CN202139240U - Microalgae high-flux culture equipment - Google Patents
Microalgae high-flux culture equipment Download PDFInfo
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- CN202139240U CN202139240U CN 201120079934 CN201120079934U CN202139240U CN 202139240 U CN202139240 U CN 202139240U CN 201120079934 CN201120079934 CN 201120079934 CN 201120079934 U CN201120079934 U CN 201120079934U CN 202139240 U CN202139240 U CN 202139240U
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Abstract
The utility model provides equipment suitable for performing microalgae high-flux culture. The equipment comprises a frame, an oscillating mechanism, a temperature control device, a humidity control device, an airing control device, an object stage and a lighting device, wherein the object stage is suitable for fixing a culture structure used for containing algae strains or algae liquid; the lighting device is arranged below the object state, is suitable for providing uniform illumination for microalgae culture, and comprises a light source and a light guide structure; the light guide structure comprises a light guide layer; the side surface and the bottom surface of the light guide layer are provided with light reflection structures so as to guide light rays from the light source to mainly emit out from one surface of the light guide layer; therefore, the light rays from the light source is uniformly supplied to the culture structure contained with the algae strains or the algae liquid; the illumination uniformity and the illumination brightness used for the microalgae high-flux culture are improved; and the requirements on microalgae growth and high-flux screening are met.
Description
Technical field
The utility model relates to little algae commercial application field, and particularly, relates to being suitable in the high flux screening that is used in the algae kind and carry out the equipment that little algae high-throughput is cultivated.
Background technology
Little algae be one type on land, the ocean distributes extensively the high holophyte of nutritious, photosynthetic availability.Many little algaes can accumulate a large amount of greases in cell, fatty acid content can reach 80% of dried cell weight when some algae lacked at the nitrogen element, can from the little algae of this type, extract grease through the method for biorefinery, are used to prepare edible oil and biofuel; Little algae also can be used in the industrial processes such as wastewater treatment, to reduce environmental pollution.
The environmental friendliness characteristic of little algae, to the health care characteristic of humans and animals and the applications well property of bioenergy thereof; Caused global little algae bioenergy research and development upsurge; And gradually towards mass-producing, industrialization near; But commercial development also has many technical bottlenecks to need to break through, and the improvement of wherein little algae algae kind is the most important, and the screening of good little algae algae strain is the most important thing.The screening of good little algae algae strain must compare through they different performances, like the biochemical composition composition in the speed of growth, the cell, to the resistance of environment etc., and then takes all factors into consideration and selects the algae strain that is applicable to that mass-producing is cultivated.
(High throughput screening, HTS) technology is a kind of new and high technology that is used for aspects such as new compound exploitation and purpose strain improvement that grows up the eighties in 20th century to high flux screening.Its basic characteristics are to be carrier with the microwell plate, adopt the method for high-density, micro-robotization application of sample, can fast and parallel ground test great amount of samples.High-throughput techniques has the advantage of parallelization, robotization, traceization and intensification concurrently.High flux screening becomes drug development and microorganism field has a lot of comparatively successful application.
Utilizing little algae to prepare in the research of biofuel and high value added product, the screening of algae kind occupies an important position, and wherein high flux screening will play an important role, and reduces in the research process consumption for human and material resources and time widely.In addition, can also strong technical guarantee be provided, advance the process of little algae related industriesization for the bigger genetic engineering breeding of development potentiality.Rational and effective cultural method and condition control are cultivated for algae kind high-throughput has important effect.
At present; The system that the laboratory is used for the cultivation of algae kind high-throughput is relatively simple; The cultivation of algae strain generally all is to place growth cabinet in the triangular flask system, and stuffiness is cultivated or simply inserted the ventpipe aerobic culture, and illumination and temperature are regulated and control by growth cabinet; Temperature control is more even, but this mode is difficult to realize the consistent of illumination and ventilatory effect.Also have a kind of culture systems to place shaking table exactly in addition; Illumination and temperature are regulated and control by shaking table; Through shaking the accelerating gas exchange; The ventilatory effect of this mode is consistent relatively with temperature, but faces the problem that uneven illumination is even, intensity of illumination can't satisfy algal grown and high flux screening needs equally.In addition, also there is the big and micro algae growth of system problem slowly in above-mentioned dual mode.
And existing illumination box or illumination shaking table are to adopt fluorescent tube to carry out illumination, not only exist outside the problems such as uneven illumination is even, the illuminance decay is obvious, also have defectives such as heat production is big, the life-span short, the illuminance adjusting is discontinuous.Such culture apparatus only is fit to adopt the triangular flask training method at present, if be used to cultivate microminiaturized laboratory sample, these problems can be more outstanding.In addition, the top light of traditional shaking table photograph and vessel fixed form have also limited the use of microwell plate, aspect solution ventilation, mixing, crossed contamination, evaporation and illumination, can't satisfy microminiaturized high-throughout experimental technique and implement especially.
