Multi-time-period sampler for bacteriophage
By designing an automated sampler for multiple time periods, and employing a motor-driven rack and pinion system and a cryogenic chamber, the problem of tedious multi-time period sampling in phage culture was solved, achieving efficient and accurate sample collection and preservation, and reducing the frequency of manual operation.
Patent Information
- Application Number
- CN202422979292.X
- Authority / Receiving Office
- CN · China
- Patent Type
- Utility models(China)
- Current Assignee / Owner
- Filing Date
- 2024-12-04
- Publication Date
- 2025-11-11
- Estimated Expiration
- 2034-12-04
AI Technical Summary
In current phage culture processes, sampling at multiple time points is cumbersome, requiring frequent manual opening of the culture device for sampling, which leads to a high risk of cross-contamination and low efficiency.
Design a multi-time-period automatic sampler that uses a forward and reverse motor to drive a gear and rack system to achieve timed and quantitative sampling. The sampling holes are isolated by a circular blocking plate and a baffle, and the sample activity is maintained by a cryogenic liquid box. A cleaning system is also provided to reduce manual operation.
It enables timed, quantitative, and cross-contamination-free sampling, improving sampling efficiency and accuracy, reducing the complexity of manual operations, and ensuring the activity of samples and the accuracy of experimental results.
Smart Images
Figure CN223535091U_ABST
Abstract
Description
Technical Field
[0001] This invention belongs to the field of phage culture technology, specifically relating to a multi-time period sampler for phages. Background Technology
[0002] Bacteriophages are ubiquitous organisms that exclusively use bacteria as hosts. They are a mass of genetic material encased in a protein coat, and most bacteriophages also have a "tail" to inject genetic material into the host.
[0003] The overuse and misuse of antibiotics in medicine and agriculture have promoted the spread of drug-resistant genes, exacerbating the global problem of bacterial resistance and leading to an increasing number of bacteria developing resistance to commonly used antibiotics, forming "superbugs." Infections caused by these drug-resistant bacteria not only increase the difficulty of patient treatment and medical costs but also pose a serious threat to public health. Therefore, there is an urgent need to find alternative antibacterial methods. Bacteriophage inhibition, due to its high specificity, ability to precisely target specific bacteria without affecting beneficial bacteria, and low side effects, has become a potential solution. Furthermore, bacteriophages can be used in combination with antibiotics to enhance efficacy, and because they are naturally occurring viruses, they possess good biocompatibility and low immune response, making them suitable for a variety of patients. At the same time, the preparation and formulation of bacteriophages are relatively simple, offering potential for personalized treatment. Therefore, to realize the clinical application of bacteriophages, we need to conduct bacteriophage culture work.
[0004] The basic steps for culturing bacteriophages typically include the following key steps: (1) Selecting host bacteria; Selecting host bacteria that correspond to the target bacteriophage. Ensure that the host bacteria are culturable and are the specific targets of the bacteriophage for infection; (2) Obtaining bacteriophages; Bacteriophages can be extracted from the natural environment (such as soil, water sources, etc.) or known bacteriophage strains can be used; (3) Isolating bacteriophages; Isolating bacteriophages by dilution or filtration, diluting the sample, and mixing it with the host bacteria to observe whether plaques are formed, thereby determining the presence of bacteriophages; (4) Culturing bacteriophages; Co-culturing bacteriophages with the host bacteria in a culture medium. Commonly used culture media include LB liquid medium or other culture media suitable for the host bacteria. The incubation time is generally several hours to several days. Then, the host bacteria are filtered through a 0.22um filter membrane to obtain the phage culture medium; (5) Quantification and identification: After gradient dilution, the concentration of phages is quantified by determining the plaque forming unit (PFU) method using the double-layer plate method, and molecular biological identification, such as PCR or sequencing, is performed to confirm their species and characteristics; (6) Storage of phages: The extracted phages are stored under suitable conditions, usually frozen at -80°C to maintain their activity.
[0005] Currently, the sampling method for bacteriophages involves: at the required sampling time, using a pipette to draw 1 ml of sample, filtering, and then detecting the titer. Furthermore, because it's necessary to monitor titer changes during phage culture and application, we need to take samples at fixed times and locations, repeatedly testing the growth status of the phage or its host bacteria during culture. Currently, phage sampling is usually done directly on the surface of the culture medium. For example, to test the phage's inhibitory effect on bacterial growth, we need to directly insert a sampler into the sample, sending the sample-coated swab tip for testing, or placing it in a constant temperature environment at 4°C for unified testing. To detect the impact of phages on bacteria at different time points, testing is typically required every 10-30 minutes, necessitating multiple samplings. Currently, recording the time and manually sampling at regular intervals is required. Sampling now necessitates opening the culture device and dipping the sample into the bacterial culture medium, a rather cumbersome process. Utility Model Content
[0006] The present invention aims to provide a multi-time period sampler for bacteriophages, solving the technical problem of cumbersome manual sampling operations for multiple time periods in existing bacteriophage culture.
