CN111778160A - Cell culture device - Google Patents
Cell culture device Download PDFInfo
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- CN111778160A CN111778160A CN202010708740.6A CN202010708740A CN111778160A CN 111778160 A CN111778160 A CN 111778160A CN 202010708740 A CN202010708740 A CN 202010708740A CN 111778160 A CN111778160 A CN 111778160A
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- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
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- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
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- C12M41/06—Means for regulation, monitoring, measurement or control, e.g. flow regulation of illumination
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Abstract
The invention provides a cell culture device, and belongs to the technical field of medical instruments. The cell culture apparatus includes: shell and setting are at inside cultivation board and the light source of shining of shell, the cultivation board is arranged a plurality of cultivation holes that set up on the cultivation base including cultivateing base and array, the cultivation hole is used for cultivateing the in vitro cell or the microorganism that set up respectively, the light source of shining includes the light source mounting panel and at least one light source that sets up on the light source mounting panel, the light source mounting panel sets up on the shell inner wall, a plurality of cultivation hole directions light-emitting of the last one line that corresponds of cultivation board of light source orientation, light source external power supply control work. The invention can research the biological mechanism of the isolated cells or the microorganisms in the culture process with the irradiation light source by taking the isolated cells or the microorganisms as an experimental means, thereby realizing the practical value of the cell culture or the microorganism culture in the biological engineering.
Description
Technical Field
The invention relates to the technical field of medical instruments, in particular to a cell culture device.
Background
Cell culture refers to all in vitro cultures, and means that cells are taken out from a living organism and incubated under specific in vivo conditions such as a simulated physiological environment in vivo to survive and grow. Cell culture work is widely applied to various fields such as biology, medicine, new drug research and development and becomes one of the most important basic sciences.
A large number of scientific studies prove that: the vegetative growth and the accumulation of secondary metabolites of autotrophic biological cells are directionally regulated by the wavelength under the irradiation of a light source with a certain wavelength, and the metabolic pathways of heterotrophic biological cells are also changed under the stimulation of the light source with a special wavelength, so that some bioactive substances are accumulated. Because light sources with different wavelengths have certain energy and can only be absorbed by certain special photosensitizing pigments or other photosensitizing substances (such as certain enzyme or non-enzyme protein receptors) in living cells, gene expression or corresponding metabolic networks are changed, and the expression quantity of certain specific secondary metabolites is increased. By utilizing the characteristic of living cells, the regulation and control mechanism of different wavelength light sources to tissue cells or cultured cells can be systematically and deeply researched, and the unknown light reaction mechanism is the important content of biological light reaction scientific research.
Light irradiation with various wavelengths has a certain photochemical effect on a complex metabolic network in a cultured cell, so that the light irradiation has a regulation function on signal transduction, enzyme activity and gene expression, and the activity of the cultured cell is influenced, and the light irradiation has great significance for the bioengineering industry which takes cell culture as a means to research the cell activity maintenance of the cultured cell for a long time. But no scientific instrument is available to meet the needs of this research.
Disclosure of Invention
The invention aims to provide a cell culture device which can research the biological mechanism of isolated cells or microorganisms in the culture process with an irradiation light source by taking the isolated cells or microorganisms as an experimental means, thereby realizing the practical value of the cell culture or the microorganism culture in biological engineering.
The embodiment of the invention is realized by the following steps:
an embodiment of the present invention provides a cell culture apparatus, including: shell and setting are at inside cultivation board and the light source of shining of shell, the cultivation board is arranged a plurality of cultivation holes that set up on the cultivation base including cultivateing base and array, the cultivation hole is used for cultivateing the in vitro cell or the microorganism that set up respectively, the light source of shining includes the light source mounting panel and at least one light source that sets up on the light source mounting panel, the light source mounting panel sets up on the shell inner wall, a plurality of cultivation hole directions light-emitting of the last one line that corresponds of cultivation board of light source orientation, light source external power supply control work.
Optionally, the light source is a linear light source, a light blocking partition is further disposed between adjacent rows of the culture wells, and the light blocking partition is disposed between the culture plate and the irradiation light source and used for blocking the irradiation light source from exiting to the outside of the plurality of culture wells in the corresponding row.
Optionally, one end of the light-blocking partition plate is fixedly arranged with the light source mounting plate, and the other end of the light-blocking partition plate is fixedly arranged with the culture base.
Alternatively, the culture plate is a 96-well plate, or the culture plate is a 24-well plate, or any multi-well plate.