The utility model content
In view of this, the purpose of the utility model is to solve the problems referred to above of existing in the prior art and at least one aspect of defective.
One of purpose of the utility model is to provide a kind of equipment that little algae high-throughput is cultivated that is suitable for carrying out, and it can improve illumination uniformity coefficient and the intensity of illumination that is used for little algae high-throughput cultivation, satisfies the needs of micro algae growth and high flux screening.
An aspect according to the utility model; A kind of equipment that little algae high-throughput is cultivated that is suitable for carrying out is provided; Comprise framework, oscillating mechanism, temperature-control device, humidity conditioner, ventilation controller, Stage microscope and means of illumination, wherein, Stage microscope is suitable for being fixed for holding the cultivation structure of algae kind or algae liquid; Means of illumination is arranged on the below of Stage microscope; Be suitable for little algae and cultivate uniform illumination is provided, and means of illumination comprises light source and light guide structure, and light guide structure is configured to be suitable for the light from light source is provided to the cultivation structure that accommodates algae kind or algae liquid equably.
Be suitable for carrying out in the equipment of little algae high-throughput cultivation above-mentioned; This light guide structure can be multilayered structure; Comprise: optical waveguide layer, be suitable for the light from light source is reflected to all directions equably, incide the optical waveguide layer from a pair of side of optical waveguide layer from the light of light source; The bottom reflection film, the bottom surface of covering optical waveguide layer is suitable for returning optical waveguide layer by optical waveguide layer to the luminous reflectance of said bottom reflection; With the offside reflection film, another of covering optical waveguide layer is suitable for returning optical waveguide layer by optical waveguide layer to said another luminous reflectance to offside reflection the side.
Be suitable for carrying out the equipment that little algae high-throughput is cultivated above-mentioned, the ratio of each the lateral area in the area of the end face of optical waveguide layer and the said a pair of side can be greater than 10: 1.
Be suitable for carrying out on the submarginal summit portion of optical waveguide layer, can also being provided with reflectance coating in the equipment of little algae high-throughput cultivation above-mentioned, last reflectance coating is suitable for the light reflected back optical waveguide layer from end face periphery inclination outgoing.
Carry out in the equipment of little algae high-throughput cultivation above-mentioned being suitable for, at least one in said bottom reflection film and the offside reflection film can be total reflection film.
Be suitable for carrying out in the equipment of little algae high-throughput cultivation above-mentioned; Optical waveguide layer can be made up of base material and the leaded light powder or the leaded light particle that are evenly distributed in the base material; The light that gets into optical waveguide layer reflects when meeting with leaded light powder or leaded light particle, at least a in reflection and the transmission; And base material can comprise polymethyl methacrylate materials, and leaded light powder or leaded light particle can comprise nanostructure.
Be suitable for carrying out in the equipment that little algae high-throughput cultivates above-mentioned, this multilayered structure can also comprise all light film that is positioned on the optical waveguide layer end face, is suitable for the upwards emission equably of light from the end face outgoing of optical waveguide layer.
Be suitable for carrying out in the equipment that little algae high-throughput cultivates above-mentioned, this multilayered structure can also comprise equal tabula rasa, is arranged on the top of optical waveguide layer, is suitable for the light from the end face outgoing of optical waveguide layer upwards is transmitted into the cultivation structure that accommodates algae kind or algae liquid equably.
Be suitable for carrying out going up equal tabula rasa and all can separating predetermined distance between the light film in the equipment that little algae high-throughput cultivates above-mentioned, with at last equal tabula rasa with all form the space of being scheduled between the light film.
Be suitable for carrying out in the equipment that little algae high-throughput cultivates above-mentioned, at last equal tabula rasa with all can be provided with bracing or strutting arrangement between the light film, this bracing or strutting arrangement is suitable for adjusting the said predetermined distance that separates, to adjust said predetermined spatial volume.
Carry out in the equipment of little algae high-throughput cultivation above-mentioned being suitable for, said light source can comprise led array, and between led array and light guide structure, can be provided with lens, and these lens are suitable for projecting in the optical waveguide layer behind the optical convergence from led array.
Above-mentionedly be suitable for carrying out the equipment that little algae high-throughput cultivates and can also comprise light intensity controller, it provides the light intensity to the current control of led array from the led array outgoing through control.
Be suitable for carrying out in the equipment of little algae high-throughput cultivation above-mentioned; Framework is suitable for forming and carries out little algae cultured microalgae culture environment; Wherein Stage microscope and means of illumination are arranged in this framework; Temperature-control device is suitable for controlling the temperature in little algae culture environment, and humidity conditioner is suitable for controlling the humidity in little algae culture environment, and ventilation controller is suitable for controlling the ventilation in little algae culture environment; And oscillating mechanism is suitable for vibrating in little algae culturing process, and wherein means of illumination is arranged on the oscillating mechanism.
