a fiber coupler 6, and a light source chassis 7. The laser modulation module 4, the LED light source 5, and the fiber coupler 6 are arranged in the light source chassis 7. The laser modulation module 4 emits no less than two beams of low-energy laser light to irradiate the cells to be cultured. The energy density of the low-energy laser light is (0.005~5) W / cm 2 , and the wavelength of each low-energy laser is different.

[0040] The laser modulation module 4 includes at least two lasers 8. In the embodiment of the present invention, the laser modulation module 4 includes two independent lasers 8, each of which generates lasers with wavelengths of 660 nm and 808 nm, respectively. Each laser 8 is equipped with a knob and a data interface, and an operator can manually rotate the knob to control the on / off state and / or energy density of the low-energy laser light of each laser 8.

[0041] LED light source 5 emits LED light to illuminate the cells to be cultured, providing good illumination for imaging the cells to be cultured when monitoring them, while avoiding the speckle effect caused by low-energy lasers. Fiber coupler 6 couples the LED light and all low-energy lasers into optical fiber 9. Optical fiber 9 passes through a pre-set hole in the side wall of the cell culture chamber 3 and connects to the functional body 2, allowing the optical fiber to transmit the coupled light beam into the functional body 2. Due to the small diameter of optical fiber 9, it can pass through the pre-set hole well without affecting the internal environment of the cell culture chamber 3. If necessary, a sealing ring can be added to the pre-set hole for additional sealing.

[0042] In the embodiment of the present invention, a hub 21 is further provided in the light source body 1. External devices can control the two lasers 8 and the LED light source 5 respectively through the data line 20 and the hub 21 to complete the adjustment of the output beams of the LED light source 5 and the laser 8.

[0043] like Figure 3 As shown, the functional body 2 includes a functional chassis 10 , and a point light source emitter 11 , a stage 12 , a plane reflector 13 and a collimating lens 14 arranged in the functional chassis 10 .

[0044] The surface of the functional chassis 10 is provided with a sealing device to protect the internal devices from the influence of the humid environment, such as corrosion. The point light source emitter 11 receives the coupled light beam transmitted through the optical fiber 9, and emits the coupled light beam in the form of a point light source. The specific process is as follows: one end of the optical fiber 9 couples the LED light and all low-energy lasers into the optical fiber 9 through the optical fiber coupler 6, and the other end of the optical fiber 9 is connected to the point light source emitter 11, and the point light source emitter 11 emits the coupled light beam in the form of a point light source. By controlling the light-on state of the LED light source 5 and each laser 8, the point light source emitter 11 can freely switch the output light between low-energy lasers or LED lights of different bands.

[0045] The diverging light beam is reflected by a plane reflector 13 and then into a collimating lens 14. The collimating lens 14 converts the reflected light into a parallel beam. This parallel beam illuminates the cells to be cultured, which are placed on the stage 12. The cells to be cultured are placed on the stage 12 in a cell culture cassette 18. In actual operation, the cell culture cassette 18 can be replaced with another vessel, such as a culture dish, that can hold the cells to be cultured, as needed.

[0046] In this embodiment of the present invention, to reduce the size of the illumination, the illumination light path of point light source emitter 11 is oriented horizontally. A plane reflector 13 is positioned at a 45-degree angle to the horizontal directly above collimating lens 14. Point light source emitter 11 is located at the equivalent focal point of collimating lens 14. The emitted light is reflected by plane reflector 13 and modulated into a parallel beam by collimating lens 14. Stage 12 is perpendicular to the propagation direction of the parallel beam.

[0047] The embodiment of the present invention also provides a cell online monitoring device, such as Figure 4 As shown, it includes an objective lens 15, a camera 16 and an electric translation stage 17. The light reflected by the cells to be cultured enters the camera 16 through the objective lens 15, so that the cells to be cultured are imaged on the target surface of the camera 16, completing the capture of the cell image. The camera 16 is placed on the electric translation stage 17, and the electric translation stage 17 changes the distance between the camera 16 and the cells to be cultured to complete the focusing of the camera 16. Furthermore, an attenuation plate and a quarter-wave plate are plated at the center of the back focal plane of the objective lens 15, and the attenuation plate and the quarter-wave plate match the parallel light beam. Since the outgoing light is reflected by the plane reflector 13 and then modulated into a parallel light beam by the collimating lens 14, the parallel light beam will naturally converge to the center of the back focal plane of the objective lens 15. Therefore, in an embodiment of the present invention, a circular attenuation plate is placed at the center of the back focal plane of the objective lens 15, which can also achieve interference between the parallel light beam and the scattered light of the cells to be cultured, produce phase contrast, and thus improve the imaging contrast of transparent cells.

