WO2013086915A1

WO2013086915A1 - Biological culturing device with complete wavelength light-control

Info

Publication number: WO2013086915A1
Application number: PCT/CN2012/084420
Authority: WO
Inventors: 董静洲; 王瑛; 刘春朝; 徐信兰
Original assignee: 中国科学院华南植物园
Priority date: 2011-12-16
Filing date: 2012-11-10
Publication date: 2013-06-20
Also published as: CN102433259A; CN103384720A; CN102433259B; CN103384720B

Abstract

A biological culturing device with complete wavelength light-control comprises a box body (12) which is provided with a heat insulating layer on the outside, light wavelength controllable LED illumination panels which are arranged on the inner top side and four lateral sides of the box body (12), a temperature and humidity sensor which is arranged in the box body (12), a heater (7a), a humidifier (8) and a miniature mute compressor (7b) which are fixed on the bottom of the box body (12). Biologic photo-reaction researches can be performed in plant tissue culture, cell culture, microbiological culture and animal culture ways in the culturing device provided with complete wavelength light-control and quartz culture flask (13). The operation is simple. The light source can be subjected to wavelength combination and illumination time program setting according to different requirements.

Description

The invention relates to the field of biological culture, in particular to a plant material (including callus and seedling) culture device for a full-wavelength controllable light source, which is suitable for illumination control of biological cells or tissues. Or plant photoreaction studies, physiological metabolism studies or animal photoreaction physiology studies.

Background

A large number of scientific studies have proved that the vegetative growth and secondary metabolite accumulation of autotrophic biological cells under the irradiation of a certain wavelength of light source are regulated by the wavelength, and the metabolic pathway of the heterotrophic biological cells stimulated by the special wavelength light source will also change. Lead to the accumulation of some biologically active substances. This feature of living cells can be used to industrially produce biologically active substances or drugs that humans need. Because different wavelengths of light source have a certain amount of energy, and can only be absorbed by some special phytochrome or other photosensitive substances (such as some enzymes or non-enzymatic protein receptors) in living cells to make gene expression or corresponding metabolic network. A change has occurred, thereby causing an increase in the expression level of certain specific secondary metabolites. This is the theoretical basis of the apparatus of the present invention. By utilizing this characteristic of living cells, it is possible to systematically and in-depth study of the physiological activities of different wavelengths of light on organisms or tissue cells, and the regulation mechanism of cultured cellular metabolic networks. These unknown photoreaction mechanisms are important for scientific research of biophotoresponse.

Light irradiation at various wavelengths has a certain photochemical effect on plant growth and differentiation, metabolism, physiological activities of animals, and complex metabolic networks in cultured cells, thereby regulating signal transduction, enzyme activity, and gene expression. The bioengineering industry for the purpose of biophotometric reaction theory and cell culture as a means of producing bioactive substances and drugs is of great significance. Especially the tissue culture or cultivation of rare medicinal plants or the study of cell fermentation photoreaction is of great significance, but there is no scientific instrument that meets the needs of this research.

Summary of the invention

The object of the present invention is to solve the above problems in the prior art, and to provide a biological growth device for a full-wavelength controllable light source, which provides a full-wavelength light source and an LED fluorescent light source ranging from 200 nm to 1000 nm. According to the needs of scientific research, the light source controller on the device can select light of any wavelength in the wavelength range to combine to obtain a mixed light source or a monochromatic light source with a special wavelength combination, and use the light source to ferment cells or plant call tissues. Or plant seedlings or small animals for photoreaction studies. Especially suitable for: the regulation of the expression of certain genes by specific wavelengths of light, the regulation of the accumulation of secondary metabolites, the regulation of plant cell growth and differentiation, the regulation of physiological activities and metabolism of plants, The physiological activities of small animals. The object of the invention can be achieved by the following technical solutions:

A biological culture device for a full-wavelength controllable light source, comprising a box body and a culture bottle, characterized in that:

In the box:

The first side stereo LED lighting panel controller, the second side stereo LED lighting panel controller, the third side stereo LED lighting panel controller, the fourth side stereo LED lighting panel controller, the fifth side stereo LED lighting panel controller, The temperature, humidity, and UV lamp controllers are attached to the inside of the top panel of the cabinet, arranged from left to right.

