CN106497792B - A kind of cultural method of photo-biological cell - Google Patents
A kind of cultural method of photo-biological cell Download PDFInfo
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- 229940082787 spirulina Drugs 0.000 description 4
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- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 description 2
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- 235000002470 Asclepias syriaca Nutrition 0.000 description 1
- 244000000594 Asclepias syriaca Species 0.000 description 1
- 241000195649 Chlorella <Chlorellales> Species 0.000 description 1
- 241000180279 Chlorococcum Species 0.000 description 1
- 241000192700 Cyanobacteria Species 0.000 description 1
- 241000168525 Haematococcus Species 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
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- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
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Abstract
The embodiment of the invention provides a kind of cultural methods of photo-biological cell, are related to the culture field of photo-biological cell, can convenient, effectively eliminate the bubble in the attachment culture of photo-biological cell, further increase its reproductive efficiency.This method includes that step (1) judges that bubble whether there is between carrier layer and cloth liquid layer, or is present between carrier layer and cellular layer；Step (2), the position occurred according to bubble, adjust the default supply amount of culture solution, until bubble is fully drained；Including when bubble is present between carrier layer and cloth liquid layer, increasing the supply amount of culture solution, until bubble is fully drained；When bubble is present between carrier layer and cellular layer, stop, or stop the supply of culture solution repeatedly, until bubble is fully drained；Step (3), the default supply amount according to culture solution continue to cultivate photo-biological cell.Culture for photo-biological cell.
Description
Technical field
The present invention relates to the culture field of photo-biological cell more particularly to a kind of cultural methods of photo-biological cell.
Background technique
Extensive, the high yielding culture of photo-biological cell (such as microalgae) are the bottleneck for restricting its industrialized development, low cost, height
The exploitation for imitating training method is the key that large-scale production.Existing training method is specially being opened based on " water body culture "
It puts in formula raceway pond, closing or semi-enclosed reactor and mixes photo-biological cell and nutrients, water, and provide it suitable
Suitable illumination makes its cell division breeding nature growth.
But it finds in practical applications, when being cultivated using water body training method photo-biological cell, light utilization ratio
Room for promotion it is limited, so that above-mentioned training method is not able to satisfy the technical need of current photo-biological cell large-scale culture.
In order to preferably improve the light utilization efficiency of photo-biological cell, the mode of " attachment culture " is applied to photo-biological cell
Culture.As shown in Figure 1, attachment culture is specifically to be arranged one layer in support layer surface to have the cloth liquid layer for holding liquid ability, by light
Biological cell is fixed or be attached on the culture carrier (carrier layer i.e. in figure) on cloth liquid layer surface or surface and inside, leads to
It crosses to cell sustainable supply culture solution and cell is made to grow to form cellular layer in the carrying layer surface schizogamy on cloth liquid layer surface.
However, being found during using above method culture photo-biological cell, in the case where illumination is good, temperature is high
(such as noon), the carbon dioxide gas cognition dissolved in culture solution release, and photo-biological cell metabolism can also generate oxygen, therefore
Cause between carrier layer and cellular layer, between cloth liquid layer and carrier layer, easily formation bubble.Bubble will lead to cellular layer and can not obtain
Supplied from cloth liquid layer and the culture solution of carrier layer, in turn resulting in cell since nutrition supply is limited makes its own division
Breeding is limited.
Therefore, a kind of debubbling method convenient, effective during photo-biological cell adheres to and cultivates is developed to photo-biological
The attachment culture of cell is of great significance.
Summary of the invention
In consideration of it, to solve problem of the prior art, the embodiment of the present invention provides a kind of culture side of photo-biological cell
Method can convenient, effectively eliminate the bubble in the attachment culture of photo-biological cell, further increase its reproductive efficiency.