Optionally, the light source is a laser, or the light source is an LED light source, the adjustable range of the emitting wavelength of the light source is 200-10000nm, and the adjustable range of the emitting power of the light source is 1-500mw/cm2。
Optionally, the light source is an LED light source, the light source is a plurality of LED lamp beads, the plurality of LED lamp beads are sequentially arranged on the light source mounting plate along the same linear direction, and a printed circuit is further formed on the light source mounting plate to control each LED lamp bead to work.
Optionally, the housing is opaque.
Optionally, the housing is made of opaque material.
Optionally, the culture body is a culture plate, and a plurality of culture holes are uniformly distributed on the culture plate.
Optionally, the culture device further comprises a control switch connected to the light source mounting plate, the control switch is connected to the printed circuit, and the control switch is used for controlling whether the light source is turned on or not.
Optionally, a heat dissipation structure is further disposed on the light source mounting board.
The embodiment of the invention has the beneficial effects that:
the cell culture device provided by the embodiment of the invention comprises a shell, a culture plate and an irradiation light source, wherein the culture plate and the irradiation light source are arranged in the shell, the culture plate comprises a culture base and a plurality of culture holes which are arranged on the culture base in an array mode, the culture plate is used for culturing in vitro cells or microorganisms, the culture plate can be a 96-hole plate or a 24-hole plate, the culture holes are used for culturing the in vitro cells or the microorganisms which are respectively arranged, each culture hole can be arranged into different culture environments, and experimental exploration is respectively carried out on the in vitro cells or the microorganisms. The wavelength of the irradiation light source is adjustable, the irradiation light sources with different wavelengths have certain photochemical effect on a complex metabolic network in an isolated cell or a microorganism, so that the regulation function on signal transduction, enzyme activity and gene expression is realized, and the biological mechanism of culturing the cell or the microorganism is further influenced. Therefore, the isolated cells or the microorganisms in the culture holes can be irradiated, the isolated cell culture or the microorganism culture is taken as an experimental means, the biological mechanism of the isolated cells or the microorganisms in the culture process with the irradiation light source is further researched, and the practical value of the cell culture or the microorganism culture in the biological engineering is realized.
Drawings
In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings needed to be used in the embodiments will be briefly described below, it should be understood that the following drawings only illustrate some embodiments of the present invention and therefore should not be considered as limiting the scope, and for those skilled in the art, other related drawings can be obtained according to the drawings without inventive efforts.
FIG. 1 is a schematic structural view of a cell culture apparatus according to an embodiment of the present invention;
FIG. 2 is a schematic structural diagram of a cell culture plate and an irradiation light source in the cell culture apparatus according to the embodiment of the present invention;
FIG. 3 is a schematic diagram of a cell culture plate of the cell culture apparatus according to the embodiment of the present invention;
FIG. 4 is a schematic diagram of a cell culture apparatus provided in an embodiment of the present invention, in which a light source is an LED lamp bead;
fig. 5 is a schematic diagram of a cell culture apparatus provided in an embodiment of the invention, in which a light source is an LED strip lamp.
Icon: 100-a housing; 110-a chute; 200-culture plate; 203-light blocking spacers; 210-a culture base; 220-culture well; 310-a light source mounting plate; 321-LED lamp beads; 322-LED strip lamp.
Detailed Description
In order to make the objects, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are some, but not all, embodiments of the present invention. The components of embodiments of the present invention generally described and illustrated in the figures herein may be arranged and designed in a wide variety of different configurations.
Thus, the following detailed description of the embodiments of the present invention, presented in the figures, is not intended to limit the scope of the invention, as claimed, but is merely representative of selected embodiments of the invention. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
In the description of the present invention, it should also be noted that, unless otherwise explicitly specified or limited, the terms "disposed," "mounted," "connected," and "connected" are to be construed broadly and may, for example, be fixedly connected, detachably connected, or integrally connected; can be mechanically or electrically connected; they may be connected directly or indirectly through intervening media, or they may be interconnected between two elements. The specific meanings of the above terms in the present invention can be understood in specific cases to those skilled in the art.
