CN215103312U

CN215103312U - Material culture device and system suitable for researching biological rhythm

Info

Publication number: CN215103312U
Application number: CN202022572843.2U
Authority: CN
Inventors: 侯宏伟; 钱庆; 赵旭耀; 皇甫翔宇
Original assignee: Qingpingwan Wuhan Biotechnology Co ltd
Current assignee: Qingpingwan Wuhan Biotechnology Co ltd
Priority date: 2020-11-09
Filing date: 2020-11-09
Publication date: 2021-12-10
Anticipated expiration: 2030-11-09

Abstract

The utility model belongs to the technical field of time biology, specifically provide a be suitable for material culture apparatus and system of research biorhythm, including the sealed chamber, the bottom of sealed chamber is equipped with the rotatable objective table that can place a plurality of culture dishes, and the culture dish is used for cultivateing biological individual sample or cell/tissue sample, and the top of sealed chamber is equipped with the entrance port that is used for illumination and vegetation's light, exciting light entrance port and imaging lens, and the objective table is located the illumination light entrance port, exciting light entrance port and imaging lens's effect scope, and the side is equipped with into wind gap fan and air outlet fan respectively about the sealed chamber. The illumination control module is used for driving and adjusting illumination intensity and illumination time in the incubator. The fluorescent signal emitted by the sample is obtained by imaging to capture a change indicative of the biological rhythm. The device has simple structure and high automation degree.

Description

Material culture device and system suitable for researching biological rhythm

Technical Field

The utility model belongs to the technical field of time biology, concretely relates to be suitable for material culture apparatus and system of studying biological rhythm.

Background

Temporal biology is a hot spot in current physiological research, and there is a close link between circadian and behavioral changes of biorhythmicity and intrinsic gene expression of biological clocks. The data provided by the biorhythm recording instrument is the basis of the research of time biology, and provides powerful support for people to understand biorhythm. However, the current automatic recording instruments mostly focus on the physiological indexes, behavior changes and the like of animals, and a corresponding real-time monitoring instrument for the rhythmic expression of molecular biological clock/clock-controlled genes is lacked. Therefore, detecting and recording the expression of its biological clock/clock-controlled genes from the molecular level for living material is a central issue in current time biology.

The current long-range recording systems for animal biological clock research are more, mainly studied from individual-level physiological or activity rhythms, and as early as the seventies of the last century, biologists have begun to use the Drosophila's activity rhythm recording system for screening of individuals with biological clock genes mutant Drosophila, and successfully cloned into the Drosophila's biological clock genes Per (PERIOD) and Tim (TIMESS) (Konopka and Benzer, 1971; Sehgal et al, 1994; Sehgal et al, 1995). Automated long-range recording systems also play an important role in the biological clock research of mammals. The Takahashi laboratory analyzed the rhythm of autonomic activity of the mice by recording autonomic activity through a mouse wheel, and found mice with a mutation in the biological Clock gene, and finally cloned into the Clock gene (Gekakiset al, 1998). In addition, there is a body temperature recording system embedded in the abdominal cavity of a mouse for recording the body temperature rhythm of the mouse. These automated recording time accuracies are also on the order of minutes, with time spans of up to weeks or even months (Mei et al, 2018). But there are fewer recording systems for molecular level in animals (camile, andre et al 2013). In plants, although fluorescent reporter systems were introduced earlier for biorhythm studies, only individual laboratories have self-made simple automated long-range recording systems (Tomoaki et al, 2013), which are single in function and cannot fully meet the needs of circadian rhythm studies. With the deepening of the research of the biorhythm, more branches appear in the research field of the time biology, the internal molecular mechanism of the biorhythm is the most central theoretical basis of the research, and equipment for detecting the circadian rhythm from the molecular level, which has higher system integration, more complete functions and stronger universality, is required to be used.

Disclosure of Invention

The to-be-solved technical problem of the utility model is to provide a simple structure, with low costs are suitable for material culture apparatus and system of research biorhythm for solve above-mentioned technical problem.

For this, the utility model provides a be suitable for material culture apparatus of research biorhythm, including the sealed chamber, the bottom of sealed chamber is equipped with the rotatable objective table that can place a plurality of culture dishes, and the culture dish is used for cultivateing biological individual sample or cell/tissue sample, the top of sealed chamber is equipped with entrance, exciting light entrance and the imaging lens that are used for illumination and vegetation's light, the objective table is located in the effect scope of illumination light entrance, exciting light entrance and imaging lens, lay the culture dish on the objective table and be used for bearing and monitor the sample, the side is equipped with into wind gap fan and air outlet fan respectively about the sealed chamber.