In order to achieve the above objectives, the embodiment of the present invention adopts the following technical scheme that
The embodiment of the invention provides a kind of cultural methods of photo-biological cell, and the method includes steps (1), judgement
Bubble whether there is between carrier layer and cloth liquid layer, or be present between the carrier layer and cellular layer；Step (2), basis
The position that the bubble occurs, adjusts the default supply amount of culture solution, until the bubble is fully drained；Including when described
When bubble is present between the carrier layer and the cloth liquid layer, increase the supply amount of the culture solution, until the bubble quilt
It is completely exhausted out；When the bubble is present between the carrier layer and the cellular layer, stop, or stop the culture repeatedly
The supply of liquid, until the bubble is fully drained；Step (3), the default supply amount according to the culture solution, continue to train
Support the photo-biological cell.
Optionally, the step (1) specifically includes, when protrusion occurs in cell layer surface, if the protrusion is in individual point
Cloth, and the photo-biological cell in the protrusion is in the photo-biological cellular colours in drying regime, and/or the protrusion
It changes, it is determined that the bubble is present between carrier layer and cloth liquid layer；When protrusion occurs in cell layer surface, if described
Protrusion is in population distribution, and the photo-biological cell in the protrusion is in the light on moisture state, and/or the protrusion
Biological cell color does not change, it is determined that the bubble is present between carrier layer and cellular layer.
Optionally, described when the bubble is present between the carrier layer and the cloth liquid layer, increase the culture
The supply amount of liquid specifically includes, until the bubble is fully drained when the bubble is present in the carrier layer and the cloth
When between liquid layer, increase the supply amount of the culture solution, the culture solution made to pass through 1~20 process on the cloth liquid layer,
So that the bubble is fully drained；Wherein, the flow definition is to be flowed to the culture solution from the top of the cloth liquid layer
The process of bottom end.
Optionally, described when the bubble is present between the carrier layer and the cellular layer, stop, or stop repeatedly
The only supply of the culture solution specifically includes until the bubble is fully drained, and step (2.1) is present in when the bubble
When between the carrier layer and the cellular layer, stop the supply of the culture solution；Step (2.2), the observation carrier layer with
The elimination situation of the bubble between the cellular layer illustrates the bubble quilt if not observing the presence of the bubble
It is completely exhausted out；Step (2.3) if, between the carrier layer and the cellular layer there are still the bubble not being eliminated,
Restart to supply the culture solution until there is the continuous drop of the culture solution to flow out from the cloth liquid layer, stopping supplies again
To the culture solution；Step (2.4), circulation successively execute the step (2.2) and the step (2.3), until the bubble
It is fully drained.
Optionally, described when the bubble is present between the carrier layer and the cellular layer, stop, or stop repeatedly
The only supply of the culture solution, until the bubble is fully drained, it further include that the bubble of discharge is moved to the carrier
Between layer and the cloth liquid layer；Increase the supply amount of the culture solution, until the bubble is fully drained.
Optionally, increase the supply amount of the culture solution to not sweeping away the photo-biological cell in the carrier layer
Maximum stream flow.
Optionally, the default supply amount according to the culture solution continues to cultivate the photo-biological cell, packet
It includes, defoaming agent is added in Xiang Suoshu culture solution, or remove the light being dissolved in the culture solution through film using selectivity
The cell secreta of biological cell.
Optionally, before executing the step (3), the method also includes checking the carrier layer and the cloth liquid
Whether the bubble between layer, between the carrier layer and the cellular layer thoroughly excludes；If the carrier layer and the cloth
There are still the bubble then repeating said steps (2) between liquid layer and/or between the carrier layer and the cellular layer.
It is further preferred that between the inspection carrier layer and the cloth liquid layer, the carrier layer and the cell
Whether the bubble between layer thoroughly excludes, if between the carrier layer and the cloth liquid layer and/or the carrier layer and institute
It states between cellular layer there are still the bubble then repeating said steps (2), specifically includes, gradually decrease the supply of the culture solution
Until stopping completely, observe whether the carrier layer is bonded completely with the cloth liquid layer, the carrier layer with the cellular layer is
No complete fitting；If the carrier layer be not exclusively bonded with the cloth liquid layer, and/or the carrier layer and the cellular layer it is endless
Full fitting, then illustrate between the carrier layer and the cloth liquid layer and/or between the carrier layer and the cellular layer there are still
The bubble, repeating said steps (2).