The first stereo LED illumination panel controller is connected to the second stereo LED illumination panel controller, the second stereo LED illumination panel controller is connected to the third stereo LED illumination panel controller, and the third stereo LED illumination panel control The fourth side stereo LED illumination panel controller is connected with the fourth side stereo LED illumination panel controller, the fifth side stereo LED illumination panel controller and the temperature, humidity and ultraviolet lamps are connected. The controller is connected; the digital display and parameter setting panels of each controller are exposed and located at their respective positions above the box door.

The three components of the heater, the compressor and the humidifier are respectively fixed on the bottom plate of the casing.

The LED light source panel is fixed on the inner wall of the corresponding cabinet panel; the inner wall panel (quartz panel) is covered on the LED light source panel; the other four inner walls of the cabinet are arranged in the same manner.

The first valve is fixed above the top panel of the box, the vent hole of the first valve penetrates the top of the box at a vertical portion to facilitate ventilation; the second valve is a vent valve located on the bottom plate of the box, and the second valve also penetrates the box at a vertical portion At the bottom of the body, the first valve and the second valve are used for ventilation in the tank.

The first box door is fixed on the panel on the left side of the box body, the second box door is fixed on the panel on the right side of the box body, the two box doors are opposite; the first box board is installed in the box body at a height of one-half, The second box plate is installed in the vertical center of the box body in the left and right direction.

The culture bottle includes a quartz bottle body, a quartz bottle cap, a sterile vent plug socket, a quartz bottle body height of 5-20 cm, and a diameter of 3-20 cm. It can also be customized according to research needs. The inside of the quartz bottle cap is a grinding mouth and a cover. On the mouth of the bottle, the outer surface of the bottle mouth is also ground, which is easy to seal with the quartz cap. The aseptic vent plug is located at the top of the quartz cap. The top of the quartz cap has an opening under the sterile vent plug for easy insertion. Keep sterile air into the bottle after the stopper.

The inner wall panel is a quartz panel.

The LED light source board is provided with an LED lamp and an LED fluorescent lamp with a wavelength range of 200-1000 nm, wherein the wavelength range is 200-1000 nm, the LED lamp is used to adjust the light source of different wavelengths, and the LED fluorescent lamp is used for the comparison of the traditional light source, the full wavelength The minimum gradient of the wavelength adjustment of the LED device is 5-10 nm.

A biological device for a full-wavelength controllable light source, which comprises a peripheral box, a quartz bottle body, a quartz bottle cap, a sterile vent plug socket, a first-sided stereo LED illumination panel controller, and a second-sided stereo LED illumination panel control Third dimension LED lighting panel controller, fourth-sided stereo LED lighting panel controller, fifth-sided stereo LED lighting panel controller, temperature, humidity and UV lamp controller, heater and compressor, humidifier, quartz panel, LED light, Ventilation valve, box door, box board. The connection relationship is as follows: The outer casing is a heat-insulating plate made of foam insulation material in the middle of the double-layer stainless steel panel, and the three sides of the outer casing are equipped with LED panels (electronic industrial circuit boards), LED lamp beads. Fixed to the LED panel. The LED beads on each board are closely arranged in the order of horizontal or vertical according to the wavelength of the lamp, ie: 200nm, 205nm, 21 Onm 1000nm, 200nm, 205nm, 21 Onm 1 OOOnm and LED fluorescent lamps, and so on. Arranged to the end of the circuit board, one of the boards is arranged in the opposite direction to the opposite side of the board to ensure the uniformity of the light source of the fermenting cells, tissues or plants in the box, and the control of the five LED panels The first surface is located below the top of the box, the first side of the stereo LED lighting panel controller, the second side of the stereo LED lighting panel controller, the third side of the stereo LED lighting panel controller, the fourth side of the stereo LED lighting panel controller, the fifth side A stereo LED lighting panel controller (shown in Figure 1), in addition, a temperature, humidity and UV lamp controller 6 located below the top of the cabinet is connected to the heater and the mute compressor and humidifier. The LED panel is located between the peripheral box and the quartz glass plate inside the box. The front, rear, left and right and the top quartz glass plates in the box are tightly combined with the stainless steel plate at the bottom of the box to form a closed box space. The top quartz glass plate, the top LED panel and the top panel of the cabinet have a small hole connected to the venting valve, and the bottom plate has a small hole of the same size to ensure air circulation.