[0048] The objective lens 15 used in this embodiment is a phase contrast objective lens with a magnification of 4 times, and the camera 16 is a Daheng board-level camera VEN-505-36U3M, which is placed inverted on the electric translation stage 17.

[0049] Based on the laser-assisted cell culture device and cell online monitoring device provided, the present invention also proposes a laser-assisted cell culture and online monitoring device, such as Figure 5 and Figure 6 As shown, it specifically includes a laser-assisted cell culture device, a cell online monitoring device and a control center 19.

[0050] Control center 19 controls laser modulation module 4, causing each laser 8 therein to emit low-energy laser light. It also controls LED light source 5 to emit LED light. Control center 19 is connected to camera 16, causing it to transmit captured cell images to control center 19. Control center 19 also controls motorized translation stage 17, causing it to adjust the distance between camera 16 and the cells to be cultured, thereby adjusting the focus of camera 16.

[0051] In this embodiment of the present invention, a control center 19 controls the two lasers 8 and the LED light source 5, respectively, via a data cable 20 and a hub 21. The camera 16, the motorized translation stage 17, and the control center 19 are connected via the data cable 20. The control center 19 is a host computer running Windows 10. It controls the laser modulation module 4, the LED light source 5, the camera 16, and the motorized translation stage 17, and stores cell images transmitted by the camera 16. The control center 19 operates the motorized translation stage 17, causing it to fine-tune the position of the camera 16 to achieve focus for imaging the cultured cells. The control center 19 also has a timing function and a program that sets the wavelength, energy density, duration, interval, and frequency of laser irradiation and imaging of cells. This program allows the control center to automatically and regularly control the laser modulation module 4, the LED light source 5, and the camera 16 to perform the specified tasks. The control center also has a data analysis function, analyzing cell images and, combined with current laser control data, plotting laser-cell growth curves.

[0052] When using the laser-assisted cell culture device and cell online monitoring device described in the embodiments of the present invention, the light source body 1 is placed outside the cell culture box 3, the functional body 2 is placed inside the cell culture box 3, and the optical fiber 9 is properly connected. The data cable 20 is properly connected to the control center 19, and then a cell culture box 18 or a culture dish or other device for holding cells to be cultured is placed on the stage 12. The operator can arbitrarily adjust the height of the electric translation stage 17 in the control center 19 to manually focus the camera 16, or can call the autofocus algorithm to achieve automatic focus and follow focus.

[0053] After completing the assembly of the device and adjusting the position of the components in the device, the operator can formulate an irradiation plan and a monitoring plan on the control center 19. When the planned task time arrives at Beijing time, the control center 19 will automatically control the device provided in the embodiment of the present invention to perform the low-energy laser irradiation function or the cell image shooting function. Among them, the irradiation plan and the monitoring plan provide flexible and rich parameters that are independent of each other, including the opening time, duration, energy density, irradiation frequency, cycle of each laser 8, and the shooting frequency, number of frames per shot, cycle, etc. of the camera 16. By customizing different irradiation plans for each laser 8, it is possible to achieve stable, long-term, repeated and unattended low-energy laser irradiation of cells in the box. The monitoring plan of the embodiment of the present invention will synchronously call the LED light source 5 and the camera 16, and the control center 19 will run the autofocus algorithm to achieve real-time focus locking of the camera 16, ensuring that the LED light source 5 is controlled to provide correct LED lighting and clear imaging while the camera 16 is exposed.

[0054] After the operator completes the configuration of the irradiation and monitoring plans in the control center 19, the formal culture process begins. During the formal culture process, the operator can monitor the real-time cell growth within the cell culture incubator 3 at any time on the control center 19. Historical images can also be viewed in a folder in the control center 19 to track the culture process. The culture and monitoring process does not require opening the cell culture incubator 3, thus avoiding environmental disturbances during cell growth, reducing the need for manual intervention, and simplifying the complex culture steps.

[0055] The planning period formulated in the embodiment of the present invention supports more than one month.

[0056] The laser-assisted cell culture device and the cell online monitoring device provided in the embodiments of the present invention can use the laser irradiation function or the cell monitoring function independently, and can be used as a separate low-energy laser irradiation instrument or a cell microscopic imaging instrument respectively.

[0057] It should be understood that the various forms of the processes shown above can be used to reorder, add, or delete steps. For example, the steps described in the present disclosure can be performed in parallel, sequentially, or in a different order, as long as the desired results of the technical solutions disclosed in the present disclosure can be achieved. This is not limited herein.

[0058] The above specific embodiments do not limit the scope of protection of the present invention. Those skilled in the art will appreciate that various modifications, combinations, sub-combinations, and substitutions may be made based on design requirements and other factors. Any modifications, equivalent substitutions, and improvements made within the spirit and principles of the present invention are intended to be included within the scope of protection of the present invention.