Optionally, described according to the default supply amount, continue to cultivate the photo-biological cell, including, when the bubble
When being present between the carrier layer and the cloth liquid layer, the supply amount of the culture solution is gradually decreased to the default supply
Amount, continues to cultivate the photo-biological cell；When the bubble is present between the carrier layer and the cellular layer, gradually increase
Add the supply amount of the culture solution to the default supply amount, continues to cultivate the photo-biological cell.
Optionally, before executing the step (1), the method also includes training to the photo-biological cell
At the beginning of supporting, the bubble generated due to adhering to the operation of culture between carrier layer and cloth liquid layer is removed.
It is further preferred that described at the beginning of cultivating the photo-biological cell, removal carrier layer and cloth liquid layer it
Between due to attachment culture operation and the bubble that generates, specifically include, by a line of carrier layer side corresponding with cloth liquid layer
It is in contact；To be in contact while as starting point, make the carrier layer and the cloth liquid layer to opposite with the starting point side another
Side slowly sequentially contacts；And/or after carrier layer is placed on cloth liquid layer surface, the supply amount of culture solution and/or water is adjusted
Greater than the default supply amount of the culture solution, to remove the behaviour cultivated due to attachment between the carrier layer and the cloth liquid layer
The bubble made and generated；Stop the supply of the culture solution and/or the water, so that the carrier layer and the cloth liquid layer are close
Fitting；The supply amount of the culture solution is gradually increased until the default supply amount.
It is further preferred that the supply amount of the adjusting culture solution and/or water is greater than the default supply amount of the culture solution
To the maximum stream flow not swept away the photo-biological cell in the carrier layer.
Based on this, a kind of cultural method of photo-biological cell is provided through the embodiment of the present invention, when carrier layer and cloth liquid
After there is bubble between layer, by increasing the supply amount of culture solution, so that bubble follows the flowing of culture solution to be expelled to load together
Except body layer and cloth liquid layer, thus the bubble between two layers of removal；After occurring bubble between carrier layer and cellular layer, by stopping
Only, or repeatedly the supply for stopping culture solution, so that the bubble being present between carrier layer and cellular layer is by atmospheric pressure and culture
The collective effect for the vacuum force that liquid is formed between cloth liquid layer and carrier layer, by being moved between carrier layer and cellular layer or
Drain between carrier layer and cloth liquid layer, can be flowed out later with the culture solution for increasing flow, thus remove carrier layer and cellular layer it
Between bubble.Achieve the effect that convenient, effectively elimination photo-biological cell adheres to the bubble in culture, it is numerous to further improve its
Grow efficiency.
Detailed description of the invention
In order to more clearly explain the embodiment of the invention or the technical proposal in the existing technology, to embodiment or will show below
There is attached drawing needed in technical description to be briefly described, it should be apparent that, the accompanying drawings in the following description is only this
Some embodiments of invention for those of ordinary skill in the art without creative efforts, can be with
It obtains other drawings based on these drawings.
Fig. 1 is the schematic illustration of current photo-biological cell attachment culture；
Fig. 2 is a kind of cultural method flow diagram of photo-biological cell provided in an embodiment of the present invention；
Fig. 3 is the operation chart that carrier layer is in contact with cloth liquid layer in the embodiment of the present invention.
Specific embodiment
Following will be combined with the drawings in the embodiments of the present invention, and technical solution in the embodiment of the present invention carries out clear, complete
Site preparation description, it is clear that described embodiments are only a part of the embodiments of the present invention, instead of all the embodiments.It is based on
Embodiment in the present invention, it is obtained by those of ordinary skill in the art without making creative efforts every other
Embodiment shall fall within the protection scope of the present invention.