Description of drawings
Consult follow-up diagram and describe above-mentioned and further feature and the advantage that to understand the utility model better.Not detailed row and non-restrictive example then please refer to the description of this subsequent figures in the literary composition.Element in the diagram might not meet ratio, and is the principle of depicting the utility model with the mode of stressing.In each diagram, components identical is represented with identical Reference numeral in the difference diagram.In the accompanying drawings:
Fig. 1 is the synoptic diagram of component part that is suitable for carrying out the equipment that little algae high-throughput cultivates that illustrates according to the utility model;
Fig. 2 shows the shaker platform of means of illumination that is suitable for carrying out the equipment that little algae high-throughput cultivates that wherein is provided with according to the utility model;
Fig. 3 A is the enlarged diagram of the A part among Fig. 2;
Fig. 3 B is the enlarged diagram of the B part among Fig. 2;
Fig. 3 C is the enlarged diagram of the C part among Fig. 2;
Fig. 3 D is the enlarged diagram of the D part among Fig. 2;
Fig. 4 is the synoptic diagram that illustrates from the direct of travel of light in light guide structure of light source; And
Fig. 5 illustrates the microminiaturized cultivation of frustule of the high-throughput culture device of utilizing the utility model and the dependency under the conventional training method.
Embodiment
To combine accompanying drawing and embodiment that the utility model is done further explanation below.
Fig. 1 shows the equipment that is suitable for carrying out little algae high-throughput cultivation according to the utility model.As shown in Figure 1, be suitable for carrying out the equipment 100 that little algae high-throughput cultivates and comprise casing or skeleton construction 1.Body structure 1 is divided into two portions, in the left part in Fig. 1, is provided with shaker platform 2 and the transmission rig 3 that supports shaker platform 2 and be suitable for shaker platform 2 is vibrated, and can realize the shaking culture of little algae thus.Wherein, shaker platform 2 is provided with means of illumination 5, and it is used for having microwell plate of little algae or porous plate 10 that illumination is provided to the inoculation that is placed on shaker platform 2 tops.Porous plate 10 can comprise 96 orifice plates, 48 orifice plates, 24 orifice plates and 12 orifice plates etc.According to a kind of embodiment of the utility model, porous plate 10 can be cultivated bearing structure by other little algae and replace, as shakes bottle or triangular flask etc.
In one embodiment; Porous plate 10 is fixed in the body structure 1 through Stage microscope (not shown); Means of illumination 5 is positioned at the below of porous plate 10; Supply with light source from its bottom, thereby be convenient to, can prevent crossed contamination, guarantee ventilation, evaporate consistence, increase rotating speed, increase ventilatory effect etc. the porous plate top seal.
In body structure 1; Can also be provided with TP 6, add wet hole 7, heating blower 8, refrigeration fan 9, vaporizer and compressor (not shown) etc.; Be provided with gear 4 in the left part in Fig. 1; Thus to the environment of culturing microalgae in the body structure 1, comprise that temperature, humidity, ventilation etc. control, realize being suitable for the good environment that little algae is cultivated.
Fig. 2 A shows the shaker platform that is suitable for carrying out the means of illumination in the equipment that little algae high-throughput cultivates that wherein is provided with according to the utility model.Wherein in Fig. 2, the lower left is the vertical view of shaker platform, and the upper left side is the sectional view of edge I-I line intercepting wherein, and the lower right is the sectional view of edge II-II line intercepting wherein.According to a kind of embodiment of the utility model, means of illumination 5 is arranged in the shaker platform 2, can shake with shaker platform 2, and have the porous plate 10 of little algae that illumination is provided to the inoculation of shaker platform 2 tops, can carry out wave and culture to little algae thus.
As shown in Figure 2, shaker platform 2 surfaces are provided with a plurality of perforates 201, like square opening, make and can pass perforate 201 projections to porous plate from the light that is arranged on shaker platform 2 subsurface means of illumination 5.Can fit together through screw 202 etc. between shaker platform 2 and the means of illumination under it 5.According to the embodiment of the utility model, means of illumination 5 is between upper cover plate 203 and base plate 204.Upper cover plate 203 only covers the bight part of means of illumination 5, so that beam projecting.Base plate 204 covers means of illumination 5 fully and provides support to it.
As shown in Figure 2, means of illumination 5 comprises light source 501 and light guide structure, and it is arranged on the both sides of light guide structure, preferably is arranged on the relative both sides of light guide structure, incident beam in light guide structure.According to a kind of embodiment of the utility model, light source 501 adopts led array, this can overcome, and heat radiation such as conventional light sources such as fluorescent tubes is big, uneven illumination even, the illuminance decay obviously, short, illuminance of life-span regulates defectives such as discontinuous.Be supplied to the electric current of led array through adjusting, can regulate its intensity of illumination.