A biological device for a full-wavelength controllable light source, comprising a peripheral box, an LED illuminating plate disposed in a peripheral box body, an inner wall composed of a quartz glass plate in the box body, and a top, front, rear, left and right LED illuminating panel The controller is a first-sided stereo LED illumination panel controller, a second-sided stereo LED illumination panel controller, a third-sided stereo LED illumination panel controller, a fourth-sided stereo LED illumination panel controller, and a fifth-sided stereo LED illumination. Panel controller, temperature, humidity and UV lamp controllers are temperature and humidity controllers and germicidal light switch buttons, heaters and compressors for heating and cooling systems (including a heater and a micro-quiet compressor, and temperature sensor), The humidifier is a humidifier. The heater and compressor and humidifier are digitally controlled by temperature, humidity and UV lamp controllers. The venting valve is an air convection hole. It is characterized by: providing space for five-sided illumination, which can be used for tissue culture, cell culture, microbial culture, Plant seedling culture and animal culture; five-sided LED light source is controlled by controller, five controllers have wavelength selection and combination function in the range of 200-1000 nm and LED fluorescent lamp control function, illumination time program setting function; They are made up of quartz glass panels, smooth and sterile; the silent compressor ensures quiet inside the box; the humidifier adjusts the humidity inside the box to facilitate the cultivation of plant seedlings.

Used in conjunction with this incubator is a quartz culture flask for tissue culture or cell culture. The bottle body is made of quartz glass to ensure the permeability of all wavelengths in the range of 200-1000 nm. It is the bottle cap, the bottle mouth, the contact between the cap and the bottle mouth is frosted contact, easy to seal, and it is sterile ventilation. Plug the insertion port. It is characterized in that: the entire bottle is made of quartz glass, The light wave in the range of 200-1000 nm has good permeability and is suitable for full-wavelength biophotoresponse research. The cap has a matte port and a vent plug, which solves the problem of ventilation in tissue culture or cell culture, and simplifies the sterility experiment. operating.

The full-wavelength LED device has a wavelength range of 200-1000 nm and an LED fluorescent lamp, and the minimum gradient of the wavelength adjustment of the full-wavelength LED device is 5-10 nm.

The first three-dimensional LED illumination panel controller, the second stereo LED illumination panel controller, the third stereo LED illumination panel controller, the fourth stereo LED illumination panel controller, and the fifth stereo LED illumination panel controller respectively It is connected to the top, front, rear, left and right LED panels; the heating and cooling devices and humidifiers of the compressor and heater including the mute function are connected to the controller and controlled by the controller.

The full-wavelength LED device has a wavelength range of 200-1000 nm, and the wavelength-adjusted minimum gradient of the full-wavelength LED device is 5-10 nm. The LED lamp bead is fixed on the circuit board. The LED beads on each board are closely arranged, and the order is arranged horizontally or vertically according to the wavelength of the lamp bead, ie:

200nm, 205nm, 21 Onm 1000nm, 200nm, 205nm, 21 Onm 1 OOOnm and LED fluorescent lamps, and so on, to the end of the board, one of the boards is arranged in the opposite direction to the opposite board, To ensure the uniformity of the light source of the fermented cells, tissues or plant seedlings in the box, the control function of each light source controller is: a plurality of LED beads of the same wavelength on the LED panel controlled by the controller are connected in parallel as a power control unit .

The light-shielding plate can divide the box body into four or more culture spaces, and each of the culture spaces can respectively control the wavelength of the light source, and can simultaneously perform cell or tissue culture comparison of multiple wavelength light sources in the same incubator. test.

