CN207435435U - A kind of plant protoplast purifies instrument - Google Patents
A kind of plant protoplast purifies instrument Download PDFInfo
- Publication number
- CN207435435U CN207435435U CN201720716589.4U CN201720716589U CN207435435U CN 207435435 U CN207435435 U CN 207435435U CN 201720716589 U CN201720716589 U CN 201720716589U CN 207435435 U CN207435435 U CN 207435435U
- Authority
- CN
- China
- Prior art keywords
- protoplast
- catch tray
- strainer
- electrophoresis
- chloroplaset
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Fee Related
Links
Landscapes
- Apparatus Associated With Microorganisms And Enzymes (AREA)
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
Abstract
The utility model belongs to plant protoplast separating and purifying technology field, specifically disclose a kind of plant protoplast purifying instrument, including tiselius apparatus, sample addition zone, water clock room and Electrophoresis Lab are in turn divided into tiselius apparatus from top to bottom, water clock room is separated with sample addition zone by filter membrane, negative electrode is provided on the left of Electrophoresis Lab, right side is provided with positive electrode, the bottom of Electrophoresis Lab has been respectively communicated with cell fragment catch tray, protoplast catch tray and chloroplaset catch tray from left to right, and strainer is also vertically arranged in Electrophoresis Lab.The method that the plant protoplast purifying instrument method of the utility model utilizes the charged different separated protoplasts by way of electrophoresis of plant protoplast, whole process passes through an electrophoresis, step is simplified, damage and the pollution rate of protoplast is reduced, can sieve in batches.
Description
Technical field
The utility model belongs to plant protoplast separating and purifying technology field, and in particular to a kind of plant cell is former
Raw plastid purifying instrument.
Background technology
Plant protoplast refers to the primary matter group after plant cell wall is removed, and is kept in suitable penetrating fluid
The cell of ball-type.Protoplast occupies an important position in plant cell engineering at present, not still plant cell hybridization
The preferable receptor of good parent or genetically engineered plant is the important materials of RESEARCH ON CELL-BIOLOGY.Plant cell plasm
Body separation is the important technical basis for reaching these purposes.
Traditional protoplast purification process has natural sedimentation, density-gradient centrifugation method.Natural sedimentation will filter and
Sedimentation is combined, and enzymolysis liquid strainer filtering is placed in test tube, and fragment of tissue does not remove parietal cell, protoplast and chloroplaset etc.
Decrease speed is different under the effect of gravity for organelle, and it is plasmic to obtain rule of thumb to draw the more suspension of protoplast
Purifying；Centrifuge floating method be that enzymolysis liquid is placed in centrifuge tube, then by hypertonic solution add to centrifugation bottom of the tube (such as 25% sucrose is molten
Liquid), form an interface between enzymolysis liquid and hypertonic solution, then the horizontal centrifugal by certain speed is not fragment of tissue, complete
The cell body of the full clasmatosis for removing wall is sunken to centrifugation bottom of the tube, intact protoplast concentrate on hypertonic upper strata and enzymolysis liquid it
Between interface on, remained in if organelle such as chloroplaset in enzymolysis liquid, it is careful to draw protoplast liquid layer to be purified.
However traditional natural sedimentation, density-gradient centrifugation method etc. is cumbersome, well damage, yield is low, and passes through density
Gradient centrifugation it is larger to the destruction of protoplast, yield is low.It is necessary to develop the protoplast that a kind of step is simplified, yield is high
Purification devices.
Utility model content
A kind of plant protoplast purifying instrument provided by the utility model solves traditional natural sedimentation, close
Spend that gradient centrifugation etc. is cumbersome, and well damage, yield is low, and by the larger to the destruction of protoplast of density gradient centrifugation,
The problem of yield is low.