[0007] To solve the above-mentioned technical problems, the present invention adopts the following technical solution:
[0008] A multi-time-period sampler for bacteriophages includes a base, a bottom frame and a rectangular slot on the upper surface of the base, the bottom frame is fixed on the base, the rectangular slot is located at the center of the bottom frame, and a cell culture container is placed in the rectangular slot.
[0009] The top of the bottom frame is equipped with a multi-time automatic sampling mechanism, which includes a middle box and three sets of rack and pinion sampling drive mechanisms. The middle box is a box structure with an open bottom. A sampling box is fixed inside the middle box. Three through sleeves are fixed on the top of the sampling box. Three sampling holes are opened on the bottom of the sampling box. The three sampling holes and the three through sleeves are arranged vertically and vertically. A circular blocking plate is installed at each sampling hole. The circular blocking plate is used to block the sampling hole. The circular blocking plate is hinged to the side wall of the sampling hole by a pin. A torsion spring is set on the pin. The torsion spring is used to reset the circular blocking plate.
[0010] Three sets of rack and pinion sampling drive mechanisms are arranged side by side on the sampling box. Each set of rack and pinion sampling drive mechanisms includes a forward and reverse motor, a limit switch and a sampling rod. The sampling rods of the three sets of rack and pinion sampling drive mechanisms are respectively inserted into three sleeves. The forward and reverse motor and the limit switch are installed on the top surface of the sampling box. Gears are installed on the main shaft of the forward and reverse motor. A rack is provided on the side of the sampling rod. The gear meshes with the rack. A sampling head is provided at the lower end of the sampling rod and a contact is provided at the upper end of the sampling rod. The contact is located directly above the limit switch.
[0011] The top of the middle box is equipped with a coolant box, which has a filling port; the base is also equipped with a controller and an atomizer module. The atomizer module is connected to an atomizer tube through a pipe. The top of the atomizer tube has an atomizing liquid port. The atomizer tube is connected to the side wall of the bottom frame through a flexible hose.
[0012] The controller is connected to the atomizer module and three forward and reverse motors. The atomization adjustment knob is located on the side wall of the base and is used to control the atomizer module.
[0013] Furthermore, an LED strip is provided on the upper surface of the base, which surrounds the outer perimeter of the base frame, and a lighting switch for controlling the LED strip is provided on the side wall of the base.
[0014] Furthermore, the inner wall of the sampling box is provided with a sponge layer, which absorbs physiological saline.
[0015] Furthermore, the bottom surface of the sampling box is fixed with two baffles, and the sampling hole in the middle is located between the two baffles.
[0016] Furthermore, a cleaning frame is provided between the middle box and the bottom frame, the cleaning frame is connected to a cleaning inlet pipe, and the bottom frame is connected to a drain pipe.
[0017] Compared with existing technologies, the advantages of this invention are as follows: This invention uses a forward and reverse reversible motor-driven gear and rack system to control the sampling rod's movement, enabling the timely and quantitative acquisition of samples from cell culture media; baffles are installed inside the sampling box to isolate different sampling holes, and a self-resetting circular plug is used to seal the sampling holes, effectively preventing cross-contamination between samples; the cryoprotectant box provides low-temperature preservation conditions for the collected samples, helping to maintain sample activity; the device also incorporates a design specifically for cleaning the internal space of the bottom frame, which not only facilitates daily cleaning and maintenance of the equipment but also further ensures the accuracy of experimental results; this invention reduces the frequency and complexity of manual operation through automation, while improving the accuracy and efficiency of sampling. Attached Figure Description
[0018] The accompanying drawings are provided to further illustrate the present invention and form part of the specification. They are used together with the embodiments of the present invention to explain the present invention, but do not constitute a limitation thereof. In the drawings:
[0019] Figure 1 A schematic diagram of a multi-time-period sampler for bacteriophages;
[0020] Figure 2 This is a schematic diagram of the cleaning frame and the bottom frame;
[0021] Figure 3 This is a schematic diagram showing the positions of the middle chamber and the sampling box;
[0022] Figure 4 This is a schematic diagram of a multi-time automatic sampling mechanism. Detailed Implementation
[0023] The technical solutions of the present utility model will be clearly and completely described below with reference to the accompanying drawings of the embodiments. Obviously, the described embodiments are only some embodiments of the present utility model, and not all embodiments. Based on the embodiments of the present utility model, all other embodiments obtained by those of ordinary skill in the art without creative effort are within the protection scope of the present utility model.