The utility model comprises a peripheral box body, an inner wall made of quartz glass disposed in the outer box body and a light blocking plate (the light blocking plate can be added, reduced or removed as needed), and the five-dimensional LED panel in the box body provides a stereo light source, which is arranged in the box Heater, compressor and humidifier at the bottom of the body and connected to the top controller.

The full-wavelength LED device as described above has a wavelength range of 200-1000 nm and an LED fluorescent lamp, and the minimum gradient of the wavelength adjustment of the full-wavelength LED device is 5-10 nm.

The device also includes a venting valve at the top of the tank to maintain the venting inside the box, and a ventilating valve has a micro ventilating fan. Compared with the prior art, the beneficial effects of the present invention are:

1. Simple structure, simple and convenient operation;

2. Provide accurate light source control system for scientific research needs.

3. Add a full-wavelength controllable light source to the existing incubator to fill the gaps in the instruments and techniques required for biophotonic reaction research.

4. The inner wall of the tank and the culture bottle are made of quartz glass to ensure the transmittance of the light source of each wavelength.

5. According to the needs of the fermenting cells and the target product, the light source is changed into a light source plate of a specific wavelength combination, which can be used for work. Industrialized tissue culture, using the principle of photoreaction to improve the growth rate of tissue cells, the amount of growth and the production of the target product

FIG. 1 is a schematic structural view of a biological culture apparatus for a full-wavelength controllable light source.

Figure 2 is a schematic view of the structure of a quartz bottle body.

Among them: 1- first stereo LED lighting panel controller, 2- second stereo LED lighting panel controller, 3-third stereo LED lighting panel controller, 4-fourth stereo LED lighting panel controller, 5-5th stereo LED lighting panel controller, 6-temperature, humidity and UV lamp controller, 12-box, 13-culture flask, 13a-quartz bottle, 13b- quartz cap, 13c-sterile ventilation Plug socket, 13d-bottle, 7a-heater, 7b-compressor, 8-humidifier, 9-inner wall panel (quartz panel), 10-LED light source panel, 11-first valve, 14a-first door , 14b - second box door, 15a - first box board, 15b - second box board, 16 - second valve.

Concrete

Example 1

As shown in Fig. 1, a biological culture device for a full-wavelength controllable light source comprises a casing 12 and a culture bottle 13. The first side stereo LED lighting panel controller 1, the second side stereo LED lighting panel controller 2, the third side stereo LED lighting panel controller 3, the fourth side stereo LED lighting panel controller 4, the fifth side stereo LED lighting The panel controller 5, the temperature, humidity and ultraviolet lamp controllers 6 are respectively fixed on the inner wall of the casing, and the digital display and parameter setting panels are fixed at the uppermost portion of the outer casing. The three components of the heater 7a, the compressor 7b and the humidifier 8 are respectively fixed on the bottom plate of the casing 12, and the inner wall panel (quartz panel) 9 is covered on the LED light source panel, and is fixed on the panel of the casing 12 by screws, respectively. The LED light source panel 10 is fixed on the panel of the corresponding cabinet 12, the ventilation valve 11 is fixed above the top panel of the cabinet 12, and the top panel has a vent hole connected to the valve. The quartz panel at the top of the box and the LED panel at the top of the box are both There is a venting hole and is connected to the valve vent for ventilation inside the box. The first box door 14a is fixed on the panel on the left side of the box body, the second box door 14b is fixed on the panel on the right side of the box body, and the two box doors are opposite; the first box board 15a is installed in the box. At a height, the second box plate 15b is mounted at a vertical center portion of the box in the left-right direction. The composition of the culture bottle 13 is: The height and diameter of the quartz bottle body 13a can be customized according to the research needs, the inside of the quartz bottle cap 13b is a grinding mouth, and the cover is 13d, and the outer surface of the bottle mouth 13d is also ground, which is convenient for the cap 13 The sealed, sterile vent plug socket 13c is located on top of the quartz bottle cap 13b, and the top of the quartz cap 13b has an opening under the sterile vent plug socket 13c to facilitate insertion of the sterile plug to maintain sterile air into the bottle.