In the utility model, light guide structure is configured to be suitable for the light from light source the porous plate 10 that little algae is arranged to inoculation being provided equably.The embodiment of the utility model adopts the bottom to supply the light mode, and means of illumination 5 is arranged on the below of porous plate 10, and the light guide structure guiding is upwards throwed towards porous plate 10 from the light of light source 501 equably.
Fig. 3 A-3D shows in detail the structure according to the means of illumination of the utility model; Wherein Fig. 3 A is the enlarged diagram of the A part among Fig. 2; Fig. 3 B is the enlarged diagram of the B part among Fig. 2; Fig. 3 C is the enlarged diagram of the C part among Fig. 2, and Fig. 3 D is the enlarged diagram of the D part among Fig. 2.
Embodiment according to the utility model; Shown in Fig. 3 A and 3B; Light guide structure is a multilayered structure; Comprise optical waveguide layer or light guiding plate 503 and the bottom reflection film 504 that covers optical waveguide layer 503 bottom surfaces, optical waveguide layer 503 be suitable for from light source 501 incident light equably to all directions reflection, refraction or transmission, bottom reflection film 504 is suitable in the light reflected back optical waveguide layer 503 with directive optical waveguide layer 503 bottom surfaces.Below bottom reflection film 504, reflector 506 can also be set, its can be further with in the light of directive optical waveguide layer 503 bottom surfaces and the light reflected back optical waveguide layer 503 that penetrates bottom reflection film 503.
According to a kind of embodiment of the utility model, on the end face of optical waveguide layer 503, can also be provided with all light films 507, all light film 507 can carry out homogenizing to the light from optical waveguide layer 503 outgoing.In the another kind of embodiment of the utility model, on the submarginal summit portion of optical waveguide layer 503, reflectance coating 509 can also be set, last reflectance coating 509 is suitable for attempting from the light reflected back optical waveguide layer 503 of end face periphery inclination outgoing.
In the utility model; Reflector 506, bottom reflection film 504, optical waveguide layer 503, all light film 507, go up reflectance coating 509 and be arranged in order between base plate 204 and the upper cover plate 203, and be formed with in the upper cover plate 203 and be suitable for the opening that passes through from the light of optical waveguide layer 503 outgoing.According to a kind of embodiment of the utility model, can also be provided with all tabula rasas 508 above the optical waveguide layer 503, it is positioned at the following of upper cover plate 203 and supports upper cover plate 203, and is suitable for the light of further homogenizing from optical waveguide layer 503 outgoing.
According to a kind of embodiment of the utility model, shown in Fig. 3 A-3C, going up can be spaced a predetermined distance between equal tabula rasa 508 and the optical waveguide layer 503, so that between last equal tabula rasa 508 and optical waveguide layer 503, form certain space 510.From the light of optical waveguide layer 503 outgoing in this space 510 with the air collision of for example air and so on, diffuse-reflectance takes place, make that the light of outgoing is more even.In one embodiment, between last equal tabula rasa 508 and optical waveguide layer 503, be provided with adjustable for height bracing or strutting arrangement, the support copper post 511 shown in Fig. 3 B; It can adjust the size of above-mentioned predetermined distance; So that adjust the volume in said space 510, irreflexive degree takes place thereby can adjust from the light of optical waveguide layer 503 outgoing, promptly said space is big more; It is abundant more that diffuse-reflectance is carried out, and the light of outgoing is even more.
According to the embodiment of the utility model, shown in Fig. 3 A and 3B, led array 501 is arranged on the substrate 502, like aluminium base.Substrate 502 is fixed on the side group plate 205, and led array 501 is to two light incident side face throw lights of optical waveguide layer 503.Side group plate 205 can be L shaped shape, covers the side of light guide structure fully, and below light guide structure, extends certain-length.The side group plate 205 that extends below the light guide structure is fixing through the coupling device such as screw 206 with base plate 204, to form skeleton construction.Between optical waveguide layer 503 and the side group plate 205 and on all between tabula rasa 508 and the optical waveguide layer 503 bracing or strutting arrangement 208 and 209 such as packing ring can be set.
In the utility model, shown in Fig. 3 C, two other place, side at light guide structure promptly in non-two sides (for example, perpendicular to two sides of light incident side face) that receive light of optical waveguide layer 503, also is provided with offside reflection film 505.Light from led array 501 wherein incides in the optical waveguide layer 503 after lens 511 (shown in Fig. 3 D) are assembled; Through bottom reflection film 504 and the end face outgoing of offside reflection film 505 reflection backs from optical waveguide layer 503, projecting inoculation equably has on the porous plate 10 of little algae.
