CN105973680A - Preparation method for algae spot samples, algae spot samples and method for enhancing microalgae P700 signals - Google Patents
Preparation method for algae spot samples, algae spot samples and method for enhancing microalgae P700 signals Download PDFInfo
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- CN105973680A CN105973680A CN201610594026.2A CN201610594026A CN105973680A CN 105973680 A CN105973680 A CN 105973680A CN 201610594026 A CN201610594026 A CN 201610594026A CN 105973680 A CN105973680 A CN 105973680A
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- algae
- sample
- speckle
- preparation
- microalgae
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
Abstract
The invention discloses a preparation method for algae spot samples, the algae spot samples and a method for enhancing microalgae P700 signals. The preparation method for the algae spot samples comprises the steps that firstly, microalgae suspension liquid is dropped to a filter membrane until a preset concentration is reached; secondly, algae spots are formed after filtration is carried out through the filter membrane, and the algae spot samples with the diameters not smaller than the diameter of a probe are acquired; thirdly, the algae spot samples are arranged in a colorless and transparent bag; fourthly, after air in the colorless and transparent bag containing the algae spot samples is discharged, sealing treatment is carried out. By means of the preparation method, the microalgae suspension liquid can be prepared into the algae spot samples, the intensity of the P700 signals can be greatly improved, the signal noise can be lowered, and the advantages that operation is easy and convenient, the cost is low and time is saved are achieved.
Description
Technical field
The present invention relates to instrument test technical field biology, particularly relate to the preparation side of a kind of algae speckle sample
Method and algae speckle sample and the method strengthening microalgae P700 signal.
Background technology
Photosynthesis is most important chemical reaction on the earth, and its process is extremely complex.Photosynthetic
Measure and research always plant physiology, phytoecology, agronomy, forestry, gardening, algae
The study hotspot in the fields such as physiological ecology.
Photosynthesis substantially can be divided into photoreaction and dark reaction two parts.Wherein photoreaction occurs at class capsule
On body film, it is made up of multiple photoreaction associated components, mainly has Photosystem I and Photosystem I I.P700
It is the photosynthetic reaction center of Photosystem I, is made up of chlorophyll and conjugated protein, because it is at 700nm
There is absworption peak at place so gaining the name.Owing to P700 plays the electronics of key during photosynthesis is carried out
Transmission effect, it is therefore desirable to its state is measured.P700 is in reduction-state when obtaining electronics, loses
The light of 700nm is absorbed by the P700 being in during electronics under oxidation state, two states subtle difference.
Therefore can be by measuring the slight change that 700nm light is absorbed by sample, namely P700 signal, comes
Understand the redox state of P700, thus understand its effect played in photosynthesis.
The difference absorbed 700nm light due to the oxidized and reduced of P700 is the most small, therefore
P700 measuring instrument needs the highest sensitivity, when the chlorophyll content of sample is higher simultaneously
Obvious P700 signal just can be detected, if chlorophyll content is low, then be difficult to detection.Therefore,
The material higher to chlorophyll content, such as plant leaf blade etc., is easier to obtain good P700 signal.
But high for transparency, for the thin microalgae suspension that absorbance is the lowest, P700 signal will be non-
The faintest, add frustule and constantly move in suspension, cause signal big ups and downs, it is impossible to be full
The measurement of foot P700 needs.
Prior art there is problems in that
The thinnest micro algae growth early stage and dense micro algae growth latter stage, its photosynthetic activity and
There is greatest differences in feature.Only measure the P700 signal in micro algae growth latter stage cannot reflect frequently more
The photosynthesis characteristics of important micro algae growth early stage.This makes microalgae P700 measurement data there is the serious flaw
Defect.
2. microalgae is even several weeks from the training of thin growth early stage to dense growth need a couple of days in latter stage
The time of supporting, elapsed time is long.
3. even if, in the micro algae growth latter stage that algae cell density is higher, its chlorophyll concentration is still far below leaf
The high chlorophyll samples such as sheet, therefore P700 signal is the most weak.Additionally frustule is in suspension
Motion, also can cause the biggest interference, it is impossible to obtain preferable P700 signal to P700 signal.
The P700 of microalgae sample measures technology, it has also become limit the bottleneck of microalgae photosynthesis research,
Also had a strong impact on the P700 measurement of correlation instrument using value in algae photosynthesis field, be badly in need of
A kind of simple and effective method strengthening microalgae P700 signal.