The LED light source boards on each circuit board are closely arranged in the order of horizontal or vertical according to the wavelength of the lamp beads, namely: 200 nm, 205 nm, 210 nm 1000 nm, 200 nm, 205 nm, 21 Onm lOOOnm

And LED fluorescent lamps, and so on, are discharged to the end of the board, one of the boards is arranged in the opposite direction and opposite The arrangement of the circuit boards is reversed to ensure the uniformity of the light source of the fermenting cells, tissues or plant bodies in the box. The controllers of the five LED panels are respectively located on the first side of the box body, and the first side of the stereo LED illumination panel controller 1 is The second side stereo LED illumination panel controller 2, the third side stereo LED illumination panel controller 3, the fourth side stereo LED illumination panel controller 4, the fifth side stereo LED illumination panel controller 5 (shown in Figure 1) In addition, temperature and moderate monitoring and control of the temperature, humidity and UV lamp controller 6 located below the top of the cabinet, the temperature, humidity and UV lamp controller 6 are connected to the heater 7a, the compressor 7b and the humidifier 8. The LED light source panel 10 is located between the peripheral housing 12 and the quartz glass panel 9 in the box. The front, rear, left and right and the top quartz glass plates are tightly combined with the bottom stainless steel plate to form a sealed inner space. The top quartz glass plate has a small hole connected to the venting valve 11, and the bottom plate has a vent hole of the same size to ensure air circulation. The (quartz panel) inner wall panel 9 is made of quartz glass disposed in the outer casing 12, and the LED light source panel 10 is disposed between the casing 12 and the quartz inner wall panel 9. The three-dimensional light source is provided in the box from the five sides, and respectively The first side stereo LED illumination panel controller 1, the second side stereo LED illumination panel controller 2, the third side stereo LED illumination panel controller 3, the fourth side stereo LED illumination panel controller 4, the fifth side stereo LED The illumination panel controller 5 controls the full-wavelength LED device 10 to have a wavelength range of 200-1000 nm and a minimum wavelength adjustment of 5-10 nm. Also included is a heater 7a disposed at the bottom of the tank, a compressor 7b and connected to the UV lamp controller 6 at the top of the tank, and the UV lamp controller 6 displays the temperature and set temperature and automatically controls. The humidifier 8 is disposed in the cabinet and connected to the top UV lamp controller 6 of the cabinet to display humidity and set humidity, and is automatically controlled. Box plates 15a and 15b for space separation in the box. The culture bottle 13 in Fig. 2 is made of quartz glass. The bottle body 13a, the bottle cap 13b with a grinding mouth, the bottle mouth 13d, and the sterile vent plug insertion port 13c are all made of quartz glass to ensure the transparency of all wavelengths in the range of 200-1000 nm. The contact between the cap 13b and the mouth 13d is frosted to facilitate sealing.

Example 2:

A laboratory application method for a full-wavelength controllable light source biological culture device:

Biomaterial: Rhodiola callus

Medium: Optimized Rhodiola callus medium.

Photoreaction conditions study:

0-24 hours after inoculation of callus: dark culture, cell adaptation period.

2-30 days after inoculation: 620-630 nm light treatment, surface light intensity: 400 LUX (lux), the light wave condition for rapid cell division and growth, callus can grow rapidly. 30-60 days: 420-440 nm 50%+620-630 nm 50% (50% means that two light sources each form a mixed light source and simultaneously illuminate, the same reason), or 320-350 nm 40%+620- 630 nm 60%, or 280-320 nm 10%+620-630 nm 90% light treatment, surface light intensity: 400 LUX, optimized light wave conditions for different seedlings and rooting of Rhodiola. Can be repeatedly optimized to get the Rhodiola callus The best light wave combination for weaving growth and tissue culture seedling differentiation.

Example 3:

A full-wavelength controllable light source biological culture laboratory application method:

Biomaterials: Cordyceps sinensis i Cordyceps militaris t

Medium: Cordyceps militaris liquid medium

Photo-controlled fermentation program:

0-48 hours after inoculation: Dark culture to adapt and restore growth.

48-168 hours after inoculation: 620-630 nm, or 600-620 or 630-640 nm or 650-660 nm light treatment, surface light intensity: 400 LUX, repeated optimization to obtain the most hyphae growth and adenosine biosynthesis Good light wave conditions.