Fig. 1 is a schematic structural diagram of a cell culture apparatus provided in the present invention, and referring to fig. 1, an embodiment of the present invention provides a cell culture apparatus, including: the shell 100 and set up at the inside culture plate 200 and the light source that shines of shell 100, culture plate 200 arranges a plurality of culture holes 220 that set up on culture base 210 including culture base 210 and array, culture hole 220 is used for cultivateing the separation cell or the microorganism that set up respectively, the light source that shines includes light source mounting panel 310 and at least one light source that sets up on light source mounting panel 310, light source mounting panel 310 sets up on the shell 100 inner wall, the light source is towards a plurality of culture holes 220 directions light-emitting of the corresponding one line on culture plate 200, light source external power supply control work.
The irradiation light source is a low-intensity laser (also called non-intensity laser, also called weak laser), that is, a laser (also called weak laser or low-power laser low-energy laser) that does not directly cause irreversible damage to a biological tissue when the tissue is irradiated by the laser, and the low-intensity laser is different from the strong laser and is characterized by low output energy and no photothermal effect. The local temperature rise caused by low-intensity laser irradiation is not more than 0.1-0.75 ℃, and the biological change of local tissues cannot be caused. Low intensity laser is generally considered to mean a power output to the treatment site of 1-300mw/cm2The energy density is 1-4J/cm2. The low-intensity laser is adopted to continuously irradiate the isolated cells, so that the isolated cells can have various stimulation effects on the biological cells, including promoting cellular immune response, enhancing phagocytic capacity of macrophages, improving deformability of red blood cells, promoting cell proliferation and the like, and the activity of the irradiated isolated cells is improved to a certain extent. Among them, for example, some animal cells can increase the synthesis of Adenosine triphosphate (Adenosine triphosphate) under continuous irradiation of low-intensity laser, wherein Adenosine Triphosphate (ATP), which is a kind of coenzyme, has the effect of improving body metabolism, and is involved in the metabolism of body fat, protein, sugar, nucleic acid and nucleotide. Meanwhile, the adenosine triphosphate is also the main source of energy in vivo, and when energy is needed for absorption, secretion, muscle contraction, biochemical synthesis reaction and the like in vivo, adenosine triphosphate is decomposed into adenosine diphosphate and phosphate, and energy is released at the same time, so that human fibroblasts can be extracted under the continuous irradiation of low-intensity laserHigh human fibroblast activity. The low-intensity laser can comprise He-Ne laser (with the wavelength of 632.8nm), GaAlAs laser (with the wavelength of 820nm and 830nm), GaAs laser (with the wavelength of 904nm), Nd-YAG laser (with the wavelength of 1064ni) and the like, different types of laser are selected, the corresponding laser for generating the laser is different, and the selection can be carried out by a person skilled in the art according to the actual needs.
There are also a number of scientific studies that demonstrate: the vegetative growth and the accumulation of secondary metabolites of autotrophic biological cells are directionally regulated by the wavelength under the irradiation of a light source with a certain wavelength, and the metabolic pathways of heterotrophic biological cells are also changed under the stimulation of the light source with a special wavelength, so that some bioactive substances are accumulated. Because light sources with different wavelengths have certain energy and can only be absorbed by certain special photosensitizing pigments or other photosensitizing substances (such as certain enzyme or non-enzyme protein receptors) in living cells, gene expression or corresponding metabolic networks are changed, and the expression quantity of certain specific secondary metabolites is increased or reduced. Based on this property of biological cells, the regulation mechanism of different wavelength irradiation light sources to tissue cells, cells or microorganisms in vitro can be systematically and deeply studied, for example, the weak laser irradiation of microorganisms produces biostimulation effect, which can cause some change of biological function and can cause the variation of biological genetic property.
Weak laser irradiation of cells or microorganisms in vitro, regardless of the wavelength, causes a certain degree of acceleration of cell movement. This is because the low-intensity laser irradiation affects the exchange of calcium ions in the cell membrane, and the calcium ions can be concentrated in the cytoplasm in a short time. At the same time, the weak laser also affects Na+/K+And the conduction system is involved in regulating the calcium ion level, and consequently, the cell movement is accelerated.
Weak laser irradiation of cells and microorganisms ex vivo also increases a variety of enzymatic activities, including carbohydrate metabolism and mitochondrial respiratory chain important enzymes such as succinate dehydrogenase, cytochrome oxidase, NADPH oxidase, phosphorylase, and the like. Activation of these enzymes can increase endogenous insulin levels, promote sugar utilization and ATP production, and restore membrane Na+-K + -ATPase, regulationThe function of the ion pump is saved, and the ion balance and the membrane potential inside and outside the membrane are restored, so that the sugar metabolic acidosis, the polyol energy passage and the electrolyte disorder are corrected.