Fig. 3 D shows the detailed structure of light source.Wherein, led array 501 is electrically connected to substrate 502, on each LED is unitary, is provided with lens 511, and lens 511 are used for the optical convergence that penetrates from led array to optical waveguide layer 503.
Fig. 4 illustrates light path or the direct of travel of incident light in light guide structure from led array in more detail.As shown in Figure 4, optical waveguide layer 503 is made up of base material 5030 and the leaded light powder or the leaded light particle 5031 that are evenly distributed in the base material 5030.Base material 5030 for example can be polymethylmethacrylate (PMMA) material, and it has high refractive index usually.Leaded light powder or leaded light particle 5031 can be nanometer leaded light powder or particle.
Embodiment according to the utility model; The light beam that led array 501 sends projection after lens 511 are assembled gets in the optical waveguide layer 503; Because leaded light particle 5031 distributes in base material 5030 equably; Therefore every bundle light get into must have behind the optical waveguide layer two kinds maybe, the one, move ahead along original route, another kind then be meet with leaded light particle 5031 reflect, reflection or transmission.The light that gets into optical waveguide layer 503 experienced repeatedly reflect, reflect after, six of the directive optical waveguide layer faces equably.Owing to receive two sides of light to locate to be provided with respectively reflectance coating or reflector with the bottom surface optical waveguide layer non-; As 504,506 and 505; The light that the bottom reflection film 504 that covers optical waveguide layer 503 bottom surfaces will be attempted to penetrate from optical waveguide layer 503 bottom surfaces is reflected back optical waveguide layer 503 once more; Offside reflection film 505 will be attempted to receive light that two sides of light penetrate reflected back optical waveguide layer 503 once more from optical waveguide layer 503 non-, and final light can only be from the end face and the outgoing of two light incident side faces of optical waveguide layer 503.According to the utility model, the area of the end face of optical waveguide layer 503 is set to much larger than the area of light incident side face, and then all light is almost all from the top surface side outgoing equably of optical waveguide layer 503.A kind of embodiment according to the utility model; The end face of optical waveguide layer is set to greater than about 10: 1 with said another ratio to each the lateral area in the side; Making almost all has the porous plate 10 of little algae from the light of light source entering light guide structure from the even outgoing of top surface side to the inoculation of optical waveguide layer, thereby improves light efficiency.
From the light of the top surface side outgoing of optical waveguide layer 505 through equal light films 507 with on all tabula rasa 508 and project behind the gas homogenization the space therebetween above inoculation the porous plate 10 of little algae is arranged, little algae algae kind is carried out illumination cultivation.According to a kind of embodiment of the utility model, for fear of the loss of incident beam in light guide structure, bottom reflection film 504 can be made up of total reflection layer or total reflection film with offside reflection film 505.Below will come verification setting that the performance of the shaking table that is used for the means of illumination that little algae high-throughput cultivates of the utility model is arranged through specific embodiment.
Whether the light distribution of in the present embodiment, estimating the shaking table that is designed through the light intensity of measuring the different positions place reaches the requirement of little algae high flux screening.
Experimental procedure does, the temperature of setting shaking table is 25 ℃, and light intensity is 4000lux, measures the intensity of illumination of the shaking table upper left corner, the upper right corner, the lower left corner, the lower right corner and the center position that are designed with luxmeter.During mensuration with 96 microwell plates as measuring basis.
Data processing adopts SPSS Statistics 17.0 softwares.Through to the illumination homogeneity between the calculating of the variation coefficient (CV) of place, each hole in each microwell plate intensity of illumination and each hole that one-way analysis of variance (ANOVA) is estimated each microwell plate.Through the illumination homogeneity that the calculating and the one-way analysis of variance (ANOVA) of the variation coefficient (CV) of light intensity MV in 5 microwell plates of different positions are estimated the different positions place.In the above data processing with the measurement of error P＜0.05 as level of signification.
CV through calculating between each hole of each microwell plate is respectively: the upper left corner 9.32%, the upper right corner 9.85%, the lower left corner 9.43%, the lower right corner 10.22%, center 8.34%.Obtain the degree of freedom F＞F between each hole of each microwell plate through one-way analysis of variance
0.05Therefore, error P＜0.05 explains that the growth differences of the algae strain between each hole does not reach level of signification, explains that promptly light distribution between each hole of microwell plate of position than homogeneous, can satisfy the high flux screening of little algae.
CV through calculating between the microwell plate of position is 9.58%, and variance analysis obtains F＞F
0.05, therefore P＜0.05 explains that the difference between light intensity does not reach level of signification between each microwell plate, the light distribution at promptly whole Stage microscope different positions place is a homogeneous, can satisfy the high flux screening of little algae.