Summary of the invention
It is an object of the present invention to provide preparation method and the algae speckle sample of a kind of algae speckle sample and strengthen microalgae
The method of P700 signal, makes microalgae suspension algae speckle sample, can increase substantially the letter of P700
Number intensity, reduces signal noise, has advantage easy and simple to handle, with low cost, time saving.
The technical scheme that the present invention provides is as follows:
A kind of preparation method of algae speckle sample, including:
Step S10 instills microalgae suspension on filter membrane, reaches preset concentration；
Step S10 described in step S20 forms algae speckle after membrane filtration, obtains diameter not less than probe diameter
Algae speckle sample.
It is further preferred that also include:
Described algae speckle sample is arranged in water white transparency bag by step S30.
It is further preferred that also include:
After described step S30 is provided with the water white transparency bag air-out of algae speckle sample by step S40, carry out
Encapsulation process.
It is further preferred that include:
Described filter membrane also includes: glass fiber filter, or qualitative filter membrane.
It is further preferred that include:
Described filter membrane diameter is set to 40-50mm.
It is further preferred that include:
Described filter membrane diameter is set to 47mm.
It is further preferred that also include:
Described step S10 forms algae speckle after membrane filtration, obtains the diameter algae speckle sample more than 1.5cm.
It is further preferred that also include:
Step S10 instills microalgae suspension on filter membrane, for reaching preset concentration, implements the most repeatedly to instill
Described microalgae suspension.
The present invention also provides for a kind of algae speckle sample, the preparation method of a kind of algae speckle sample the algae speckle sample prepared
Product.
The present invention also provides for a kind of method testing P700 signal, including:
The described test sample of preparation is placed on the test table in P700 measuring instrument by step S10；
Test sample described in step S20 is placed between receiving transducer and the transmitting probe of described test table；
Step S30 records the data message of described receiving transducer.
The preparation method of a kind of algae speckle sample provided by the present invention and algae speckle sample and strengthen microalgae P700
The method of signal, it is possible to bring at least one beneficial effect following:
1, the invention provides the preparation method of algae speckle sample, substantially increase frustule density and
Chlorophyll concentration, thus it is effectively increased the measurement signal of P700, solve microalgae in prior art
The problem of P700 weak output signal.
2, the invention provides the preparation method of algae speckle sample, it is to avoid microalgae cell is in suspension
The big ups and downs of P700 signal that caused of motion, thus be effectively increased stablizing of P700 signal
Property, solve the problem that in prior art, microalgae P700 measures signal big ups and downs.
3, the present invention is for the thinnest microalgae sample, such as thin micro algae growth early stage suspension
Or the thin microalgae sample etc. in natural water, by the way of the most repeatedly instilling filter membrane, still can be by
Microalgae cell is enriched to preset concentration, which greatly enhances the dense of thin microalgae suspension P700 measurement
Degree lower limit.For the thinnest microalgae suspension sample, the present invention is equally applicable.
4, algae speckle sample is arranged in water white transparency bag by the present invention, carries out at sealing after air-out
Reason, it is to avoid the photosynthetic activity of microalgae sample is interfered because of moisture loss, it is ensured that microalgae sample
P700 signal accurate.
5, applied widely to microalgae concentration of the present invention.Before being not only applicable to thin micro algae growth
Phase sample, is also applied for dense micro algae growth late-run sample.By the method for the invention, permissible
The P700 of the microalgae sample realizing different growing stage measures, and improves the comparability of measurement result.
6, the invention provides the preparation method of algae speckle sample, can be rich by microalgae cell at short notice
Collection is to preset concentration.Compared with prior art, it is not necessary to the cultivation through even several weeks a couple of days is bred, greatly
Save greatly the time.
7, the present invention is simple for sample production method, and the cycle is short, simultaneously for preserving algae speckle sample
Activity, uses sealing bag to be packaged, with low cost；Environment is not resulted in pollution.
8, the present invention solves microalgae sample and cannot obtain the technical barrier of preferable P700 signal, breaks through
Limit the bottleneck of microalgae photosynthesis research, P700 measurement of correlation instrument is greatly improved in algae
The using value in photosynthesis field.
Accompanying drawing explanation
Below by the way of the most understandable, accompanying drawings preferred implementation, to a kind of algae speckle sample
Preparation method and algae speckle sample and strengthen the above-mentioned characteristic of method of microalgae P700 signal, technical characteristic, excellent
Point and implementation thereof are further described.