The purpose of this utility model is to provide a kind of plant protoplast purifying instrument and purification process, including electrophoresis
Pond is in turn divided into sample addition zone, water clock room and Electrophoresis Lab in the tiselius apparatus from top to bottom, and the water clock room is funnel shaped,
The sample addition zone is connected with the water clock room and water clock room upper end is separated with the sample addition zone by filter membrane, the electricity
Swimming is provided with negative electrode on the left of room, and right side is provided with positive electrode, and the bottom of the Electrophoresis Lab has been respectively communicated with carefully from left to right
Born of the same parents' garbage collection disk, protoplast catch tray and chloroplaset catch tray, the cell fragment catch tray are located at the water clock room
Lower section, the chloroplaset catch tray are set close to the positive electrode, and strainer, the strainer are also vertically arranged in the Electrophoresis Lab
Above the right side wall of protoplast catch tray.
Preferably, in plant protoplast purifying instrument, the aperture φ of the filter membrane is 50-100 microns.
Preferably, in plant protoplast purifying instrument, the filter membrane is shelved at the upper end port of water clock room.
Preferably, in plant protoplast purifying instrument, the aperture φ of the strainer is 10 microns.
Preferably, in plant protoplast purifying instrument, the upper end of the Electrophoresis Lab is close to the positive electrode
Place offers strainer jack, and offers strainer sliding slot, the width of the strainer on two opposite side walls of Electrophoresis Lab respectively
For degree less than the length of the strainer jack, the thickness of the strainer is less than the width of the strainer jack, and the strainer
Both sides are slidably connected with two strainer sliding slot one-to-one corresponding.
Preferably, in plant protoplast purifying instrument, the outside of the tiselius apparatus is additionally provided with direct current
Swimming instrument, and the cathode of the direct current electrophoresis apparatus is electrically connected with the negative electrode, and anode is electrically connected with the positive electrode.
Preferably, in plant protoplast purifying instrument, the cell fragment catch tray, the protoplast
Catch tray and the chloroplaset catch tray are removably attachable to the bottom of the Electrophoresis Lab, and concrete structure is：The electrophoresis
The bottom of room has been respectively communicated with cell fragment runner pipe from left to right, protoplast receives runner pipe and chloroplaset runner pipe, described
The openend of cell fragment catch tray is spirally connected with cell fragment circulation bottom of the tube, the openend of the protoplast catch tray
Circulation bottom of the tube is received with the protoplast to be spirally connected, openend and the chloroplaset circulation bottom of the tube of the chloroplaset catch tray
It is spirally connected.
Preferably, in plant protoplast purifying instrument, the cell fragment catch tray and the cell are broken
Between piece runner pipe, the protoplast catch tray and the protoplast receive runner pipe between, the chloroplaset catch tray with
Sealing ring is both provided between the chloroplaset runner pipe.
Compared with prior art, a kind of plant protoplast purifying instrument and purification process provided by the utility model,
It has the advantages that：
1st, the plant protoplast purifying instrument of the utility model passes through electrophoresis using the charged difference of plant protoplast
Mode separated protoplast method, whole process only needs a kind of buffer solution, simplifies step, reduces protoplast
Damage and pollution rate, can sieve in batches.Solve in the prior art natural sedimentation, density-gradient centrifugation method and etc. it is multiple
The problem of miscellaneous, yield is low, and by the larger to the destruction of protoplast of density gradient centrifugation, and yield is low.
Description of the drawings
Fig. 1 is the structure diagram that the utility model plant protoplast purifies instrument；
Fig. 2 is the fundamental diagram that the utility model plant protoplast purifies instrument.
Reference sign：1. sample addition zone, 11. filter membranes, 2. water clock rooms, 3. Electrophoresis Labs, 31. negative electrodes, 32. positive electrodes,
4. cell fragment catch tray, 5. protoplast catch trays, 6. chloroplaset catch trays, 33. strainers, 331. strainer sliding slots, 7. direct currents
Electrophoresis apparatus.
Specific embodiment
The utility model is described in detail in the following with reference to the drawings and specific embodiments, but it is new to should not be construed as this practicality
The limitation of type.The test method of actual conditions is not specified in the following example, is usually operated according to normal condition.When embodiment is given
When going out numberical range, it should be appreciated that unless the utility model is otherwise noted, two endpoints and two endpoints of each numberical range
Between any one numerical value can be selected.Unless otherwise defined, all technical and scientific terms for being used in the utility model with
The normally understood meaning of those skilled in the art of the present technique is identical.