As shown in Fig. 2, the embodiment of the invention provides a kind of cultural method of photo-biological cell, this method includes,
Step (1) judges that bubble whether there is between carrier layer and cloth liquid layer, or be present in carrier layer and cellular layer it
Between；
Step (2), the position occurred according to bubble, adjust the default supply amount of culture solution, until bubble is fully drained；
Including when bubble is present between carrier layer and cloth liquid layer, increasing the supply amount of culture solution, until bubble is fully drained；
When bubble is present between carrier layer and cellular layer, stop, or stop the supply of culture solution repeatedly, until bubble is arranged completely
Out；
Step (3), the default supply amount according to culture solution continue to cultivate photo-biological cell.
It should be noted that above-mentioned photo-biological cell can be spherical algae, filamentous algae, arc algae, spirulina, triangle
Any one of algae, polygonal algae, scenedesmus, chlorella, Chlorococcum, haematococcus, silkweed and filamentous cyanobacteria.
In embodiments of the present invention, cloth liquid layer is by having material that hold liquid ability, such as porous or water wetted material to constitute, with true
Protecting culture solution can adequately be absorbed during the direction along cell bottom to surface layer is permeated by cell.
When protrusion occurs in photo-biological cell surface, then there is bubble in the system of above-mentioned attachment culture in explanation, specifically
Observation process can be completed by human eye, cultivating system can also be monitored by setting up high-definition camera.
In above-mentioned steps (2), the default supply amount of culture solution refers to original supply amount when normal breeding culture.It is above-mentioned
The supply of culture solution " repeatedly stop " refer to and stop culture solution supply, open culture solution supply, stop the anti-of culture solution supply
Multiple process.
Based on this, a kind of cultural method of photo-biological cell is provided through the embodiment of the present invention, when carrier layer and cloth liquid
After there is bubble between layer, by increasing the supply amount of culture solution, so that bubble follows the flowing of culture solution to be expelled to load together
Except body layer and cloth liquid layer, thus the bubble between two layers of removal；After occurring bubble between carrier layer and cellular layer, by stopping
Only, or repeatedly the supply for stopping culture solution, so that the bubble being present between carrier layer and cellular layer is by atmospheric pressure and culture
The collective effect for the vacuum force that liquid is formed in cloth liquid layer and carrier interlayer, by being moved to or arranging between carrier layer and cellular layer
To between carrier layer and cloth liquid layer, can be flowed out later with the culture solution for increasing flow, to remove between carrier layer and cellular layer
Bubble.Achieve the effect that convenient, effectively elimination photo-biological cell adheres to the bubble in culture, further improves its breeding
Efficiency.
Each step and its concrete operations during an embodiment of the present invention will be described in detail below:
Preliminary step
Before executing step (1), above-mentioned cultural method provided in an embodiment of the present invention may also include, thin to photo-biological
At the beginning of born of the same parents cultivate, the bubble generated due to adhering to the operation of culture between carrier layer and cloth liquid layer is removed.
Here, above-mentioned " at the beginning of culture " refers to, after in photo-biological cell inoculation to carrier layer, culture solution supply is opened
Before beginning normally to cultivate.
As shown in figure 3, above-mentioned steps specifically can be used the combination of any one of following two mode or two ways into
Row, it may be assumed that
(1) mode one
The a line of carrier layer side corresponding with cloth liquid layer is in contact；It (is marked in figure while as starting point using being in contact
To contact carrier layer slowly sequentially to the another a line (in figure label be) opposite with starting point side with cloth liquid layer, with to the greatest extent
Amount avoids having bubble folder between the two layers during this.
(2) after carrier layer is placed on cloth liquid layer surface by mode two, the supply amount for adjusting culture solution and/or water is greater than culture
The default supply amount of liquid, the bubble generated with removing the operation cultivated due to attachment between carrier layer and cloth liquid layer；Stop
The supply of culture solution and/or water, so that carrier layer is fitted closely with cloth liquid layer.
Here, the supply amount for adjusting culture solution and/or water is greater than the default supply amount of culture solution to will not be in carrier layer
The maximum stream flow that photo-biological cell is swept away, in order to avoid the photo-biological cell in inoculation is washed away.