In the present embodiment; Select 96 orifice plates, 48 orifice plates, 24 orifice plates, 12 orifice plates respectively; Adopt the means of illumination in the utility model that illumination is provided; Algae strain Chlorella vulgaris (purchase in hydrobiont institute of Chinese Academy of Sciences algae kind storehouse, be numbered 1068) is cultivated, adopt BG11 substratum (1-3 sees the following form).
The composition and the content thereof of table 1 BG11 substratum
| Sequence number | Composition | Amount (/L) | |
| 1 | NaNO 3 | 1.5 |
|
| 2 | MgSO 4·7H 2O | 0.075 |
|
| 3 | CaCl 2·2H 2O | 0.036 |
|
| 4 | Citric?acid | 0.006g | |
| 5 | The Fe-EDTA | 1mL | |
| 6 | Na 2CO 3 | 0.02g | |
| 7 | A 5?Trace?element | 1.0ml |
Table 2 A
5Trace element forms and content
| Sequence number | Composition | Amount (g/L) |
| 1 | H 3BO 3 | 2.86 |
| 2 | MnCl 2·4H 2O | 1.81 |
| 3 | ZnSO 4·7H 2O | 0.222 |
| 4 | Na 2MoO 4·2H 2O | 0.39 |
| 5 | CuSO 4·5H 2O | 0.079 |
| 6 | Co(NO 3) 2·6H 2O | 0.0494 |
The composition and the content thereof of table 3 Fe-EDTA liquid storage
| Sequence number | Composition | Amount (g/L) |
| 1 | FeCl 3·6H 2O | 3.15 |
| 2 | Na 2EDTA·2H 2O | 4.36 |
The algae strain of exponential phase of growth of some amount is inoculated in the sterilized substratum of 300mL by 20% inoculum size; Mix the back with 8 passage pipettors, measure corresponding algae liquid by following liquid feeding and join in each microwell plate: the liquid amount of 96 microwell plates, 48 microwell plates, 24 microwell plates, 12 microwell plates is respectively 200 μ L, 500 μ L, 1000 μ l, 2000 μ l.Every type plate do 3 parallel, cover plate lid sealing (above experiment is all carried out) at last under aseptic condition.Other experiment condition is: 25 ℃ of temperature, illumination 30 μ molm
-2S
-1, photoperiod 12hL/12hD, shaking speed 180rpm.Cultivate and measure the OD in each hole respectively after 14 days
750(expression algae cell density).
Analyze the algae strain living weight in each hole of same type microwell plate, obtain that the CV data see the following form between the hole of each orifice plate.Data are visible from following table 4; No matter adopt the microwell plate (96 orifice plates, 48 orifice plates, 24 orifice plates, 12 orifice plates) of which kind of type; The collimation of frustule living weight is all very good between each hole, this show the means of illumination that is provided with in the utility model the high-throughput culture device be applicable to various types of microwell plates on the market.
Table 4
| The orifice plate type | CV between the hole | The crossed |
| 96 holes | 10.2％ | 0 |
| 48 holes | 7.6％ | 0 |
| 24 holes | 6.4％ | 0 |
| 12 holes | 6.3％ | 0 |
In an embodiment, will to the frustule of the high-throughput culture device of utilizing the utility model microminiaturized cultivate and conventional training method under dependency analyze.Through three kinds of different algae kinds under certain test conditions, the energy for growth in shaking bottle and microwell plate relatively estimate with the high-throughput culture device of the utility model to come culturing microalgae through microwell plate and whether have higher dependency with shake-flask culture between little algae.
Testing used algae kind is respectively: Chlorella vulgaris; Scenedesmus quadricauda; Oscillatoria sp (above-mentioned algae kind is all purchased in hydrobiont institute of Chinese Academy of Sciences algae kind storehouse); All adopt BG11 substratum (seeing table 1), the microwell plate type of selecting for use is 48 holes, through silica-gel plate lid sealed micro orifice plate.
Experimental procedure is: the algae liquid that some amount is in logarithmic phase is inoculated in the sterilized substratum by 20% inoculum size; The liquid amount that shakes bottle is 100mL/250mL; The liquid amount of 48 microwell plates is 500 μ L; Shake bottle and seal with 8 layers of gauze, microwell plate is with silica-gel plate lid sealing (above experiment is all carried out under aseptic condition).The culture condition that experiment is set is: 25 ℃ of temperature, illumination 30 μ molm
-2S
-1, photoperiod 12hL/12hD, shaking speed 180rpm, culture cycle is 14 days.Regularly get the algae liquid of 2mL every day in the cultivation from shake bottle, and the algae liquid of from 48 microwell plates, getting 5 holes carries out OD
750The mensuration of (expression algae cell density).Be X-coordinate at last with time, OD
750For ordinate zou is drawn out the growth curve of each algae kind, as shown in Figure 5, again to shaking the OD at each time point place in bottle and the microwell plate
750Carrying out curve estimates and linear regression processing.Wherein, in Fig. 5, Cv, Sq, Os represent algae kind Chlorella vulgaris respectively, Scenedesmus quadricauda, and Oscillatoria sp, MTP represent the growth curve in the microwell plate.