The utility model provides a kind of plant protoplast purifying instrument, specifically as shown in Figure 1, including by insulating materials
The tiselius apparatus being prepared is in turn divided into sample addition zone 1, water clock room 2 and Electrophoresis Lab 3 in the tiselius apparatus from top to bottom, described
Water clock room 2 is funnel shaped, and sample addition zone 1 is connected with water clock room 2 and 2 upper end port of water clock room and the sample addition zone 1 are logical
Filter membrane 11 separates, and the aperture φ of the filter membrane 11 is 50-100 microns, and the left side of the Electrophoresis Lab 3 is provided with negative electrode 31,
Right side is provided with positive electrode 32, and the bottom of the Electrophoresis Lab 3 has been respectively communicated with cell fragment catch tray 4, plasm from left to right
Body catch tray 5 and chloroplaset catch tray 6, between cell fragment catch tray 4, protoplast catch tray 5 and chloroplaset catch tray 6
Spacing is 2-5cm, and the cell fragment catch tray 4 is located at the underface of the water clock room 2, and 4 He of cell fragment catch tray
The width of water clock room 2, equal length, width, length are 2-8cm, width, the length 2- of the protoplast catch tray 5
8cm, protoplast catch tray 5 are located at the right side of the cell fragment catch tray 4, and the chloroplaset catch tray 6 is located at plasm
It the right side of body catch tray 5 and sets close to the positive electrode 32, between the chloroplaset catch tray 6 and the positive electrode 32
Distance for 0-1cm, dismountable strainer 33 is also vertically arranged in the Electrophoresis Lab 3, the aperture φ of the strainer 33 is 10
Micron, the strainer 33 are located above the right side wall of protoplast catch tray 5.
It should be noted that the outside of the tiselius apparatus is additionally provided with direct current electrophoresis apparatus 7 (enough from 61 instrument plants), and
The cathode of the direct current electrophoresis apparatus 7 is electrically connected with the negative electrode 31, and anode is electrically connected with the positive electrode 32, to realize electrophoresis
The electrophoresis functions in pond.
It should be noted that the dismounting and cleaning of device for convenience, cell fragment catch tray 4, protoplast catch tray 5
The bottom of the Electrophoresis Lab 3 is removably attachable to chloroplaset catch tray 6, concrete structure is：The bottom of the Electrophoresis Lab 3
Cell fragment runner pipe, protoplast receipts runner pipe and chloroplaset runner pipe have been respectively communicated with, the cell fragment catch tray 4
Openend is spirally connected with cell fragment circulation bottom of the tube, openend and the protoplast of the protoplast catch tray 5
It receives circulation bottom of the tube to be spirally connected, openend and the chloroplaset circulation bottom of the tube of the chloroplaset catch tray 6 are spirally connected.
Between the cell fragment catch tray 4 and the cell fragment runner pipe, the protoplast catch tray 5 with it is described
Between protoplast receives runner pipe, sealing ring is both provided between the chloroplaset catch tray 6 and the chloroplaset runner pipe, is used
In preventing leakage.
It should be noted that the filter membrane 11 is shelved at 2 upper end port of water clock room, to realize that filter membrane 11 removably connects
It is connected on water clock room 2, is conveniently replaceable filter membrane 11.
It should be noted that the upper end of the Electrophoresis Lab 3 offers strainer jack at the positive electrode 32, strainer is inserted
Hole and the distance of chloroplaset catch tray 6 are 0-0.5cm, and offer strainer respectively on two opposite side walls of Electrophoresis Lab 3
Sliding slot 331, the width of the strainer 33 are less than the length of the strainer jack, and the thickness of the strainer 33 is inserted less than the strainer
The width in hole, and the both sides of the strainer 33 are slidably connected with two one-to-one corresponding of strainer sliding slot 331, and strainer 33 is enable to pass through
It the strainer jack and is skidded off along two strainer sliding slots 331, is conveniently replaceable strainer 33.