I.e. according to after the operation of aforesaid way one between the two layers still with the presence of bubble, then mode two can be continued to execute
Operation.
The supply amount of culture solution can be gradually increased after having carried out aforesaid way one and/or mode two until default supply
Amount.
Step (1)
Further, above-mentioned steps (1) specifically may include following process:
When protrusion occurs in cell layer surface, if protrusion is in individual distribution, and the photo-biological cell in protrusion is in dry shape
Photo-biological cellular colours in state, and/or protrusion change, such as can be cellular colours flavescence, it is determined that bubble exists
Between carrier layer and cloth liquid layer；
When protrusion occurs in cell layer surface, if protrusion is in population distribution, and the photo-biological cell in protrusion is in wet shape
Photo-biological cellular colours in state, and/or protrusion do not change, it is determined that bubble is present between carrier layer and cellular layer.
Wherein, above-mentioned " individual distribution " refers to that negligible amounts, the individual of bubble are larger, and multiple bubbling polymerizations form, are in
The sheet bubble of protuberance；Above-mentioned " population distribution " refers to that the quantity of bubble is more, individual smaller, how hemispherical or most
Spherical a group.
Step (2)
When bubble is present between carrier layer and cloth liquid layer, increase the supply amount of culture solution, until bubble is arranged completely
Out the step of, specifically includes:
When bubble is present between carrier layer and cloth liquid layer, increase the supply amount of culture solution, makes culture solution in cloth liquid layer
It is upper to pass through 1~20 process, so that bubble is fully drained；Wherein, flow definition is to be flowed with culture solution from the top of cloth liquid layer
To the process of bottom end.
Here, comprehensively consider production efficiency, culture solution passes through 5~10 processes preferably on cloth liquid layer.Also, increase
The supply amount of culture solution is to the maximum stream flow not swept away the photo-biological cell in carrier layer, in order to avoid influence subsequent normal breeding
Culture.
When bubble is present between carrier layer and cellular layer, stop, or stop the supply of culture solution repeatedly, until bubble
The step of being fully drained specifically includes:
Step (2.1), when bubble is present between carrier layer and cellular layer, stop the supply of culture solution；Here, it cultivates
After liquid stops supply, the culture solution that iuntercellular fills gradually is lost, and can be clearly observed cellular layer at this time by moistening shape
State becomes drying regime, can't see the reflective of the liquid of iuntercellular culture solution.
The elimination situation of bubble between step (2.2), observation carrier layer and cellular layer, if not observing depositing for bubble
Then illustrating that bubble is fully drained；It can continue above-mentioned step (3) after this.
Step (2.3) if, there are still the bubbles not being eliminated between carrier layer and cellular layer, restart supply training
Nutrient solution is until there is the continuous drop of culture solution to flow out from cloth liquid layer, and stopping supplies culture solution again；At this point, cell is by aforementioned step
Suddenly the drying regime in (2.1) becomes moisture state, and finally becomes drying regime again.
Step (2.4), circulation successively execute step (2.2) and step (2.3), up to bubble is fully drained, herein it
It can continue above-mentioned step (3) afterwards.
During carrying out above-mentioned steps (2.1)~step (2.4), the bubble of discharge can be moved to carrier layer and cloth liquid
Between layer.The step of bubble between above-mentioned removal carrier layer and cloth liquid layer can be performed later.
Wherein, during removing the bubble between carrier layer and cloth liquid layer, increase the supply amount of culture solution to not by load
The maximum stream flow that photo-biological cell on body layer is swept away, in order to avoid influence subsequent normal breeding culture.
Checking step
Further, before executing step (3), the above method further include:
Check whether the bubble between carrier layer and cloth liquid layer, between carrier layer and cellular layer thoroughly excludes；If bubble is thorough
Bottom excludes then to can proceed with subsequent step (3).
If between carrier layer and cloth liquid layer, and/or between carrier layer and cellular layer, there are still bubbles to repeat step (2).