For example, referring to fig. 1, the light source is a linear light source, a plurality of pairs of sliding grooves 110 are arranged in parallel in the housing 100 of the culture apparatus, a certain height difference exists between the plurality of pairs of sliding grooves 110, two opposite side edges of the light source mounting plate 310 are inserted into the sliding grooves 110 to fix the linear light source, the light source mounting plate 310 is inserted into the sliding grooves 110 with different heights, which indicates that the light paths of the irradiation light source to the culture plate 200 are different, and can be used to study the influence of different irradiation light paths on the in-vitro cells or microorganisms during the culture process.
Illustratively, the culture plate is a 24-well plate, and the culture wells 220 in the 24-well plate are arranged in 4 rows and 6 columns, and since each linear light source with adjustable wavelength corresponds to the row/list of light facing the culture wells 220, there are various ways to realize wavelength adjustment, for example, adjustment of a single wavelength, or adjustment of continuous wavelengths, etc., and a person skilled in the art should select a specific adjustment way to adjust according to actual needs to study the influence of adjusting the wavelength of the light source in different ways on the culture object. Therefore, the number of the linear light sources arranged on the light source mounting plate 310 may be 4 or 6, when the number of the linear light sources arranged on the light source mounting plate 310 is 4, 4 linear light sources are arranged to respectively correspond to each row of the irradiation culture holes 220, and the emergent light of each linear light source cannot interfere with the culture holes 220 of the adjacent row, a special culture plate 200 needs to be arranged, or a special light source mounting plate 310 needs to be arranged, for example, each row of the culture holes 220 of the culture plate 200 has a light blocking partition plate 203, so that each linear light source can only correspondingly irradiate one row of the culture holes 220. The wavelengths of the 4 linear light sources are different from each other, and the same isolated cells or microorganism culture solution is arranged in the culture plate 200, so that the influence factors of the isolated cells or microorganisms in the culture process are only the parameters related to the light sources: the wavelength, the irradiation power density, the irradiation energy, the irradiation time and frequency, etc., i.e., the linear light sources with different wavelengths, are used to observe the influence of the parameters related to the different irradiation light sources on the biological mechanism of each line of the isolated cells or microorganisms, for example, the number of dead cells or microorganisms in the isolated cells or microorganisms in the culture apparatus is the least when the wavelength of the linear light source is observed (other irradiation parameters are consistent). Similarly, if the number of the linear light sources is set to 6, the in vitro cells or microorganisms in each row of the culture wells 220 in the 24-well plate can be selected and studied.
First, the housing 100 of the cell culture apparatus is made of a transparent material, which is convenient for an experimenter to observe experimental results, for example, the housing 100 is made of glass.
Second, the culture plate 200 is not limited to the 24-well plate described above, but may be other number of plates, and the culture plate 200 of the appropriate number of plates may be selected by those skilled in the art according to the isolated cells or microorganisms and the experimental requirements. The culture plate 200 in the present application may be transparent or opaque, for example, the culture plate 200 is black, and the black culture plate 200 has the function of blocking and absorbing light, so that the different illumination light between two adjacent well plates is not interfered with each other, which is only an example, and a person skilled in the art can select according to actual needs as long as the culture of the cultured microorganism or the isolated cell can be realized.
Thirdly, the culture plate 200 in the present application may be a plurality of well plates, or culture bottles or culture dishes, wherein the structure, size, shape and arrangement of the culture bottles and the culture dishes may be correspondingly arranged according to experimental needs, and the culture plates are not limited herein as long as the isolated cells or the microorganisms can be cultured under the irradiation light sources with different wavelengths, for example, each culture bottle or culture dish adopts one irradiation light source, and the wavelength of the irradiation light source can be correspondingly adjusted, so as to study the biological mechanism of the isolated cells or the microorganisms under the irradiation with different wavelengths.
Fourth, the length of time that the light source irradiates the cells or microorganisms ex vivo in the culture well 220 is adjustable in various ways, such as single adjustment, continuous adjustment, intermittent adjustment, pulsed adjustment, and random adjustment. Illustratively, the linear light source is adjusted by pulse type adjustment, and a corresponding pulse wave is input to the controller, so that the controller outputs a current according to the input pulse wave, thereby controlling the linear light source to irradiate the isolated cells or microorganisms in the culture hole 220 according to the input pulse wave control.