Fig. 1 is the flow chart of the embodiment of the preparation method of the present invention a kind of algae speckle sample；
Fig. 2 is another flow chart of the embodiment of the preparation method of the present invention a kind of algae speckle sample；
Fig. 3 is the flow chart of the embodiment of a kind of method testing P700 signal of the present invention；
Fig. 4 is another embodiment signal graph of a kind of method testing P700 signal of the present invention；
Fig. 5 is another embodiment signal graph of a kind of method testing P700 signal of the present invention；
Fig. 6 is another embodiment signal graph of a kind of method testing P700 signal of the present invention.
Detailed description of the invention
In order to be illustrated more clearly that the embodiment of the present invention or technical scheme of the prior art, will compare attached below
The detailed description of the invention of the figure explanation present invention.It should be evident that the accompanying drawing in describing below is only the present invention
Some embodiments, for those of ordinary skill in the art, on the premise of not paying creative work,
Other accompanying drawing can also be obtained according to these accompanying drawings, and obtain other embodiment.
For making simplified form, only schematically show part related to the present invention in each figure, they are also
Do not represent its practical structures as product.It addition, so that simplified form readily appreciates, have in some figure
There are the parts of identical structure or function, only symbolically depict one of them, or only marked therein
One.In this article, " one " not only represents " only this ", it is also possible to represent the situation of " more than one ".
Photosynthesis substantially can be divided into photoreaction and dark reaction two parts.Wherein photoreaction occurs at thylakoid membrane
On, it is made up of multiple photoreaction associated components, mainly has Photosystem I and Photosystem I I.P700 is Photosystem I
Photosynthetic reaction center, be made up of chlorophyll and conjugated protein, because of its at 700nm, have absworption peak so
Gain the name.Owing to P700 plays the Electron Transfer of key during photosynthesis is carried out, it is therefore desirable to right
Its state is measured.P700 is in reduction-state when obtaining electronics, be in oxidation state, two states during betatopic
Under P700 the light of 700nm absorbed have subtle difference.Therefore by measuring sample, 700nm light can be inhaled
The slight change received, namely P700 signal, understand the redox state of P700, thus understand its
Effect played in photosynthesis.
The difference absorbed 700nm light due to the oxidized and reduced of P700 is the most small, and therefore P700 surveys
Measuring appratus needs the highest sensitivity, just can detect bright when the chlorophyll content of sample is higher simultaneously
Aobvious P700 signal, if chlorophyll content is low, is then difficult to detection.Therefore, higher to chlorophyll content
Material, such as plant leaf blade etc., is easier to obtain good P700 signal.But high for transparency, absorbance
For the lowest thin microalgae suspension, P700 signal will be the faintest, adds frustule at suspension
In constantly move, cause signal big ups and downs, it is impossible to meet the measurement needs of P700.
The present invention provides an embodiment of the preparation method of a kind of algae speckle sample, with reference to shown in Fig. 1；Including:
Step S10 instills microalgae suspension on filter membrane, reaches preset concentration；
Step S10 described in step S20 forms algae speckle after membrane filtration, obtains diameter not less than probe diameter
Algae speckle sample.
Concrete, the difference absorbed 700nm light due to the oxidized and reduced of P700 is the most micro-
Little, therefore P700 measuring instrument needs the highest sensitivity, and the chlorophyll of sample contains simultaneously
Obvious P700 signal just can be detected when measuring higher, if chlorophyll content is low, then be difficult to detection.
Therefore, the material higher to chlorophyll content, such as plant leaf blade etc., it is easier to obtain good P700
Signal.But high for transparency, for the thin microalgae suspension that absorbance is the lowest, P700 signal
Will be the faintest, add frustule and constantly move in suspension, cause signal big ups and downs,
The measurement needs of P700 cannot be met.In the present embodiment microalgae suspension is extracted, its mesh
For test P700 signal；First being filtered on filter membrane by microalgae suspension, microalgae suspends
Concentration after liquid filters is adjusted according to the job demand of staff or tester.After filtration
Microalgae suspension on filter membrane, form stable algae speckle, can according to test P700 signal instrument visit
The size of head diameter, the size of regulation algae speckle sample so that it is more than or equal to probe diameter, in case test
P700 signal is prepared.
Preferably, described preparation method also includes:
Described algae speckle sample is arranged in water white transparency bag by step S30.
Concrete, with reference to shown in Fig. 2, on the basis of a upper embodiment, the algae speckle sample dress that will make
In water white sack, in sack, also include the filter membrane carrying algae speckle.