Preferably, the top of the closed chamber is also provided with an ultraviolet lamp sterilizer, and the exciting light entrance port, the entrance port for the light used for illumination and plant growth and the ultraviolet lamp sterilizer are sequentially arranged from the middle to the side of the closed chamber.

Preferably, the outsides of the air inlet fan and the air outlet fan are respectively provided with a curved air duct for avoiding light.

Preferably, the illumination light entrance ports are symmetrically distributed along a center line of the closed chamber.

Preferably, the excitation light incident ports are symmetrically distributed along a center line of the closed chamber.

Preferably, a heater is further arranged in the closed chamber.

Preferably, a xenon lamp light source is arranged at the exciting light incidence opening.

Preferably, the culture dish includes a plurality ofly, the bottom of airtight chamber is equipped with rotatable objective table, each culture dish evenly arranges in rotatable objective table.

The utility model also provides a material culture system who is suitable for research biorhythm, including the signal processing module, still include as former any the material culture apparatus who is suitable for research biorhythm, the signal processing module is used for handling material culture apparatus's illumination light signal, exciting light signal and imaging lens signal.

The utility model has the advantages that: the utility model provides a material culture apparatus and system suitable for research biorhythm, including the sealed chamber, the bottom of sealed chamber is equipped with the rotatable objective table that can place a plurality of culture dishes, and the culture dish is used for cultivateing biological individual sample or cell tissue sample, and the top of sealed chamber is equipped with the entrance port that is used for illumination and vegetation's light, exciting light entrance port and imaging lens, and rotatable objective table is located the illumination light entrance port, exciting light entrance port and imaging lens's effect scope, and the side is equipped with into wind gap fan and air outlet fan respectively about the sealed chamber. The petri dish bears the monitored sample, keeps the environment sealed and provides favorable conditions for sample growth and imaging. The illumination control module is used for driving and adjusting illumination intensity and illumination time in the incubator. The temperature control module is used for controlling the environmental temperature in the incubator, the environmental temperature is increased and reduced through the temperature control device, and meanwhile, the light-resistant curved air duct is designed, so that the darkroom condition during fluorescence imaging is ensured. The sterilization module adopts the mode of ultraviolet irradiation to carry out the sterilization of whole environment, based on preceding degree tunable ultraviolet lamp and driver design, provides aseptic good living environment for the sample that is surveyed.

The present invention will be described in further detail with reference to the accompanying drawings.

Drawings

FIG. 1 is a schematic diagram of the structural principle of the material culturing device and system suitable for studying biological rhythm of the present invention;

FIG. 2 is a perspective view of the material culturing apparatus and system for studying biological rhythm of the present invention;

FIG. 3 is a schematic diagram of a fluorescence imaging module of the material culture apparatus and system of the present invention adapted for studying biological rhythms;

FIG. 4 is a schematic diagram of the signal processing module of the material culturing device and system for studying biological rhythm of the present invention.

Description of reference numerals: ultraviolet lamp sterilizer 1, illumination light incident port 2, exciting light incident port 3, imaging lens 4, curved wind channel 5, income wind gap fan 6, air outlet fan 7, culture dish 8, sealed chamber 9, xenon lamp light source 10, exciting light optical filter module 11, emission light optical filter module 12, SCMOS13, sample 14.

Detailed Description

The technical solutions in the embodiments of the present invention will be described clearly and completely with reference to the accompanying drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only some embodiments of the present invention, not all embodiments. Based on the embodiments in the present invention, all other embodiments obtained by a person skilled in the art without creative efforts belong to the protection scope of the present invention.

In the description of the present invention, it is to be understood that the terms "center", "upper", "lower", "front", "rear", "left", "right", "vertical", "horizontal", "top", "bottom", "inner", "outer", and the like indicate orientations or positional relationships based on those shown in the drawings, and are merely for convenience of description and simplicity of description, and do not indicate or imply that the device or element referred to must have a particular orientation, be constructed and operated in a particular orientation, and therefore, are not to be construed as limiting the present invention.