Fifthly, the light source may be a linear light source or a point light source, and the number of the light sources may be one or more, and different setting modes correspond to different experimental requirements, for example, when the same irradiation wavelength and irradiation power need to be controlled for one column or one row of the culture wells 220 on the culture plate 200, the setting of the linear light source is convenient for control. The above is only an example of one of the setting manners, and a person skilled in the art can set a certain number of light sources with corresponding shapes according to specific experimental requirements as long as the experimental requirements can be met and the biological mechanism of the isolated cells or the microorganisms in the culture process can be studied.
The cell culture device provided by the embodiment of the invention comprises a shell 100, a culture plate 200 and an irradiation light source, wherein the culture plate 200 is arranged inside the shell 100, the culture plate 200 comprises a culture base 210 and a plurality of culture holes 220 which are arranged on the culture base 210 in an array mode, the culture plate 200 is used for culturing in vitro cells or microorganisms, the culture plate 200 can be a 96-hole plate or a 24-hole plate, the culture holes 220 are used for culturing in vitro cells or microorganisms respectively, each culture hole 220 can be set to be in different culture environments, and experimental exploration is respectively carried out on in vitro cells or microorganisms. The wavelength of the irradiation light source is adjustable, the irradiation light source with different wavelengths has a certain photochemical effect on a complex metabolic network in vitro cells or microorganisms, so that the regulation function on signal transduction, enzyme activity and gene expression is realized, and the biological mechanism of a culture object is influenced. Therefore, the irradiation of the isolated cells or the microorganisms in the culture hole 220 can be realized, the isolated cells or the microorganisms in the culture hole 220 can be irradiated, the isolated cell culture or the microorganism culture is used as an experimental means, the biological mechanism of the isolated cells or the microorganisms in the culture process with the irradiation light source is further researched, and the practical value of the cell culture or the microorganism culture in the biological engineering is realized.
Fig. 2 is a schematic structural diagram of a culture plate and an irradiation light source in a cell culture apparatus according to an embodiment of the present invention, please refer to fig. 2, further, in order to eliminate the influence of light emitted from the light source on adjacent culture wells 220, in this embodiment, when the light source is a linear light source, a light blocking partition 203 is further disposed between adjacent rows/columns of the culture wells 220, and the light blocking partition 203 is disposed between the culture plate 200 and the irradiation light source for blocking the irradiation light source from being emitted out of the corresponding row of the plurality of culture wells 220.
It should be noted that the light blocking partition plates 203 oppositely arranged on the two sides of the linear light source can eliminate the influence of the emergent light of the linear light source on the adjacent linear light source, and the two sides of the light blocking partition plates 203 are both coated with the reflective material, which can improve the utilization rate of the emergent light of the linear light source.
Further, one end of the light blocking partition plate 203 is fixed to the light source mounting plate 310, and the other end is fixed to the culture base 210.
Illustratively, the width of the light-blocking partitions 203 is reduced at both ends, a groove is provided between the adjacent linear light sources in the light source mounting plate 310, the width of the groove corresponds to the width of the end of the light-blocking partitions 203, and similarly, a receiving groove having a width corresponding to the width of the end of the light-blocking partitions 203 is provided between the adjacent culture holes 220 in the culture base 210, so that one end of the light-blocking partitions 203 is inserted into the groove in the culture base 210 and the other end is inserted into the receiving groove in the culture base 210.
It should be noted that the grooves disposed on the light source mounting plate 310 may be arranged in an array, and similarly, the accommodating grooves disposed on the culture base 210 may also be arranged in an array, and those skilled in the art can insert the light blocking partitions 203 into the corresponding grooves and accommodating grooves according to the experimental requirements, so as to eliminate the interference of the linear light sources on the rows/columns in the adjacent culture holes 220.
Fig. 3 is a schematic structural diagram of a culture plate in a cell culture apparatus according to an embodiment of the present invention, where the culture plate 200 is shown above, and referring to fig. 3, the culture plate 200 is shown in this embodiment as a 96-well plate, or the culture plate 200 is a 24-well plate, or the culture plate 200 is any other multi-well plate.