Preferably, described preparation method also includes:
After described step S30 is provided with the water white transparency bag air-out of algae speckle sample by step S40, carry out close
Envelope processes.
Concrete, with reference to shown in Fig. 2；On the basis of a upper embodiment, for preventing algae speckle sample dehydration,
Affect the test of P700 signal, the sack air-out equipped with algae speckle sample be encapsulated.
Preferably, described preparation method includes:
Described filter membrane also includes: glass fiber filter, or qualitative filter membrane.
Preferably, described preparation method includes:
Described filter membrane diameter is set to 40-50mm, and described filter membrane diameter is more preferably 47mm.
Preferably, described preparation method also includes:
Described step S10 forms algae speckle after membrane filtration, obtains diameter more than 1.5cm algae speckle sample.
Preferably, described preparation method also includes:
Step S10 instills microalgae suspension on filter membrane, for reaching preset concentration, implements the most repeatedly to instill
Described microalgae suspension.
Concrete, when making algae speckle sample, test product used is thin microalgae suspension in early days, due to
The photosynthesis of micro algae growth early stage is higher, but ratio is sparse, dense for reach needed for staff
Degree needs repetitious picked-up, until filter membrane is centrally formed the diameter algae speckle more than 1.5cm.Carry out
During filtration, required filter membrane can use glass fiber filter, or common qualitative filter paper；Simultaneously
The diameter of filter membrane can be chosen to be 40-50mm, for example, 47mm；Demand according to staff is fitted
Adjust to answering property；The diameter of the algae speckle sample of preparation can not be less than the probe diameter of P700 tester, or
The diameter of person's algae speckle sample is more than 1.5cm.
The present invention also provides for the algae speckle Samples EXAMPLE that a kind of preparation method according to aforementioned algae speckle sample prepares.
Concrete, filter membrane instills microalgae suspension, reaches preset concentration；Shape after membrane filtration
Become algae speckle, obtain the diameter algae speckle sample not less than probe diameter；Described algae speckle sample is arranged on nothing
In color transparent bag；After being provided with the water white transparency bag air-out of algae speckle sample, carry out encapsulation process.
When making algae speckle sample, test product used is thin microalgae suspension in early days, due to micro algae growth early stage
Photosynthesis higher, but ratio is sparse, for needed for reaching staff concentration need the most anti-
Multiple picked-up, until filter membrane is centrally formed the diameter algae speckle more than 1.5cm.When filtering, required
The filter membrane wanted can use glass fiber filter, or common qualitative filter paper；The diameter of filter membrane can simultaneously
To be chosen to be 40-50mm, for example, 47mm；Demand according to staff adjusts adaptively；
The diameter of the algae speckle sample of preparation can not be less than the probe diameter of P700 tester, or algae speckle sample
Diameter more than 1.5cm.
The present invention also provides for an embodiment of a kind of method testing P700 signal, with reference to shown in Fig. 3；Bag
Include:
The described test sample of preparation is placed on the test table in P700 measuring instrument by step S10；
Test sample described in step S20 is placed between receiving transducer and the transmitting probe of described test table；
Step S30 records the data message of described receiving transducer.
Concrete, in the present embodiment, the algae speckle sample prepared is applied to measure the signal of P700,
The water white transparency bag that will be equipped with algae speckle sample is put in P700 measuring instrument, and algae speckle sample is placed in P700
Between two probes of measuring instrument, at measurement head alignment algae speckle, carry out P700 measurement；One of them be
Launching the probe of P700 signal, another is the probe receiving P700 signal, and the letter of P700 checked in record
Number state.
Based on above example to the algae speckle sample pair prepared by the preparation method of algae speckle sample and utilization
The test of P700 signal, it is provided that embodiment in greater detail below:
Embodiment one:
By Dual-PAM-100 type P700 measuring instrument, growth early stage thin microalgae suspension sample is surveyed
Amount.With reference to shown in Fig. 4；The P700 signal of thin microalgae suspension sample is low due to frustule concentration, because of
This signal noise is huge, it is impossible to obtain valuable information.
Embodiment two:
By Dual-PAM-100 type P700 measuring instrument, growth dense microalgae suspension sample in latter stage is surveyed
Amount.With reference to shown in Fig. 5；The P700 signal of dense microalgae suspension sample carries due to sample frustule concentration
Height, therefore the noise of P700 has reduced, and the resolution of signal has had certain improvement.But due to its algae
Cell concentration is the most relatively low, adds the frustule motion in suspension, causes signal still to have bigger fluctuation,
High-precision analysis and application cannot be met.