The terms "first", "second" and "first" are used for descriptive purposes only and are not to be construed as indicating or implying relative importance or implicitly indicating the number of technical features indicated. Thus, a feature defined as "first" or "second" may explicitly or implicitly include one or more of that feature; in the description of the present invention, "a plurality" means two or more unless otherwise specified.

As shown in fig. 1 and 2, the embodiment of the utility model provides a be suitable for material culture apparatus of studying biological rhythm, including sealed chamber 9, sealed chamber 9's bottom is equipped with the rotatable objective table that can place a plurality of culture dishes 8, and culture dish 8 is used for cultivateing biological individual sample 14 or cell/tissue sample 14, sealed chamber 9's top is equipped with entrance port, exciting light entrance port 3 and the imaging lens 4 that are used for illumination and vegetation's light, the objective table is located in the effect scope of illuminating light entrance port 2, exciting light entrance port 3 and imaging lens 4, lay culture dish 8 on the objective table and be used for bearing and monitor sample 14, side is equipped with into wind gap fan 6 and air outlet fan 7 respectively about sealed chamber 9.

The circadian rhythm monitoring system based on fluorescence imaging comprises a sample 14 culture module, a fluorescence imaging module and a signal processing module 3. The sample 14 culture module is composed of a sterilization module, an illumination control module, a temperature control module and a sample 14 culture box. The fluorescence imaging module consists of a visible light illumination module, a xenon lamp light source 10, an excitation light filter module 11, an emission light filter module 12 and SCMOS 13. The signal processing module is a computer system comprising computer hardware and software, wherein the software comprises image processing software and system control software. The image processing software is responsible for the fusion and analysis of the fluorescence image, and the system control software is used for the functions of human-computer interaction, imaging control, environmental control and the like. The sample 14 culture module is used for culturing a biological individual sample 14 or a cell/tissue sample 14 and is used as a carrying device for detecting the sample 14 by the system. The illumination and temperature are the most important time-service factors of plant circadian rhythm system, and have large correlation with the period, amplitude and dynamic balance of related genes of circadian rhythm. The sample 14 culture module can dynamically adjust the parameters, modulate the holographic growth and observation environment, and ensure the comprehensiveness and continuity of the biological rhythm scientific research.

Preferably, the top of the closed chamber 9 is further provided with an ultraviolet lamp sterilizer 1, and the exciting light entrance port 3, the entrance port for the light used for illumination and plant growth and the ultraviolet lamp sterilizer 1 are sequentially arranged from the middle to the side of the closed chamber 9. The sample 14 culture module has a sterilization function, sterilizes the internal environment of the system by adopting an ultraviolet sterilization mode, and ensures the growth environment of the detected sample 14 based on the design of a front-degree tunable ultraviolet lamp and a driver.

In a preferred scheme, the outsides of the air inlet fan 6 and the air outlet fan 7 are respectively provided with a curved air duct 5 for avoiding light. Due to the sensitivity of biorhythm to light and temperature, the culture module is required to have better sealing performance and isolate the influence of factors such as light, temperature and the like of the external environment on monitoring activities. The temperature control module is used for controlling the environmental temperature in the incubator, the environmental temperature is increased and reduced through the temperature control device, and the dark room condition during fluorescence imaging is ensured by designing the light-proof curved air duct 5.

Preferably, the illumination light inlets 2 are symmetrically distributed along a center line of the closed chamber 9. The excitation light incident ports 3 are symmetrically distributed along the center line of the closed chamber 9. The plurality of illumination light incident ports 2 or the plurality of excitation light incident ports 3 can better meet different illumination ranges and intensity requirements. The symmetrical distribution can also ensure the consistent illumination intensity of the culture dishes 8 in the closed chamber 9.

Preferably, a heater is further disposed in the sealed chamber 9. The temperature control module is used for controlling the ambient temperature in the incubator and increasing and reducing the ambient temperature through the temperature control device.

The embodiment of the utility model provides a material culture system who is suitable for research biorhythm still provides, including signal processing module, still include as former any the material culture apparatus who is suitable for research biorhythm, signal processing module is used for handling material culture apparatus's illumination light signal, exciting light signal and imaging lens 4 signal. The structure and the working principle are the same as those of the material culture device, and are not described in detail herein.