For example, the cell culture device is used for exploring the influence of irradiation light sources with various wave bands on the irradiation of the ex-vivo cells or microorganisms, and corresponding experimental data under various experimental conditions are generally required to be carried out so as to carry out relevant comparative analysis, experimental analysis and conclusion on the influence of various experimental conditions on the ex-vivo cells or microorganisms. This requires that many experimental conditions of different dimensions cross each other, often requiring a large number of experiments and data records to support the analysis of the experiments and the drawing of conclusions. For example, to study the relationship of the activity of lymphocytes in human body being increased by weak laser irradiation, a plurality of samples of lymphocytes are sampled and respectively given to weak laser conditions with different wavelengths or different irradiation time periods, and then experimental data are recorded.
In the experimental process, if a large number of lymphocyte samples are inactivated or dead in the experimental process, on one hand, the effective experimental data amount can be greatly reduced, on the other hand, the experimental process and the experimental times are prolonged, and even in the process of increasing the experimental times, the experimental data error is larger due to the influence of other known or unknown experimental conditions, so that the accurate conclusion of the final experimental conclusion is influenced.
By the cell culture device, the sampled isolated cells or sampled microorganisms can be respectively and correspondingly placed in a 24-hole plate or a 96-hole plate, different irradiation experiment conditions are respectively given in one experiment process, the sampled isolated cells or sampled microorganisms in the cell culture device are irradiated by corresponding weak laser, and the biological mechanism of the sampled cells or sampled microorganisms in the experiment process is researched, so that the isolated cells or microorganisms irradiated by the weak laser are obtained, and the purpose of researching the biological mechanism of the isolated cells or microorganisms is achieved.
Under the above conditions, the culture plate 200 of the 96-well plate can be selected, the 96-well plate has 12 rows and 8 columns, in order to wait for experimental data of multiple groups of wavelengths, the wavelengths of the irradiation light sources corresponding to the linear light sources in each row of the 96-well plate are different, so that experimental data of 12 wavelength bands can be obtained, the number of the corresponding linear light sources of the irradiation light sources is also 12, the wavelength of each linear light source is adjusted, the isolated cells in the 96-well plate are cultured respectively, and the experimental data are recorded.
The cell culture apparatus may also be used to explore the biological impact of other elements on cells or microorganisms ex vivo. For example, the linear light sources in the irradiation light sources are all arranged at the same wavelength, and the culture solution in the culture wells 220 are arranged differently, so that the biological influence of the culture solution in the culture environment of the cells or the microorganisms on the cells or the microorganisms in vitro during the culture process can be researched.
It should be noted that the plate 200 may have different well sites selected according to the experimental requirements and the nature of the cells or microorganisms ex vivo. In this embodiment, a 96-well plate and a 24-well plate are specifically shown, and the isolated cell or microorganism culture apparatus is not only suitable for these two well plates, but also can be used in other numbers of well plates, and those skilled in the art can select a suitable culture plate 200 according to the characteristics of the isolated cell or microorganism to be cultured and the experimental requirements.
Further, the light source can be laser or LED light source, the adjustable range of the emitting wavelength of the light source is 200-10000nm, and the adjustable range of the emitting power of the light source is 1-500mw/cm2。
In order to further study the biological influence of the light source on the isolated cells or microorganisms, the emission wavelength and the emission power of the light source suitable for the isolated cells or microorganisms can be set according to different types of isolated cells or microorganisms, so that the influence of the light sources with different emission wavelengths and different emission powers on the biological mechanism of the isolated cells or microorganisms can be further studied.
A Light Emitting Diode (LED) is a solid-state light source having high luminance, high efficiency, and long life. LED light-emitting principle: the core part of the light emitting diode is a wafer consisting of a P-type semiconductor and an N-type semiconductor, and a transition layer called a P-N junction is arranged between the P-type semiconductor and the N-type semiconductor. In P-N junctions of some semiconductor materials, injected minority carriers and majority carriers when recombined release excess energy in the form of light, thereby converting electrical energy directly into light energy. The P-N junction is applied with reverse voltage, so that minority carriers are difficult to inject, and light is not emitted. When it is in forward working state (i.e. forward voltage is applied to two ends), when the current flows from anode to cathode of LED, the semiconductor crystal can emit light rays with different colors from ultraviolet to infrared, and the intensity of light is related to current.
The LED light source can emit laser light, the laser light emitted by the LED light source is generally low-intensity laser light, and the heat generated by the LED light source is small, so the LED light source is adopted as the irradiation light source in the present embodiment.