[0024] The present invention will be further described in detail below with reference to the embodiments.
[0025] like Figure 1-4 As shown, a specific embodiment of the multi-time-period sampler for bacteriophages provided by this utility model is as follows:
[0026] A multi-time sampling device for bacteriophages includes a base 1, a bottom frame 2 and a rectangular slot 3 on the upper surface of the base 1, the bottom frame 2 is fixed on the base 1, the rectangular slot 3 is located at the center of the bottom frame 2, and a cell culture container 4 is placed in the rectangular slot 3.
[0027] The top of the bottom frame 2 is equipped with a multi-time automatic sampling mechanism, which includes a middle box 5 and three sets of rack and pinion sampling drive mechanisms. The middle box 5 is a box structure with an open bottom. A sampling box 6 is fixed inside the middle box 5. Three through sleeves 7 are fixed on the top of the sampling box 6. Three sampling holes 8 are opened on the bottom of the sampling box 6. The three sampling holes 8 are arranged vertically and vertically with the three through sleeves 7. A circular blocking plate 9 is installed at each sampling hole 8. The circular blocking plate 9 is used to block the sampling hole 8. The circular blocking plate 9 is hinged to the side wall of the sampling hole 8 by a pin. A torsion spring is provided on the pin. The torsion spring is used to reset the circular blocking plate 9. The hinge method of the torsion spring and the pin is a conventional technology in the field.
[0028] Three sets of rack and pinion sampling drive mechanisms are arranged side by side on the sampling box 6. Each set of rack and pinion sampling drive mechanisms includes a forward and reverse motor 10, a limit switch 11, and a sampling rod 12. The sampling rods 12 of the three sets of rack and pinion sampling drive mechanisms are respectively inserted into three sleeves 7. The forward and reverse motor 10 and the limit switch 11 are installed on the top surface of the sampling box 6. A gear 13 is installed on the main shaft of the forward and reverse motor 10. A rack 14 is provided on the side of the sampling rod 12. The gear 13 meshes with the rack 14. A sampling head 15 is provided at the lower end of the sampling rod 12. A contact 16 is provided at the upper end of the sampling rod 12. The contact 16 is located directly above the limit switch 11.
[0029] A coolant box 17 is installed on the top of the middle box 5, and a liquid filling port 18 is provided on the coolant box 17; a controller and an atomizer module are also provided in the base 1. The atomizer module is connected to an atomizer tube 19 through a pipe. An atomizer liquid port 20 is provided on the top of the atomizer tube 19. The atomizer tube 19 is connected to the side wall of the bottom frame 2 through a flexible hose 29.
[0030] The controller is connected to the atomizer module and three forward and reverse motors 10 respectively. The side wall of the base 1 is equipped with an atomization adjustment knob 21, which is used to control the atomizer module.
[0031] The specific culture and sampling process of this utility model is as follows: Three forward and reverse motors 10 are controlled by a controller, which can be a microcontroller. The controller controls the start-up time of the forward and reverse motors 10 by inputting a specific programming program. Initially, the cell culture container 4 contains cell culture medium. Then, liquid containing bacteriophages is added to the nebulizer tube 19 through the nebulizer inlet 20. The nebulizer module is started, and the nebulizer-containing vapor is slowly and continuously introduced into the bottom frame 2 through nebulization. The working power of the nebulizer module can also be controlled by the nebulization adjustment knob 21 to adjust the nebulization speed. After 10 minutes, the controller starts the first forward and reverse motor 10. The forward and reverse motor 10, through the meshing of gear 13 and rack 14, drives the sampling rod 12 downwards until the contact 16 at the upper end of the sampling rod 12 contacts the limit switch 11. When the reverse motor 10 stops working, the sampling rod 12 moves downwards, pushes open the circular blocking plate 9, and inserts itself into the cell culture medium. The sampling head 15 at the end dips into the sample. After sampling, the controller controls the forward and reverse motor 10 to rotate in the opposite direction. The sampling rod 12 moves upwards from the sampling hole 8 back into the sampling box 6. The circular blocking plate 9 seals the sampling hole 8 again under the action of the torsion spring. To avoid cross-contamination during sampling, two baffles 22 can be set on the bottom surface of the sampling box 6. The middle sampling hole 8 is located between the two baffles 22, and the baffles 22 isolate the three sampling holes 8 from each other. After waiting for another 10 minutes, the controller starts the second forward and reverse motor 10 to perform the corresponding sampling. Finally, after waiting for another 10 minutes, the controller starts the third forward and reverse motor 10 to perform the corresponding sampling. By performing multiple samplings, samples from three time periods can be obtained.
