3D printing biological material activity in-situ detection device and detection method

Abstract

The invention relates to a 3D printing biological material activity in-situ detection device and a detection method. The detection device comprises a biological material 3D printer, a laser, an optical fiber, an optical fiber head, an adjustable reflector and a camera, wherein the biological material 3D printer comprises a main body shell, a moving assembly, a printing nozzle and a printing platform; the fiber connects the laser and the fiber head; a collimation coupler is arranged on the optical fiber head, and laser emitted by the optical fiber head penetrates through the biological material sprayed by the printing spray head and then is reflected by the adjustable reflecting mirror to enter the camera; the camera transmits shot images to the computer. Compared with the prior art, components such as the laser, the adjustable reflector and the camera are arranged on the biological material 3D printer, so that dynamic detection and in-situ detection on a biological material sprayed by the printing nozzle are realized; the adjustable reflector is used for adjusting the angle of the laser, the collimating coupler is used for converting the laser into planar light, the fixed reflector is used for changing the path of the laser, and the size of the whole device is saved.

Description

Technical Field

[0001] The present invention relates to the technical field of 3D printed biomaterials, and in particular to an in-situ detection device and method for the activity of 3D printed biomaterials. Background Art

[0002] Understanding the cell activity and number of viable cells in 3D printed biomaterials is a prerequisite for printing the required products with quality and quantity, and is also an important reference for subsequent cell culture.

[0003] It is difficult to capture the cells contained in the working and flowing 3D bioprinting materials. The main difficulty lies in its real-time and accuracy. The methods currently used are mostly observation through traditional microscopes after staining. These two methods are more often used to monitor cell activity during cell culture. They have major defects in practical applications, specifically: (1) The distribution of cells is three-dimensional. Through traditional microscopes, only the distribution of cells in a certain plane can be seen, so the information of all cells cannot be obtained; (2) Traditional microscopes cannot obtain the activity of cells printed at a certain moment in real time. They can only be monitored during culture, and it is difficult to accurately obtain the cell information at that moment; (3) Cells are transparent. Traditional microscopes cannot observe unstained cells, but staining will cause certain damage to cells, making it difficult to observe cells when they are healthy and alive.

[0004] After searching, the application publication number CN113985593A discloses a portable coaxial digital holographic microscope and imaging method based on 3D printing technology, which specifically discloses: the device includes a base, a support plate connected to the upper end of the base, a light source loading slot connected to the upper end of the support plate, a light source connected inside the light source loading slot, an objective lens connected to the upper end of the base, a reflector loading platform, a reflector, and an image sensor connected inside, wherein the signal output end of the image sensor is connected to a computer. However, the prior art only discloses the structure of the microscope, and does not disclose how to monitor the activity of cells during 3D printing.

[0005] In summary, how to provide a device that can monitor cell activity in 3D printed biomaterials in real time is a technical problem that needs to be solved. Summary of the invention

[0006] The purpose of the present invention is to provide a 3D printed biomaterial activity in-situ detection device and method in order to overcome the defect of the above-mentioned prior art that it is difficult to obtain biomaterial activity information in real time.

[0007] The purpose of the present invention can be achieved by the following technical solutions:

[0008] According to one aspect of the present invention, there is provided a 3D printed biomaterial activity in-situ detection device, which is connected to a computer. The detection device comprises a biomaterial 3D printer, which comprises a main body shell and a moving component, a print nozzle and a printing platform installed in the main body shell, wherein the print nozzle and the printing platform are installed on the moving component, and the printing platform is located on the side where the print nozzle ejects the biomaterial and is perpendicular to the print nozzle. The detection device further comprises a laser, an optical fiber, an optical fiber head, an adjustable reflector and a camera installed in the main body shell; the two ends of the optical fiber are respectively connected to the laser and the optical fiber head; a collimating coupler is provided on the optical fiber head, and the laser emitted by the optical fiber head passes through the biomaterial ejected by the print nozzle, is reflected by the adjustable reflector, and enters the camera perpendicular to the camera lens; the camera transmits the captured image to the computer.

[0009] As a preferred technical solution, the detection device further includes a lens, which is mounted on one side of the camera lens, and the laser passes through the lens and enters the camera lens.

[0010] As a preferred technical solution, the detection device also includes a second bracket, and the adjustable reflector and the camera are installed on two opposite sides of the main shell through the second bracket; the connection direction of the adjustable reflector and the camera is perpendicular to the print head.

[0011] As a preferred technical solution, screw rods are installed on two opposite side surfaces in the main body shell, and the second bracket is installed on the screw rod and cooperates with the screw rod.

[0012] As a preferred technical solution, the camera is a charge coupled device camera.