Research shows that under the action of low-intensity laser and an LED light source, the change of animal cells is mainly related to wavelength, irradiation dose and irradiation mode, and laser with better correlation is not a necessary condition; whereas LEDs have similar effects to low-energy lasers in terms of cellular protein synthesis rate, growth rate, enzyme activity, and CAMP (Cyclic Adenosine monophosphate) levels. Continuous irradiation with low-intensity laser light of 830nm wavelength can increase the synthesis of adenosine triphosphate, thereby increasing the activity of cells.
Illustratively, the low-intensity laser may include He-Nc laser (wavelength 632.8nm), GaAlAs laser (wavelength 820nm, 830nm), GaAs laser (wavelength 904nm), Nd: YAG laser (wavelength 1064nm), etc., different types of lasers are selected, and the corresponding lasers for generating the laser are different, and those skilled in the art can select the laser according to actual needs.
Fig. 4 is a schematic diagram of a cell culture apparatus provided in an embodiment of the present invention, in which the light source is an LED lamp bead 321, and further, referring to fig. 4, the light source is a plurality of LED lamp beads 321 sequentially arranged on a light source mounting board 310 along the same linear direction, and a printed circuit is further formed on the light source mounting board 310 to control each LED lamp bead 321 to work.
Illustratively, the cultivation board 200 is a 24-hole board, and the 24-hole board has 4 rows and 6 columns, the LED lamp beads 321 are sequentially arranged on the light source mounting board 310 along the same linear direction to form a plurality of linear light sources, if each linear light source corresponds to each row of the cultivation board 200, and in order to obtain a plurality of sets of experimental data, the wavelength of each LED lamp bead 321 can be adjusted and controlled to be different, and the LED lamp bead 321 in each linear light source corresponds to each row of the cultivation holes 220 in the 24-hole board, so that the wavelength of the light source irradiated by each cultivation hole 220 in the cultivation board 200 can be different, and thus, the number of experiments can be reduced, and the experiment efficiency can be improved.
Fig. 5 is a schematic diagram of the cell culture apparatus provided in the embodiment of the invention, in which the light source is the LED strip lamp 322, and the wavelength of the illumination light source for only controlling each row/column of the culture holes 220 of the culture plate 200 is the LED strip lamp 322, please refer to fig. 5, where each LED strip lamp 322 corresponds to each row/column of the culture holes 220 in the culture plate 200, and the LED strip lamp 322 is configured under the experimental condition that the complexity of the circuit structure can be reduced.
The LED light source is a novel light source in a laser family, belongs to the category of 'weak laser', and is a solid semiconductor device capable of converting electric energy into visible light, and the LED light source can directly convert the electric energy into light energy. After the biological tissue is irradiated by the laser, if the irreversible damage of the biological tissue is directly caused, the laser at the irradiated surface is called as strong laser; if the damage is not directly irreversible, it is called weak laser. The LED light source does not contain harmful substances such as mercury and lead, has no infrared and ultraviolet pollution, and does not generate pollution to the outside during use.
Generally, the LED adopts laser LED lamp bead chips with four basic colors of white (including various illumination white lights such as cold white and warm white), red, green and blue (R, G, B), the lamp bead chips are packaged in various forms, each group of colors can be separately used, and are respectively connected with a driving circuit and a singlechip. A user can control the red, green and blue (R, G, B) LED lamp bead chips to approximately adjust almost all the light colors visible to human eyes according to the principle of three primary colors (all colors can be mixed by the three primary colors of red, green and blue according to a certain proportion). The user can adjust the brightness of the chips of the white, red, green and blue (R, G, B) four-color lamp beads through a remote controller or a button connected with a wire on the lamp, thereby realizing the adjustment of the wavelength of the LED.
In this embodiment, the housing 100 is opaque.
It should be noted that, the housing 100 is configured to be opaque, and there are various specific configuration methods, for example, an outer surface of the housing 100 is coated with a layer of opaque material, or the housing 100 is made of an opaque material, as long as the housing 100 is made of an opaque material, and the purpose of configuring the housing 100 to be opaque includes two aspects, on one hand, the opaque housing 100 enables most of energy contained in laser emitted from the low-intensity laser source to act on in-vitro cells, so as to effectively reduce loss of energy of the laser source, and on the other hand, the opaque housing 100 can block interference of stray light in an external environment on photobiological action of in-vitro cells or microorganisms.
In this embodiment, the housing 100 is made of opaque material.