Claims

1. A laser-assisted cell culture device, characterized in that: The device comprises a light source body (1), a functional body (2) and a cell culture box (3), wherein the functional body (2) is arranged inside the cell culture box (3), and the light source body (1) is arranged outside the cell culture box (3); wherein cells to be cultured are placed in the functional body (2), and the light source body (1) transmits LED light for illuminating the cells to be cultured and low-energy laser light for irradiating the cells to be cultured to the functional body (2); The light source body (1) comprises a laser modulation module (4), an LED light source (5) and an optical fiber coupler (6); the laser modulation module (4) emits at least two beams of low-energy laser light for irradiating the cells to be cultured, and the wavelength of each beam of low-energy laser light is different; the LED light source (5) emits LED light for illuminating the cells to be cultured; the optical fiber coupler (6) couples the LED light and all the low-energy laser light into an optical fiber; and the optical fiber transmits the coupled light beams to the functional body (2).

2. The laser-assisted cell culture device according to claim 1, characterized in that: The laser modulation module (4) comprises no less than two lasers (8), each laser (8) emits low-energy laser light, and the wavelength of each low-energy laser light is different.

3. The laser-assisted cell culture device according to claim 1, characterized in that: The light source body (1) further comprises a light source chassis (7), and the laser modulation module (4), the LED light source (5) and the optical fiber coupler (6) are arranged in the light source chassis (7).

4. The laser-assisted cell culture device according to claim 1, characterized in that: The functional body (2) comprises a point light source emitter (11), a stage (12), a plane reflector (13) and a collimating lens (14); wherein the cells to be cultured are placed on the stage (12); the point light source emitter (11) receives a coupled light beam and irradiates the cells to be cultured with the coupled light beam in the form of a point light source; the light emitted by the point light source emitter (11) is reflected by the plane reflector (13) onto the collimating lens (14); the collimating lens (14) converts the reflected light into a parallel light beam; and the parallel light beam irradiates the cells to be cultured.

5. The laser-assisted cell culture device according to claim 4, characterized in that: The functional body (2) further comprises a functional chassis (10), wherein the point light source emitter (11), the stage (12), the plane reflector (13) and the collimating lens (14) are arranged in the functional chassis (10), and the surface of the functional chassis (10) is sealed.

6. The laser-assisted cell culture device according to claim 4, characterized in that: A reserved hole is provided on the side wall of the cell culture box (3), so that the optical fiber (9) extends into the cell culture box (3) through the reserved hole. The optical fiber coupler (6) couples the LED light and all low-energy lasers into the optical fiber (9). The other end of the optical fiber (9) is connected to the point light source emitter (11), and the point light source emitter (11) emits the coupled light beam in the form of a point light source.

7. A laser-assisted cell culture and online monitoring device, comprising the laser-assisted cell culture device according to any one of claims 1 to 6; characterized in that: It also includes a cell online monitoring device and a control center (19); wherein: The cell online monitoring device comprises an objective lens (15), a camera (16) and an electric translation stage (17); wherein light reflected by the cells to be cultured enters the camera (16) through the objective lens (15), so that the cells to be cultured are imaged on a target surface of the camera (16); the camera (16) is placed on the electric translation stage (17); the electric translation stage (17) changes the distance between the camera (16) and the cells to be cultured, thereby completing the focusing of the camera (16); The control center (19) controls the laser modulation module (4) so ​​that each laser (8) in the laser modulation module (4) emits the low-energy laser; the control center (19) controls the LED light source (5) to emit the LED light; the control center (19) is connected to the camera (16) so that the camera (16) transmits the captured cell image to the control center (19); The control center (19) controls the electric translation stage (17) so that the electric translation stage (17) changes the distance between the camera (16) and the cells to be cultured, thereby completing the focusing of the camera (16).

8. The laser-assisted cell culture and online monitoring device according to claim 7, characterized in that: An attenuation plate and a quarter-wave plate are plated at the center of the rear focal plane of the objective lens (15), so that the parallel light beam forms phase contrast interference after passing through the objective lens (15).

9. The laser-assisted cell culture and online monitoring device according to claim 7, characterized in that: The control center (19) has a timing function and is equipped with a setting program, wherein the setting program is used to set the wavelength, energy density, duration, interval and frequency of laser irradiation and cell photography, so that the control center (19) can automatically and regularly control the laser modulation module (4), the LED light source (5) and the camera (16) to perform the set tasks; the control center (19) has a data analysis function, analyzes the cell image, and draws a laser-cell growth curve in combination with the current laser control data.

Images