Embodiment three:
Use the method for the invention to strengthen by Dual-PAM-100 type P700 measuring instrument and measure signal: ginseng
Examine shown in Fig. 6；
1. cut-off footpath 47mm glass fiber filter (compared with common qualitative filter paper, glass fiber filter
Optical transmittance is more preferable, is more beneficial for the mensuration of P700 signal), it is placed in Vacuum filtration device.
2. draw microalgae suspension to be measured with pipettor or dropper, instill filter membrane center.For thin microalgae
Suspension, can repeatable operation repeatedly, until filter membrane is centrally formed diameter more than the algae speckle of 1.5cm.
3. the filter membrane with algae speckle being loaded water white plastics valve bag, air-out also seals.In case
Only algae speckle dehydration, affects its P700 activity.
4. the plastics valve bag that will be equipped with algae speckle filter membrane is put in P700 measuring instrument, will measure head alignment algae speckle
Place carries out P700 measurement.
Contrasted further by Fig. 4, Fig. 5, Fig. 6, it is evident that by the method for the present invention, by frustule
Focus on filter membrane surface, form a dense microalgae cell layer, which greatly enhances the density of microalgae cell
And chlorophyll concentration, avoiding microalgae cell movement in suspension simultaneously, P700 signal has obtained significantly
Strengthening, noise fluctuation is the lowest, and signal intensity rule is obvious, and signal has the highest resolution.
Can greatly be strengthened the P700 signal of microalgae suspension by the method for the present invention, reduce signal and make an uproar
Sound, improves signal resolution degree, meets the mensuration requirement to P700 signal of the microalgae sample well.
It should be noted that, above-described embodiment all can independent assortment as required.The above is only this
Bright preferred implementation, it is noted that for those skilled in the art,
Without departing from the principles of the invention, it is also possible to make some improvements and modifications, these improve and profit
Decorations also should be regarded as protection scope of the present invention.
Claims (10)
1. the preparation method of an algae speckle sample, it is characterised in that including:
Step S10 instills microalgae suspension on filter membrane, reaches preset concentration；
Step S10 described in step S20 forms algae speckle after membrane filtration, obtains diameter not less than probe
The algae speckle sample of diameter.
The preparation method of algae speckle sample the most according to claim 1, it is characterised in that also wrap
Include:
Described algae speckle sample is arranged in water white transparency bag by step S30.
The preparation method of algae speckle sample the most according to claim 2, it is characterised in that also wrap
Include:
After described step S30 is provided with the water white transparency bag air-out of algae speckle sample by step S40,
Carry out encapsulation process.
The preparation method of algae speckle sample the most according to claim 1, it is characterised in that including:
Described filter membrane also includes: glass fiber filter, or qualitative filter membrane.
The preparation method of algae speckle sample the most according to claim 1, it is characterised in that including:
Described filter membrane diameter is set to 40-50mm.
The preparation method of algae speckle sample the most according to claim 5, it is characterised in that including:
Described filter membrane diameter is set to 47mm.
7. according to the preparation method of the arbitrary described algae speckle sample of claim 1-6, it is characterised in that
Also include:
Described step S10 forms algae speckle after membrane filtration, obtains diameter more than 1.5cm algae speckle sample
Product.
The preparation method of algae speckle sample the most according to claim 1, it is characterised in that also wrap
Include:
Step S10 instills microalgae suspension on filter membrane, for reaching preset concentration, implements the most anti-
Instill described microalgae suspension again.
9. an algae speckle sample, it is characterised in that: by the preparation as described in claim 1-8 is arbitrary
The algae speckle sample that method prepares.
10. the method testing P700 signal, it is characterised in that application is such as claim 1-8
The algae speckle sample that described preparation method prepares realizes the method for test P700 signal, including:
The described test sample of preparation is placed on the test table in P700 measuring instrument by step S10
On；
Test sample described in step S20 is placed in receiving transducer and the transmitting probe of described test table
Between；
Step S30 records the data message of described receiving transducer.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113607649A (en) * | 2020-09-18 | 2021-11-05 | 中国科学院植物研究所 | Method for improving algae P700 detection signal |
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| CN103396569A (en) * | 2013-08-19 | 2013-11-20 | 南京林业大学 | Bacterial cellulose nanometer optical transparent film preparation method |
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| CN113607649A (en) * | 2020-09-18 | 2021-11-05 | 中国科学院植物研究所 | Method for improving algae P700 detection signal |
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