168-288 hours after inoculation: 380-400 nm or 400-410 nm or 410-420 nm or 420-440 nm light treatment, surface light intensity: 400 LUX, repeated optimization to obtain the best light wave for cordycepin and carotenoid biosynthesis condition.

Example 4:

Biomaterial: Ginseng isolated cells

Medium: Optimized ginseng cell liquid medium

Light control training program:

0-24 hours after inoculation: Culture in the dark to adapt and restore growth.

24-168 hours after inoculation: 620-630 nm light treatment, surface light intensity: 400 LUX, for optimal ginseng cell division to grow light wave conditions. At this time, the maximum ginseng cell biomass can be obtained.

168-288 hours after inoculation: 620-630 nm and 420-440 nm each at 50% ratio for light treatment, surface light intensity: 400 LUX, optimized ginsenoside biosynthesis light wave conditions. At this time, the largest ginsenoside can be obtained.

Claims

Claim

1. A biological culture device for a full-wavelength controllable light source, comprising a box body (12) and a culture bottle (13), wherein: in the box body (12):

First-sided stereo LED lighting panel controller (1), second-sided stereo LED lighting panel controller (2), third-sided stereo LED lighting panel controller (3), fourth-sided stereo LED lighting panel controller (4 ), the fifth-sided stereo LED lighting panel controller (5), the temperature, humidity and ultraviolet light controller (6) are respectively fixed on the inner side of the top panel of the cabinet, arranged from left to right;

The first stereo LED illumination panel controller (1) is connected to the second stereo LED illumination panel controller (2), the second stereo LED illumination panel controller (2) and the third stereo LED illumination panel controller ( 3) Connection, the third side stereo LED lighting panel controller (3) is connected with the fourth side stereo LED lighting panel controller (4), the fourth side stereo LED lighting panel controller (4) and the fifth side stereo LED lighting Panel controller (5) connection, fifth side stereo LED lighting panel controller (5) connected to temperature, humidity and UV lamp controller (6); digital display and parameter setting panel of each controller is exposed and located in the door The respective corresponding positions above;

The heater (7a), the compressor (7b), and the humidifier (8) are respectively fixed on the bottom plate of the casing;

The LED light source panel (10) is fixed on the inner wall of the corresponding cabinet (12); the inner wall panel (9) is covered on the LED light source panel (10); the other four inner walls in the cabinet are arranged in the same manner;

The first valve (11) is fixed above the top panel of the casing (12), and the vent hole of the first valve (11) penetrates the top of the casing (12) in a vertical portion to facilitate ventilation; the second valve (16) is located in the casing (12) a venting valve on the bottom plate, the second valve (16) also penetrates the bottom of the casing (12) in a vertical portion, and the first valve (11) and the second valve (16) are used for venting in the tank;

The first box door (14a) is fixed on the panel on the left side of the box body (12), and the second box door (14b) is fixed on the panel on the right side of the box body (12), and the two box doors are opposite; The plate (15a) is installed at a height of one-half of the height of the box body (12), and the second box plate (15b) is installed in a vertical center portion of the box body (12) in the left-right direction;

The culture bottle (13) includes a quartz bottle body (13a), a quartz bottle cap (13b), a sterile vent plug socket (13c), a quartz bottle body (13a) having a height of 5-20 cm, a diameter of 3-20 cm, and quartz. The inside of the bottle cap (13b) is ground and covered on the bottle mouth (13d). The outer surface of the bottle mouth (13d) is also ground to facilitate sealing with the quartz cap (13b). The sterile vent plug (13c) is located in the quartz. At the top of the cap (13b), the top of the quartz cap (13b) has an opening under the sterile vent plug (13c) to facilitate the insertion of sterile plugs to maintain sterile air into the bottle.

2. A biological culture apparatus for a full-wavelength controllable light source according to claim 1, wherein: said inner wall The panel (9) is a quartz panel.

3. A biological culture apparatus for a full-wavelength controllable light source according to claim 1, wherein: said LED light source panel (10) is provided with an LED lamp having a wavelength range of 200-1000 nm and an LED fluorescent lamp.