[0013] As a preferred technical solution, the moving component includes a translation component and a lifting component. The print head is installed on the translation component and moves in the X and Y directions with the translation component; the printing platform is installed on the lifting component and moves in the Z direction with the lifting component.

[0014] As a preferred technical solution, the main shell is provided with threading holes, and the power lines and communication lines of the laser and camera pass through the threading holes and out of the main shell to be connected to the power supply and the computer respectively.

[0015] As a preferred technical solution, a transparent observation window is provided on the main body shell.

[0016] According to another aspect of the present invention, a detection method of a 3D printed biomaterial activity in-situ detection device is provided, which specifically comprises the following steps:

[0017] Step S101, the printing nozzle ejects the biological material and prints it vertically on the printing platform;

[0018] Step S102, the laser passes through the biological material and reaches the adjustable reflector, and part of the laser is scattered when passing through the biological material;

[0019] Step S103, the adjustable reflector reflects the laser to the camera, and the partially scattered laser interferes with the unscattered laser to form a holographic image;

[0020] Step S104, the camera captures the holographic image and transmits it to the computer;

[0021] Step S105: The computer obtains a particle dynamic image after model reconstruction.

[0022] Compared with the prior art, the present invention has the following beneficial effects:

[0023] 1) The present invention realizes dynamic detection and in-situ detection of biomaterials ejected from a print head by arranging components such as a laser, an adjustable reflector, and a camera on a biomaterial 3D printer, thereby obtaining activity data of the biomaterial; the adjustable reflector is used to adjust the angle so that the laser enters the camera vertically, and the collimating coupler is used to convert the laser into planar light;

[0024] 2) The present invention sets a fixed reflector. When the laser emitted by the optical fiber head is parallel to the printing nozzle, the fixed reflector can reflect the laser to the biological printing material. By changing the laser path through the fixed reflector, the volume of the entire device can be saved;

[0025] 3) The lens of the present invention can magnify the holographic image to make the image clearer; the adjustable reflector and the camera are installed on opposite sides to ensure that the light path is not blocked; the screw rod can be set to adjust the position of the adjustable reflector and the camera; the charge coupled device camera is used, which has higher sensitivity to light;

[0026] 4) The present invention is provided with a threading hole, through which the power line and the communication line in the device can be conveniently passed out; a transparent observation window is provided to observe the situation inside the biomaterial 3D printer at any time. BRIEF DESCRIPTION OF THE DRAWINGS

[0027] Figure 1 This is a schematic diagram of the overall structure of a 3D printed biomaterial activity in-situ detection device of the present invention;

[0028] Figure 2 This is a schematic diagram of the structure of the present invention without the main housing and the moving components;

[0029] Figure 3 Experimental diagram of collecting gelatin for the present invention;

[0030] Figure 4 This is an experimental diagram of the present invention using the same lens at different collection distances;

[0031] Figure 5 This is an experimental diagram of the particle situation at a certain depth of focus of the present invention;

[0032] Figure 6 This is a flow chart of a method for in-situ detection of activity of 3D printed biomaterials according to the present invention;

[0033] The numbers in the figure show:

[0034] 11. Main shell, 121. Translation assembly, 122. Lifting assembly, 13. Print nozzle, 14. Printing platform, 15. First bracket, 16. Second bracket, 17. Screw, 2. Laser, 3. Optical fiber, 4. Optical fiber head, 5. Adjustable reflector, 6. Camera, 7. Fixed reflector, 8. Lens. DETAILED DESCRIPTION

[0035] The following will be combined with the drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work should fall within the scope of protection of the present invention.

[0036] Example 1

[0037] like Figure 1 As shown, the present invention provides a 3D printed biomaterial activity in situ detection device, including a biomaterial 3D printer, a laser 2, an optical fiber 3, an optical fiber head 4, an adjustable reflector 5, a fixed reflector 7, a lens 8, a camera 6, a first bracket 15 and a second bracket 16.

[0038] The biomaterial 3D printer is a three-axis printer that can provide movement in three directions, including a main housing 11, a moving component, a print head 13, and a printing platform 14. The moving component, the print head 13, and the printing platform 14 are all installed in the main housing 11. The moving component includes a translation component 121 and a lifting component 122. The print head 13 is installed on the translation component 121 and moves with the translation component 121 in the X direction and the Y direction; the printing platform 14 is installed on the lifting component 122 and moves with the lifting component 122 in the Z direction. The print head 13 sprays out the biomaterial for 3D printing, and the printing platform 14 is located directly below the print head 13 and is perpendicular to the print head 13.