Above-mentioned checking process specifically comprises the following steps:
Gradually decrease culture solution supply until completely stop, observation carrier layer whether be bonded completely with cloth liquid layer, carrier
Whether layer is bonded completely with cellular layer；
If carrier layer is not exclusively bonded with cloth liquid layer, and/or carrier layer is not exclusively bonded with cellular layer, illustrate carrier layer
There are still bubbles between cloth liquid layer and/or between carrier layer and cellular layer, repeat step (2).
Here it is possible to which the whether complete of fitting judged with the presence or absence of protrusion by observation cell surface.
Step (3)
After having executed above-mentioned steps (2), if bubble is when being present between carrier layer and cloth liquid layer, can to gradually decrease culture
The supply amount of liquid continues to cultivate photo-biological cell to default supply amount；If bubble is when being present between carrier layer and cellular layer,
The supply amount of culture solution can be gradually increased to default supply amount, continue to cultivate photo-biological cell.
Further, since the cell secreta of photo-biological cell is easily blistered, above-mentioned steps (3) " continue to train
Support " it further include that defoaming agent is added into culture solution, or it is thin through the photo-biological that is dissolved in culture solution of film removal using selectivity
The cell secreta of born of the same parents, to alleviate the bubble problem in subsequent attachment culture.
Wherein, selectivity is specifically as follows the UF membranes facility such as ultrafiltration membrane, microfiltration membranes through film.
The above method that two specific embodiments are provided with the present invention is described in detail embodiment is provided below.
Embodiment 1
Using composite fibre film as carrier layer in the present embodiment, using canvas as cloth liquid layer, using lucite tube to cloth liquid layer
Supply culture solution；In the collecting tank (volume is 1m × 2m × 0.2m) that culture solution is stored in, the high 0.08m of liquid level.By small-sized latent
Water pump supplies the culture solution in collecting tank to lucite tube, on the pipeline of connection pump and lucite tube, is provided with stream
Adjustable valve.
Specific incubation step is as described below:
Bottle green scenedesmus is seeded on composite fibre film, protrusion occurs in cell layer surface after culture 2 days, it is observed that hair
Existing: protrusion is about 2cm in individual distribution (quantity totally 3), in irregular shape, raised floor space minimum2, be about 10cm2
(illustrating that raised individual is larger), convex surfaces cell is in yellowish-brown, judges bubble between carrier layer and cloth liquid layer；
Increase about 15 processes of culture solution flow control valve (having no that cell falls off from carrier layer during this), by flow
Regulating valve is recalled to default supply amount (i.e. the initial cultivation flow of culture solution).Cell layer surface is observed without protrusion, illustrates bubble
Excluded completely；
2 drop polyethers foam inhibitors are instilled into culture solution, continue " attachment culture ".
Embodiment 2
Using cellulose acetate film as carrier layer in the present embodiment, using sponge as cloth liquid layer, using lucite tube to cloth liquid layer
Supply culture solution；In the collecting tank (volume is 1m × 2m × 0.2m) that culture solution is stored in, the high 0.10m of liquid level.By small-sized latent
Water pump supplies the culture solution in collecting tank to lucite tube, on the pipeline of connection pump and lucite tube, is provided with stream
Adjustable valve.
Specific incubation step is as described below:
Blue-green spirulina is seeded on cellulose acetate film, protrusion occurs in cell layer surface after culture 1 day, it is observed that hair
Existing: it is about 0.2cm that protrusion, which is mostly dome-type, raised floor space minimum in population distribution (quantity totally 135), shape,2, the largest of about
For 2cm2(illustrating that raised individual is smaller), convex surfaces cell is in blue-green, judges bubble between cellular layer and carrier layer；
It closes culture solution flow control valve to spirulina surface cell culture to be lost, i.e., cell is in dry shape, and observation is thin
Cellular surface protrusion eliminates about 70%~80%, and turn-on flow rate regulating valve (has no that cell falls off from carrier layer during this).When
The continuous drop of culture solution has been observed after flowing out in cloth liquid layer, has been again switched off flow control valve to spirulina superficial cell in dry
After dry shape, it has been observed that cell surface protrusion is all eliminated, illustrate that bubble is excluded completely；
The culture solution in collecting tank is filtered one time using ultrafiltration apparatus, to remove cell secreta therein, dissolved matter
And after impurity, continue " attachment culture ".