The following description of the working principle of a specific material culture device and system suitable for studying biological rhythms is provided: the material culture device and system suitable for researching biological rhythm comprises a sample 14 culture module, a fluorescence imaging module and a signal processing module 3. The sample 14 culture module is composed of a sterilization module, an illumination control module, a temperature control module and a sample 14 culture box. The fluorescence imaging module consists of a visible light illumination module, a xenon lamp light source 10, an excitation light filter module 11, an emission light filter module 12 and SCMOS 13. The signal processing module is a computer system comprising computer hardware and software, wherein the software comprises image processing software and system control software. The image processing software is responsible for the fusion and analysis of the fluorescence image, and the system control software is used for the functions of human-computer interaction, imaging control, environmental control and the like.

The sample 14 culture module is used for culturing a biological individual sample 14 or a cell/tissue sample 14 and is used as a carrying device for detecting the sample 14 by the system. The illumination and temperature are the most important time-service factors of plant circadian rhythm system, and have large correlation with the period, amplitude and dynamic balance of related genes of circadian rhythm. The sample 14 culture module can dynamically adjust the parameters, modulate the holographic growth and observation environment, and ensure the comprehensiveness and continuity of the biological rhythm scientific research. The sample 14 culture module has a sterilization function, and adopts an ultraviolet sterilization mode to sterilize the internal environment of the system, so as to ensure the growth environment of the detected sample 14. Meanwhile, due to the sensitivity of the biorhythm to light and temperature, the culture module is required to have better sealing performance, and the influence of factors such as light, temperature and the like of the external environment on monitoring activities is isolated. The sample 14 culture module is structurally composed of 4 parts, namely a culture box body, a light control module, a temperature control module, a sterilization module and the like. The culture dish 8 carries the sample 14 to be monitored and maintains an environmental seal, providing advantages for the growth and imaging of the sample 14. The illumination control module is used for driving and adjusting illumination intensity and illumination time in the incubator. The temperature control module is used for controlling the environmental temperature in the incubator, the environmental temperature is increased and reduced through the temperature control device, and the dark room condition during fluorescence imaging is ensured by designing the light-proof curved air duct 5. The sterilization module adopts the mode of ultraviolet irradiation to carry out the sterilization of overall environment, based on preceding degree tunable ultraviolet lamp and driver design, provides aseptic good living environment for being surveyed sample 14.

As shown in fig. 3, the fluorescence imaging module is used for acquiring fluorescence signals, and the fluorescence signals emitted by the sample 14 (biological individual, tissue/cell) are acquired by means of imaging so as to capture changes indicating biological rhythm. The fluorescence imaging is composed of a lens module, an optical filter module, an imaging module and the like, and the functions of automatic focusing, depth of field control, fluorescence imaging and the like are realized. The multispectral microscopic imaging system is designed by adopting light splitting based on an optical filter module, the optical filter module consists of an excitation light optical filter module 11 and an emission light optical filter module 12, the excitation light optical filter module 11 is used for outputting emission light with different wavelengths, and meanwhile, the emission light optical filter with a specific wavelength is adopted to eliminate noise generated by chloroplast background fluorescence, so that the signal-to-noise ratio of data is greatly improved. The optical filter module is used for light splitting, so that the wavelength dimension of a spectral image can be directly analyzed, one waveband of an emission spectrum is collected, information of spatial resolution data can be obtained, and image information of the whole waveband is obtained through continuous collection. By adjusting the optical filter module, specific wavelength can be switched from 420nm to 730nm, besides the wavelength is adjustable, different half-height widths can be selected for each central wavelength, the light transmittance from the broadband light source can be increased by selecting the wide half-height width, and the narrow half-height width can only transmit light of a few wavelengths from the broadband light source, so that multispectral microscopic imaging can be completed. The project proposes to adopt a high-performance SCMOS13 camera as a signal detector, and a signal gain function is added. The function can not only obviously enhance the target signal, but also greatly shorten the imaging exposure time so as to realize the implementation and collection of bioluminescence or fluorescence signals. The SCMOS13 camera employs a fast frame transfer chip design and overall exhibits good long-term stability compared to other fast imaging techniques, being the best choice for real-time fast imaging. With SCMOS13, the user can both perform long exposure imaging and achieve high quantum efficiency, low noise fast dynamic imaging.