[0039] The lifting assembly 122 includes a first screw rod 17, a second screw rod 17, a third screw rod 17 and a first fixing member, and is connected to the main housing 11 through the first fixing member; the translation assembly 121 includes a first slider, a first chain, a first sprocket, a first sprocket connecting member, a second slider, a second chain, a second sprocket, a second sprocket connecting member, a third sprocket connecting member, a third slider, a fourth slider, a fifth slider and a sixth slider for providing Y-direction movement; the first slider is connected to the first chain, and the first chain is connected to the main housing 11 through the first sprocket, the first sprocket connecting member and the second slider; the second chain is connected to the second sprocket connecting member and the third sprocket connecting member through the second sprocket; the third slider, the fourth slider, the fifth slider and the sixth slider together provide Y-direction movement; the translation assembly 121 also includes a fourth screw rod 17 and a second fixing member for providing X-direction movement, and the fourth screw rod 17 is connected to the housing through the second fixing member and the first slider, and is driven by a motor.

[0040] like Figure 2 As shown, the laser 2 is installed in the main housing 11 and connected to one end of the optical fiber 3, and the other end of the optical fiber 3 is connected to the optical fiber head 4. The laser 2 can also be installed outside the main housing 11.

[0041] The optical fiber head 4 is provided with a collimating coupler to convert the emitted laser into planar coherent light; the optical fiber head 4 is installed on one side of the print nozzle 13, and the direction of the light emitted by the optical fiber head 4 is the same as the direction in which the print nozzle 13 ejects the biological material. The print nozzles 13 are fixed to the moving assembly through the first bracket 15.

[0042] The fixed reflector 7 is mounted on the first bracket 15 and is located below the optical fiber head 4 . The laser emitted by the optical fiber head 4 is reflected by the fixed reflector 7 and then passes through the biological material ejected by the printing nozzle 13 .

[0043] The adjustable reflector 5 and the camera 6 are mounted on two opposite sides of the main housing 11 through the second bracket 16. The two sides are provided with screws 17. The second bracket 16 is mounted on the screws 17 and cooperates with the screws 17. The cells are cultured in gelatin and ejected vertically from the nozzle of the printer, so the connecting direction of the adjustable reflector 5 and the camera 6 is perpendicular to the print head 13. The laser passes through the biological material ejected from the print head 13 and reaches the adjustable reflector 5. The adjustable reflector 5 can fine-tune the angle to ensure that the nozzle can accurately receive the image at any position.

[0044] The lens 8 is installed in front of the camera 6 to magnify the image, including a beam expander lens 8 and a magnifying lens 8.

[0045] The camera 6 is a charge coupled device camera 6 (CCD for short). A charge coupled device is a semiconductor device that can convert light into electric charge, store and transfer the electric charge. The laser reflected by the adjustable reflector 5 enters the camera 6 perpendicular to the lens of the camera 6. The camera 6 is connected to a computer, and the image collected by the camera 6 is transmitted to the computer for image reconstruction.

[0046] The surface of the main housing 11 is also provided with a transparent observation window and a threading hole, through which the power lines and communication lines of the laser 2 and the camera 6 pass through the threading holes and are connected to the power supply and the computer respectively. The transparent observation window can observe the 3D printing process and the coaxial digital holographic recording process.

[0047] Example 2

[0048] like Figure 6 As shown, the present invention provides a method for in situ detection of activity of 3D printed biomaterials. Gelatin material containing cells is ejected from a printing nozzle. Plane coherent light is passed through the gelatin to irradiate the cells contained therein. Part of the plane coherent light is scattered by the gelatin. The scattered light interferes with the unscattered direct light to form a hologram. After being magnified by a lens 8, the holographic image is collected by a CCD camera 6, and the image is input into a computer to reconstruct a clear image of the material.

[0049] In order to verify the feasibility of the in-situ detection method of the present invention, the following experiment was conducted, and a holographic test was performed on a clean bench. The holographic image was reconstructed to obtain an in-focus image of the particles in the gelatin. The relevant experimental parameters are as follows:

[0050] Gelatin specifications: cylinder with a diameter of 0.6 mm;

[0051] Laser wavelength: 632nm;

[0052] Spatial filter 2-2 parameters: 40x microscope objective, 25um pinhole;

[0053] Specifications of beam expander lens 82-4: diameter φ = 50.8 mm, focal length f = 200 mm;

[0054] Magnifying lens 8: magnification 4 times;

[0055] Camera 6 pixel size: 2.2um×2.2um.

[0056] like Figure 3 Shown is a holographic image of the distribution of particles in gelatin, in which the particles contained can be clearly seen.

[0057] like Figure 4 As shown in the figure, by selecting different collection distances, it can be found that the particles contained in them cannot be seen

[0058] like Figure 5 As shown, Figure 3 The collected images are reconstructed at a reconstruction distance of 1 mm to obtain the particles focused on the plane. Similarly, by selecting multiple planes and reconstructing them separately, the particle distribution on all planes can be obtained. By combining the particle distributions obtained from all planes, all the particles contained in the gelatin at that moment can be obtained.