It should be noted that those of ordinary skill in the art will appreciate that: realize whole or the portion of above method embodiment
This can be accomplished by hardware associated with program instructions step by step, and program above-mentioned can store in a computer-readable storage
In medium, which when being executed, executes step including the steps of the foregoing method embodiments；And storage medium above-mentioned may include:
ROM (Read-Only Memory, the abbreviation of read-only memory), RAM (Random Access Memory, random access memory
The abbreviation of device) the various media that can store program code such as magnetic or disk.
The above description is merely a specific embodiment, but scope of protection of the present invention is not limited thereto, any
Those familiar with the art in the technical scope disclosed by the present invention, can easily think of the change or the replacement, and should all contain
Lid is within protection scope of the present invention.Therefore, protection scope of the present invention should be based on the protection scope of the described claims.
Claims (11)
1. a kind of cultural method of photo-biological cell, which is characterized in that the method includes,
Step (1) judges that bubble whether there is between carrier layer and cloth liquid layer, or be present in the carrier layer and cellular layer it
Between；
Step (2), the position occurred according to the bubble, adjust the default supply amount of culture solution, until the bubble is complete
Discharge；Including increasing the supply of the culture solution when the bubble is present between the carrier layer and the cloth liquid layer
Amount, until the bubble is fully drained；When the bubble is present between the carrier layer and the cellular layer, stop,
Or stop the supply of the culture solution repeatedly, until the bubble is fully drained；
Step (3), the default supply amount according to the culture solution, continue to cultivate the photo-biological cell；
Wherein, the step (1) includes,
When cell layer surface appearance protrusion, if the protrusion is distributed in individual, and the photo-biological cell in the protrusion
It changes in the photo-biological cellular colours in drying regime, and/or the protrusion, it is determined that the bubble is present in load
Between body layer and cloth liquid layer；
When cell layer surface occur protrusion when, if it is described protrusion be in population distribution, and it is described protrusion on the photo-biological cell
It does not change in the photo-biological cellular colours on moisture state, and/or the protrusion, it is determined that the bubble is present in
Between carrier layer and cellular layer；
In the step (2),
The supply for stopping culture solution repeatedly refers to, stops culture solution supply, opens culture solution supply, stops culture solution supply
Repetitive process；
The step (3) includes,
When the bubble is present between the carrier layer and the cloth liquid layer, the supply amount of the culture solution is gradually decreased extremely
The default supply amount continues to cultivate the photo-biological cell；
When the bubble is present between the carrier layer and the cellular layer, the supply amount of the culture solution is gradually increased extremely
The default supply amount continues to cultivate the photo-biological cell.
2. cultural method according to claim 1, which is characterized in that it is described when the bubble be present in the carrier layer with
When between the cloth liquid layer, increase the supply amount of the culture solution, until the bubble is fully drained, specifically include,
When the bubble is present between the carrier layer and the cloth liquid layer, increases the supply amount of the culture solution, make institute
Culture solution is stated on the cloth liquid layer by 1~20 process, so that the bubble is fully drained；Wherein, the process is fixed
Justice is to flow to the process of bottom end from the top of the cloth liquid layer with the culture solution.
3. cultural method according to claim 1, which is characterized in that it is described when the bubble be present in the carrier layer with
When between the cellular layer, stop, or stop the supply of the culture solution repeatedly, until the bubble is fully drained, specifically
Including,
Step (2.1), when the bubble is present between the carrier layer and the cellular layer, stop the confession of the culture solution
It gives；
The elimination situation of the bubble between step (2.2), the observation carrier layer and the cellular layer, if not observing institute
The presence of bubble is stated, then illustrates that the bubble is fully drained；
Step (2.3) if, between the carrier layer and the cellular layer there are still the bubble not being eliminated, open again
Begin to supply the culture solution until there is the continuous drop of the culture solution to flow out from the cloth liquid layer, again described in stopping supply
Culture solution；
Step (2.4), circulation successively execute the step (2.2) and the step (2.3), until the bubble is arranged completely
Out.