[0032] Preferably, a sponge layer 23 can be provided on the inner wall of the sampling box 6. The sponge layer 23 absorbs physiological saline and can ensure a relatively humid space inside the sampling box 6.
[0033] In addition, the cryo-liquid box 17 is used to hold cryo-liquid, which can provide a relatively low temperature environment for the sampled sample to maintain its activity.
[0034] A light strip 24 is also provided on the upper surface of the base 1. The light strip 24 is arranged around the outer perimeter of the base frame 2. A lighting switch 25 for controlling the light strip 24 is provided on the side wall of the base 1.
[0035] To facilitate cleaning of the interior of the bottom frame 2 after sampling, a cleaning frame 26 can be installed between the middle box 5 and the bottom frame 2. The cleaning frame 26 is connected to a cleaning inlet pipe 27, and the bottom frame 2 is connected to a drain pipe 28. During cleaning, cleaning liquid is introduced through the cleaning inlet pipe 27 and injected into the bottom frame 2 and the cleaning frame 26 to soak and clean their inner walls. After cleaning, the waste liquid is discharged through the drain pipe 28.
[0036] It should be noted that, in this document, terms such as “comprising,” “including,” or any other variations thereof are intended to cover non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements includes not only those elements but also other elements not expressly listed, or elements inherent to such process, method, article, or apparatus.
[0037] Although embodiments of the present invention have been shown and described, it will be understood by those skilled in the art that various changes, modifications, substitutions and alterations can be made to these embodiments without departing from the principles and spirit of the present invention, the scope of which is defined by the appended claims and their equivalents.
Claims
1. A multi-time-period sampler for bacteriophages, characterized in that: Includes a base, the upper surface of which is provided with a bottom frame and a rectangular slot. The bottom frame is fixed to the base, and the rectangular slot is located at the center of the bottom frame. A cell culture container is placed in the rectangular slot. The top of the bottom frame is equipped with a multi-time automatic sampling mechanism, which includes a middle box and three sets of rack and pinion sampling drive mechanisms. The middle box is a box structure with an open bottom. A sampling box is fixed inside the middle box. Three through sleeves are fixed on the top of the sampling box. Three sampling holes are opened on the bottom of the sampling box. The three sampling holes and the three through sleeves are arranged vertically and vertically. A circular blocking plate is installed at each sampling hole. The circular blocking plate is used to block the sampling hole. The circular blocking plate is hinged to the side wall of the sampling hole by a pin. A torsion spring is set on the pin. The torsion spring is used to reset the circular blocking plate. Three sets of rack and pinion sampling drive mechanisms are arranged side by side on the sampling box. Each set of rack and pinion sampling drive mechanisms includes a forward and reverse motor, a limit switch and a sampling rod. The sampling rods of the three sets of rack and pinion sampling drive mechanisms are respectively inserted into three sleeves. The forward and reverse motor and the limit switch are installed on the top surface of the sampling box. Gears are installed on the main shaft of the forward and reverse motor. A rack is provided on the side of the sampling rod. The gear meshes with the rack. A sampling head is provided at the lower end of the sampling rod and a contact is provided at the upper end of the sampling rod. The contact is located directly above the limit switch. The top of the middle box is equipped with a coolant box, which has a filling port; the base is also equipped with a controller and an atomizer module. The atomizer module is connected to an atomizer tube through a pipe. The top of the atomizer tube has an atomizing liquid port. The atomizer tube is connected to the side wall of the bottom frame through a flexible hose. The controller is connected to the atomizer module and three forward and reverse motors. The atomization adjustment knob is located on the side wall of the base and is used to control the atomizer module.
2. The multi-time-period sampler for bacteriophages according to claim 1, characterized in that: An LED strip is also provided on the upper surface of the base, which surrounds the outer perimeter of the base frame. A lighting switch for controlling the LED strip is provided on the side wall of the base.
3. A multi-time-period sampler for bacteriophages according to claim 1, characterized in that: The inner wall of the sampling box is lined with a sponge layer, which absorbs physiological saline.
4. A multi-time-period sampler for bacteriophages according to claim 1, characterized in that: The bottom surface of the sampling box is fixed with two baffles, and the sampling hole in the middle is located between the two baffles.
5. A multi-time-period sampler for bacteriophages according to claim 1, characterized in that: A cleaning frame is installed between the middle box and the bottom frame. The cleaning frame is connected to a cleaning inlet pipe, and the bottom frame is connected to a drain pipe.