[0059] It can be seen from the holographic measurement results that the present invention works normally, the parameter settings are reasonable, and the expected experimental effect is achieved. This shows that the application of the present invention in the measurement of 3D printing of biological materials is successful. During the experiment, the magnification of the imaging system and the acquisition distance can be changed to capture the clear dynamics of particles in different particle size ranges while ensuring the measurement accuracy.

[0060] The above is only a specific embodiment of the present invention, but the protection scope of the present invention is not limited thereto. Any technician familiar with the technical field can easily think of various equivalent modifications or replacements within the technical scope disclosed by the present invention, and these modifications or replacements should be included in the protection scope of the present invention. Therefore, the protection scope of the present invention shall be based on the protection scope of the claims.

Claims

1. A 3D printed biomaterial activity in-situ detection device, connected to a computer, the detection device comprising a biomaterial 3D printer, the biomaterial 3D printer comprising a main body shell (11) and a moving component installed in the main body shell (11), a print head (13) and a printing platform (14), the print head (13) and the printing platform (14) are installed on the moving component, the printing platform (14) is located on the side of the print head (13) ejecting the biomaterial and is perpendicular to the print head (13); characterized in that: The detection device further comprises a laser (2), an optical fiber (3), an optical fiber head (4), an adjustable reflector (5) and a camera (6) installed in a main body housing (11); the two ends of the optical fiber (3) are respectively connected to the laser (2) and the optical fiber head (4); a collimating coupler is provided on the optical fiber head (4); the laser light emitted by the optical fiber head (4) passes through the biological material ejected by the printing nozzle (13), is reflected by the adjustable reflector (5), and enters the camera (6) perpendicular to the lens of the camera (6); the camera (6) transmits the captured image to a computer.

2. A 3D printed biomaterial activity in-situ detection device according to claim 1, characterized in that: The detection device further comprises a first bracket (15) and a fixed reflector (7), wherein the fixed reflector (7) and the print head (13) are fixed to the moving assembly via the first bracket (15); the optical fiber head (4) is mounted on one side of the print head (13) and emits laser light parallel to the print head (13); the laser light emitted by the optical fiber head (4) is reflected by the fixed reflector (7) and then passes through the biological material ejected by the print head (13).

3. The in-situ detection device for 3D printed biomaterial activity according to claim 1, characterized in that: The detection device also includes a lens (8), which is mounted on one side of the lens of the camera (6), and the laser passes through the lens (8) and enters the lens of the camera (6).

4. The in-situ detection device for 3D printed biomaterial activity according to claim 1, characterized in that: The detection device also includes a second bracket (16), and the adjustable reflector (5) and the camera (6) are mounted on two opposite side surfaces in the main housing (11) via the second bracket (16); the connecting direction of the adjustable reflector (5) and the camera (6) is perpendicular to the print head (13).

5. The in-situ detection device for 3D printed biomaterial activity according to claim 4, characterized in that: A screw rod (17) is installed on two opposite side surfaces in the main body shell (11), and the second bracket (16) is installed on the screw rod (17) and cooperates with the screw rod (17).

6. The in-situ detection device for 3D printed biomaterial activity according to claim 1, characterized in that: The camera (6) is a charge coupled device camera (6).

7. The in-situ detection device for 3D printed biomaterial activity according to claim 1, characterized in that: The moving assembly comprises a translation assembly (121) and a lifting assembly (122); the printing head (13) is mounted on the translation assembly (121) and moves in the X direction and the Y direction along with the translation assembly (121); the printing platform (14) is mounted on the lifting assembly (122) and moves in the Z direction along with the lifting assembly (122).

8. The in-situ detection device for 3D printed biomaterial activity according to claim 1, characterized in that: The main housing (11) is provided with a threading hole, and the power line and communication line of the laser (2) and the camera (6) pass through the threading hole and out of the main housing (11) to be connected to the power supply and the computer respectively.

9. The in-situ detection device for 3D printed biomaterial activity according to claim 1, characterized in that: The main body shell (11) is provided with a transparent observation window.

10. A detection method using the 3D printed biomaterial activity in-situ detection device according to claim 1, characterized in that: The specific steps include: Step S101, the printing nozzle (13) ejects the biological material and prints it vertically on the printing platform (14); Step S102, the laser passes through the biological material and reaches the adjustable reflector (5), and part of the laser is scattered when passing through the biological material; Step S103, the adjustable reflector (5) reflects the laser light to the camera (6), and the partially scattered laser light interferes with the unscattered laser light to form a holographic image; Step S104, the camera (6) collects the holographic image and transmits it to the computer; Step S105: The computer obtains a particle dynamic image after model reconstruction.

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