Can obtain each algae kind from Fig. 5 through above-mentioned data processing and all show characteristics separately, explain that used algae kind high flux screening device can be not influential to the characteristic of each algae kind self in the variation tendency of shaking growth curve on bottle and the microwell plate.And through to shaking the OD at each time point place in bottle and the microwell plate
750Linear regression processing can obtain the dependency R value of each algae kind under two kinds of training methods and be respectively R
Cv=0.986, R
Sq=0.991, R
Os=0.985, promptly these two kinds of training methods show higher dependency in used algae kind high flux screening device.
Can find out according to above description; In the utility model; Through offside reflection film and bottom reflection film at optical waveguide layer; So that guiding is mainly penetrated from the end face of optical waveguide layer from the light of light source, improve the illumination uniformity coefficient and the intensity of illumination that are used for little algae high-throughput cultivation, satisfy the needs of micro algae growth and high flux screening.And; Adopt led array as light source; Can overcome that heat radiation such as conventional light sources such as fluorescent tubes is big, uneven illumination even, the illuminance decay obviously, short, illuminance of life-span regulates defectives such as discontinuous; Be supplied to the electric current of led array through adjusting, can easily regulate its intensity of illumination, thereby realize that the algae kind cultivates under the homogeneous illumination condition.In addition, have through adjusting different optical bands the LED lamp quantity with arrange, adjust the illumination condition that is fit to micro algae growth.According to the means of illumination of the utility model, can be set to uniform twinkler by whole shaking table table top, can satisfy the use of various forms microwell plate or porous plate simultaneously, do not receive the restriction of orifice plate form structure.And the utility model adopts the bottom to supply the light mode, is convenient to the top seal with porous plate, can prevent crossed contamination, assurance ventilation, evaporation consistence, increases rotating speed, increase ventilatory effect etc.
At present, the device that does not have little algae high-throughput to cultivate as yet, those skilled in the art cultivate for the high-throughput of realizing little algae, once attempt light source is installed in the microbiological incubator; For not influencing experimental implementation, can only be installed in the top or the side of microbiological incubator, still; Top or side confession photoconduction cause incubator inside and receive light inhomogeneous, influence The selection result, and light path are longer; The illuminance decay is high, and at the bottom of the light utilization efficiency, illumination is regulated insensitive.The trial that designs kinds of schemes through the utility model people is implemented in Stage microscope bottom confession light, has realized that illumination is even in little algae high-throughput cultivation, reduces the illuminance decay, improves light utilization efficiency, and illumination adjusting sensitivity can satisfy the demand that little algae high-throughput is cultivated.The led array that the means of illumination of the utility model adopts does not have the many shortcomings of common fluorescent tube heat radiation.
Means of illumination according to the utility model can combine by conventional shaking table; Can realize little algae shake under the state control illumination, temperature control, control is wet and the high-throughput of control gas is cultivated; In suitable algae plant height flux culture condition scope, temperature, humidity, speed, illuminance are adjustable continuously.And the high-throughput culture device of the utility model can adopt the module stacked system, stackable many groups, and every pack module can independent temperature control, control is wet, control speed, control light, further improves the handiness that it is used to screen comparative study.Further, the means of illumination in the utility model adopts means such as sidelight irradiation, diffuse-reflectance, even light can realize illuminance more uniformly.Thus, be suitable for carrying out the breeding techniques such as screening, selection by mutation and genetic engineering breeding that equipment that little algae high-throughput cultivates can be applied to a large amount of algae kinds of nature according to the utility model.
Though below described the utility model, it will be recognized by those skilled in the art, under the prerequisite of marrow that does not depart from the utility model or scope, can change in form and details with reference to preferred embodiment.
Claims (12)
1. one kind is suitable for carrying out the equipment that little algae high-throughput is cultivated, and comprises framework, oscillating mechanism, temperature-control device, humidity conditioner, ventilation controller, Stage microscope and means of illumination, it is characterized in that,
Stage microscope is suitable for being fixed for holding the cultivation structure of algae kind or algae liquid,
Means of illumination is arranged on the below of Stage microscope, and be suitable for little algae cultivation uniform illumination is provided, and
Means of illumination comprises light source and light guide structure, and light guide structure is configured to be suitable for the light from light source is provided to the cultivation structure that accommodates algae kind or algae liquid equably.