As shown in fig. 4, the signal processing module includes embedded hardware, a software system, and a control system, and is responsible for environmental control of the incubator, driving of the imaging module, image storage and analysis, and human-computer interaction functions of the sample 14. The signal processing module is used for controlling the whole machine, processing and fusing the fluorescence image, superposing the fluorescence image and the natural light entity image, positioning the fluorescence signal on a specific tissue or cell, analyzing the expression quantity of a corresponding gene according to the fluorescence intensity of the fluorescence image, fitting a series of data together to form a continuous visual data set, and realizing more visual and accurate analysis on the dynamic change of a biological rhythm system. The technical key of the multispectral imaging system is the processing of fluorescence images, while the background subtraction and spectral separation techniques are key techniques affecting the signal-to-noise ratio and sensitivity. The complex and scientific spectrum separation algorithm is the core of the technology for identifying the multicolor fluorescence, and the spectrum separation is carried out by adopting a large number of image-made spectrum templates, so that the autofluorescence noise can be effectively removed, various fluorescences can be identified, and more accurate quantitative analysis can be carried out.

In the aspect of power supply of the whole instrument, a long research period is often needed for monitoring the biorhythm, the change of temperature and illumination can cause the change of circadian rhythm, and in order to prevent sudden power failure events, the system is provided with an Uninterruptible Power Supply (UPS).

The utility model discloses have following technological innovation point:

1) the multifunctional automation can control the integrated technology of the culture module: the integration level of the culture module is improved, and equipment such as illumination, temperature control, ultraviolet sterilization, linkage devices, fluorescence imaging and the like is ensured to be orderly installed and normally run in a small space;

2) fluorescence signal analysis and dynamic digital fitting: dynamic fluorescence imaging and monitoring data analysis and fusion are carried out, and long-range monitoring of circadian rhythm is realized;

3) and (3) universal design: the controllability of the biological material culture module and the sensitivity of the fluorescence imaging module are improved, so that the biological material culture module is suitable for various biological materials.

The key technologies of the project comprise multispectral fluorescence imaging technology research, fluorescence imaging sensitivity/real-time guarantee technology, optical image reconstruction technology, various fluorescence report systems and the like.

The above examples are merely illustrative of the present invention and do not limit the scope of the present invention, and all designs identical or similar to the present invention are within the scope of the present invention.

Claims

1. A material culture device suitable for studying biological rhythms, comprising: including the sealed chamber, the bottom of sealed chamber is equipped with the rotatable objective table that can place a plurality of culture dishes, and the culture dish is used for cultivateing biological individual sample or cell tissue sample, the top of sealed chamber is equipped with illumination light incident port, exciting light incident port and the imaging lens who is used for illumination and vegetation, the objective table is located in the application range of illumination light incident port, exciting light incident port and imaging lens, lay the culture dish on the objective table and be used for bearing and monitor the sample, the side is equipped with into wind gap fan and air outlet fan respectively about the sealed chamber.

2. A material culture device adapted to study biorhythms according to claim 1, wherein: the top of sealed chamber still is equipped with the ultraviolet lamp sterilizer, exciting light entrance, the illumination light entrance that is used for illumination and vegetation and ultraviolet lamp sterilizer follow sealed chamber arranges in proper order to the next door from the centre.

3. A material culture device adapted to study biorhythms according to claim 1, wherein: the outsides of air inlet fan and air outlet fan all are equipped with the bent shape wind channel that is used for the light-resistant.

4. A material culture device adapted to study biorhythms according to claim 1, wherein: the illumination light entrance ports are symmetrically distributed along the central line of the closed chamber.

5. A material culture device adapted to study biorhythms according to claim 1, wherein: the exciting light entrance ports are symmetrically distributed along the central line of the closed chamber.

6. A material culture device adapted to study biorhythms according to claim 1, wherein: and a heater is also arranged in the closed chamber.

7. A material culture device adapted to study biorhythms according to claim 1, wherein: and a xenon lamp light source is arranged at the exciting light incidence opening.

8. A material culture device adapted to study biorhythms according to claim 1, wherein: the culture dish includes a plurality ofly, the bottom of airtight chamber is equipped with rotatable objective table, each the culture dish evenly arrange in on the rotatable objective table.

9. A material culture system adapted to study biological rhythms, comprising a signal processing module, characterized in that: the material culture device suitable for studying biological rhythm of any one of claims 1 to 8, wherein the signal processing module is used for processing the illumination light signal, the excitation light signal and the imaging lens signal of the material culture device.