4. cultural method according to claim 1, which is characterized in that it is described when the bubble be present in the carrier layer with
When between the cellular layer, stop, or stop the supply of the culture solution repeatedly, until the bubble is fully drained, also wraps
It includes,
The bubble of discharge is moved between the carrier layer and the cloth liquid layer；
Increase the supply amount of the culture solution, until the bubble is fully drained.
5. cultural method according to claim 1 or 4, which is characterized in that increase the supply amount of the culture solution to
The maximum stream flow that the photo-biological cell in the carrier layer is swept away.
6. cultural method according to claim 1, which is characterized in that the default supply according to the culture solution
Amount, continues to cultivate the photo-biological cell, including,
Defoaming agent is added into the culture solution, or removes the light being dissolved in the culture solution through film using selectivity
The cell secreta of biological cell.
7. cultural method according to claim 1, which is characterized in that before executing the step (3), the method is also
Including,
Check whether the bubble between the carrier layer and the cloth liquid layer, between the carrier layer and the cellular layer is thorough
Bottom excludes；
If between the carrier layer and the cloth liquid layer and/or between the carrier layer and the cellular layer, there are still the gas
Steep then repeating said steps (2).
8. cultural method according to claim 7, which is characterized in that it is described check the carrier layer and the cloth liquid layer it
Between, the bubble between the carrier layer and the cellular layer whether thoroughly exclude, if the carrier layer and the cloth liquid layer
Between and/or the carrier layer and the cellular layer between there are still the bubble then repeating said steps (2), specifically include,
The supply of the culture solution is gradually decreased until stopping completely, observes whether the carrier layer pastes completely with the cloth liquid layer
It closes, whether the carrier layer is bonded completely with the cellular layer；
If the carrier layer is not exclusively bonded with the cloth liquid layer, and/or the carrier layer is not exclusively bonded with the cellular layer,
Then illustrate between the carrier layer and the cloth liquid layer, and/or there are still the gas between the carrier layer and the cellular layer
Bubble, repeating said steps (2).
9. cultural method according to claim 1, which is characterized in that before executing the step (1), the method is also
Including,
At the beginning of cultivating the photo-biological cell, the operation cultivated due to attachment between carrier layer and cloth liquid layer is removed
And the bubble generated.
10. cultural method according to claim 9, which is characterized in that described to be cultivated to the photo-biological cell
At the beginning of, the bubble generated due to adhering to the operation of culture between carrier layer and cloth liquid layer is removed, is specifically included,
The a line of carrier layer side corresponding with cloth liquid layer is in contact；To be in contact while as starting point, make the carrier layer
It is slowly sequentially contacted with the cloth liquid layer to the another a line opposite with the starting point side；And/or carrier layer is placed on cloth
Behind liquid layer surface, the supply amount of adjusting culture solution and/or water is greater than the default supply amount of the culture solution, to remove the carrier
The bubble generated due to adhering to the operation of culture between layer and the cloth liquid layer；Stop the culture solution and/or the water
Supply so that the carrier layer is fitted closely with the cloth liquid layer；
The supply amount of the culture solution is gradually increased until the default supply amount.
11. cultural method according to claim 10, which is characterized in that the supply amount for adjusting culture solution and/or water
Greater than the culture solution default supply amount to the maximum stream flow not swept away the photo-biological cell in the carrier layer.
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| CN204509276U (en) * | 2015-03-25 | 2015-07-29 | 新奥科技发展有限公司 | A kind of bioreactor |
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| US6348347B1 (en) * | 1998-03-31 | 2002-02-19 | Micro Gaia Co., Ltd. | Fine algae culture device |
| US7897379B2 (en) * | 2007-02-26 | 2011-03-01 | Corning Incorporated | Device and method for reducing bubble formation in cell culture |
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