2. according to claim 1 being suitable for carried out the equipment that little algae high-throughput is cultivated, and it is characterized in that this light guide structure is a multilayered structure, comprising:
Optical waveguide layer is suitable for the light from light source is reflected to all directions equably, incides the optical waveguide layer from a pair of side of optical waveguide layer from the light of light source;
The bottom reflection film, the bottom surface of covering optical waveguide layer is suitable for returning optical waveguide layer by optical waveguide layer to the luminous reflectance of said bottom reflection; With
The offside reflection film, another of covering optical waveguide layer is suitable for returning optical waveguide layer by optical waveguide layer to said another luminous reflectance to offside reflection the side.
3. according to claim 2 being suitable for carried out the equipment that little algae high-throughput is cultivated, it is characterized in that,
The ratio of each the lateral area in the area of the end face of optical waveguide layer and the said a pair of side was greater than 10: 1.
4. the equipment that is suitable for carrying out little algae high-throughput cultivation according to claim 2; It is characterized in that; On the submarginal summit portion of optical waveguide layer, also be provided with reflectance coating, last reflectance coating is suitable for the light reflected back optical waveguide layer from end face periphery inclination outgoing.
5. according to claim 2 being suitable for carried out the equipment that little algae high-throughput is cultivated, it is characterized in that,
In said bottom reflection film and the offside reflection film at least one is total reflection film.
6. according to claim 2 being suitable for carried out the equipment that little algae high-throughput is cultivated, and it is characterized in that this multilayered structure also comprises:
Be positioned at all light film on the optical waveguide layer end face, be suitable for upwards emission equably of light from the end face outgoing of optical waveguide layer.
7. according to claim 6 being suitable for carried out the equipment that little algae high-throughput is cultivated, and it is characterized in that this multilayered structure also comprises:
Go up equal tabula rasa, be arranged on the top of optical waveguide layer, be suitable for the light from the end face outgoing of optical waveguide layer upwards is transmitted into the cultivation structure that accommodates algae kind or algae liquid equably.
8. be suitable for carrying out the equipment that little algae high flux is cultivated based on claim 7 is described, it is characterized in that, go up equal tabula rasa and all separate predetermined distance between the light film, with at last equal tabula rasa with all form the space of being scheduled between the light film.
9. the equipment that is suitable for carrying out little algae high-throughput cultivation according to claim 8; It is characterized in that; At last equal tabula rasa with all be provided with bracing or strutting arrangement between the light film, this bracing or strutting arrangement is suitable for adjusting the said predetermined distance that separates, to adjust said predetermined spatial volume.
10. according to claim 1 and 2 being suitable for carried out the equipment that little algae high-throughput is cultivated, it is characterized in that,
Said light source comprises led array, and
Between led array and light guide structure, be provided with lens, these lens are suitable for projecting in the optical waveguide layer behind the optical convergence from led array.
11. according to claim 10 being suitable for carried out the equipment that little algae high-throughput is cultivated, and it is characterized in that this equipment also comprises:
Light intensity controller provides the light intensity to the current control of led array from the led array outgoing through control.
12. according to claim 1 and 2 being suitable for carried out the equipment that little algae high-throughput is cultivated, it is characterized in that,
Framework is suitable for forming and carries out little algae cultured microalgae culture environment, and wherein Stage microscope and means of illumination are arranged in this framework,
Temperature-control device is suitable for controlling the temperature in little algae culture environment,
Humidity conditioner is suitable for controlling the humidity in little algae culture environment,
Ventilation controller is suitable for controlling the ventilation in little algae culture environment, and
Oscillating mechanism is suitable for vibrating in little algae culturing process, and wherein means of illumination is arranged on the oscillating mechanism.
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| CN 201120079934 CN202139240U (en) | 2011-03-15 | 2011-03-15 | Microalgae high-flux culture equipment |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102212471A (en) * | 2011-03-15 | 2011-10-12 | 新奥科技发展有限公司 | Microalgae high-throughout culture equipment |
| CN107760586A (en) * | 2017-09-21 | 2018-03-06 | 浙江海洋大学 | A kind of microalgae biofilm system of Immobilized culture |
-
2011
- 2011-03-15 CN CN 201120079934 patent/CN202139240U/en not_active Expired - Lifetime
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102212471A (en) * | 2011-03-15 | 2011-10-12 | 新奥科技发展有限公司 | Microalgae high-throughout culture equipment |
| WO2012126328A1 (en) * | 2011-03-15 | 2012-09-27 | 新奥科技发展有限公司 | Lighting device, high throughput cultivation equipment comprising same |
| CN102212471B (en) * | 2011-03-15 | 2013-08-21 | 新奥科技发展有限公司 | Microalgae high-throughout culture equipment |
| CN107760586A (en) * | 2017-09-21 | 2018-03-06 | 浙江海洋大学 | A kind of microalgae biofilm system of Immobilized culture |
| CN107760586B (en) * | 2017-09-21 | 2021-01-05 | 浙江海洋大学 | Immobilized culture microalgae biofilm system |
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