It should be noted that, in the first scheme, only the casing 100 is limited to be made of an opaque material, and the opaque material has multiple kinds, for example, the casing 100 is made of ceramic, or the outer surface of the casing 100 is coated with a layer of light-shielding material, in the second scheme, the inner surface of the casing 100 is provided with a reflective layer, and the inner surface of the casing 100 is provided with the reflective layer, which can be realized by multiple methods, for example, the casing 100 is made of glass with a metal reflective layer, or the inner surface of the casing 100 is plated with a layer of reflective material, in the third scheme, the casing 100 is made of an opaque material, and the inner surface of the casing 100 is provided with a reflective layer, which can be selected to be an opaque material, and the reflective layer is plated with a layer of reflective material, so as to improve the utilization rate of the culture plate 200.
Furthermore, the cell culture device also comprises a circuit structure arranged on the power supply mounting plate, the circuit structure comprises a plurality of wires which are arranged in parallel, one end of each wire is connected with the light source, the other end of each wire is connected with the control switch, the plurality of wires are used for transmitting the current required by the work of the light source, and the control switch is used for controlling the on/off of the light source.
The control switch is used to control and adjust the wavelength change of the irradiation light source and the time period for the irradiation light source to irradiate the isolated cells or microorganisms in the culture well 220.
In this embodiment, the irradiation light source is further provided with a heat dissipation structure.
It should be noted that the irradiation characteristics of the low-intensity laser source are low output energy and no photo-thermal effect, so that the irradiation light source does not generate excessive heat in the process of irradiating the isolated cells, but the irradiation light source is started to emit low-intensity laser in the closed environment of the housing 100 for a long time, which is difficult to avoid causing the environment in the housing 100 to generate certain heat, so that the temperature in the housing 100 is increased, thereby affecting the temperature of the culture environment, the isolated cells or microorganisms are particularly sensitive to the temperature change in the culture environment, and the temperature change affects the culture effect of the isolated cells or microorganisms, therefore, it is necessary to provide a heat dissipation structure on the irradiation light source to ensure that the temperature of the isolated cells or microorganisms culture environment is kept constant.
According to the cell culture device provided by the embodiment of the invention, the culture plate 200 is irradiated by the light-tight shell 100 and the irradiation light source, wherein the irradiation light source can be a linear light source consisting of LED lamp beads or an LED strip-shaped lamp 322. The light blocking partition plate 203 is arranged between the irradiation light source and the culture plate 200 and used for blocking light beams emitted by the irradiation light source to the rows/columns of the adjacent culture holes 220, so that experiments of multiple groups of irradiation light sources can be carried out simultaneously, the diversity of cell or microorganism culture is improved, and the biological influence of the cell or microorganism culture under different culture environments can be researched by taking the cell or microorganism culture as an experimental means.
The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention, and various modifications and changes may be made by those skilled in the art. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Claims (8)
1. A cell culture apparatus, comprising: the shell and setting are in inside cultivation board and the irradiation source of shell, the cultivation board is arranged the setting including cultivateing base and array and is in a plurality of cultivation holes on the cultivation base, the cultivation hole is used for cultivateing the separation cell or the microorganism that set up respectively, irradiation source includes the light source mounting panel and at least one light source that sets up on the light source mounting panel, the light source mounting panel set up in on the shell inner wall, the light source orientation cultivate on the board corresponding a plurality of on a line the direction light-emitting of cultivation hole.
2. The cell culture apparatus of claim 1, wherein the light source is a linear light source, and a light blocking partition is disposed between adjacent rows of the culture wells and between the culture plate and the illumination light source for blocking the linear light source from exiting the plurality of culture wells of a corresponding row.
3. The cell culture apparatus according to claim 2, wherein the light blocking partition has one end fixed to the light source mounting plate and the other end fixed to the culture base.
4. The cell culture apparatus of claim 1, wherein the culture plate is a 96-well plate, or wherein the culture plate is a 24-well plate.
5. The cell culture apparatus as claimed in claim 1, wherein the light source is a laser, or the light source is an LED light source, the adjustable range of the emitting wavelength of the light source is 200-10000nm, and the adjustable range of the emitting power of the light source is 1-500mw/cm2。
6. The cell culture device according to claim 5, wherein the light source is an LED light source, the LED light source is a plurality of LED beads, the LED beads are sequentially arranged on the light source mounting plate along the same linear direction, and a printed circuit is further formed on the light source mounting plate to control each LED bead to operate.
7. The cell culture assembly of claim 1 wherein the housing is light-impermeable.
8. The cell culture apparatus of claim 1, wherein the light source mounting plate is further provided with a heat dissipation structure.
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