The plant protoplast of the utility model purifies instrument use principle as shown in Fig. 2, being：Protoplast surface band
There is negative electrical charge (- 10-30mV), parietal cell is not gone using protoplast, chloroplaset, fragment of tissue and completely to have the suspension of electric field
Charge is different in solution, shape is different, gravity formation of different sizes lateral drift speed is different realizes the purpose isolated and purified.
Electrophoresis Lab 3 is put into electrophoresis liquid, and the cathode of the direct current electrophoresis apparatus 7 is electrically connected with the negative electrode 31, anode and institute
It states positive electrode 32 to be electrically connected, the protoplast mixture suspension prepared the methods of enzymolysis is being loaded by certain speed dropwise addition
In room 1, protoplast mixture suspension, into water clock room 2, is then gradually dropped Electrophoresis Lab 3 by filter membrane 11 through water clock room 2
Interior, plant protoplast in Electrophoresis Lab 3 is moved to 32 direction of positive electrode, is stagnated on the strainer 33 before positive electrode 32, electrophoresis
Protoplast sedimentation falls into protoplast catch tray 5 after stopping, protoplast catch tray 5 is taken out the plasm for obtaining purifying
Body, and the cell of cell fragment and non-broken wall then falls into cell fragment catch tray 4 due to gravity；Chloroplaset then penetrates strainer
33 fall into chloroplaset catch tray 6.
This plant protoplast purifies instrument, and overcoming to have showed has a series of problems existing for technology, former using plant
The method of the raw charged different separated protoplasts by way of electrophoresis of plastid, whole process only need a kind of buffer solution, simplify
Step, reduces damage and the pollution rate of protoplast, can sieve in batches.
A kind of application method using above-mentioned plant protoplast purifying instrument is provided below to comprise the following steps：
S1 fills the mannitol solution of 0.5mol/L in Electrophoresis Lab 3, as buffer solution, opens power supply, adjusts direct current electrophoresis
Instrument makes the electric field strength of direct current electrophoresis apparatus adjust in 1.5-5v/cm, carries out electrophoresis；
S2, after electrophoresis starts, by the 10mL plant protoplast mixture suspension prepared according to 6-12 drops/minute
Speed is added dropwise in sample addition zone 1 up to being added dropwise, and then proceedes to electrophoresis 30min；
S3 closes power supply, collects the protoplast in protoplast catch tray 5.
Although the preferred embodiment of the utility model has been described, those skilled in the art once know substantially
Creative concept can then make these embodiments other change and modification.So appended claims are intended to be construed to wrap
It includes preferred embodiment and falls into all change and modification of the scope of the utility model.
Obviously, those skilled in the art can carry out the utility model various modification and variations without departing from this practicality
New spirit and scope.If in this way, these modifications and variations of the utility model belong to the utility model claims and
Within the scope of its equivalent technologies, then the utility model is also intended to comprising including these modification and variations.
Claims (8)
1. a kind of plant protoplast purifies instrument, which is characterized in that including tiselius apparatus, in the tiselius apparatus from top to bottom according to
Secondary to be divided into sample addition zone (1), water clock room (2) and Electrophoresis Lab (3), the water clock room (2) is funnel shaped, the sample addition zone (1)
It is connected with the water clock room (2) and water clock room (2) upper end is separated with the sample addition zone (1) by filter membrane (11), it is described
It is provided with negative electrode (31) on the left of Electrophoresis Lab (3), right side is provided with positive electrode (32), and the bottom of the Electrophoresis Lab (3) is from a left side
It is broken that cell fragment catch tray (4), protoplast catch tray (5) and chloroplaset catch tray (6), the cell have been respectively communicated with to the right side
Piece catch tray (4) is located at the lower section of the water clock room (2), and the chloroplaset catch tray (6) is set close to the positive electrode (32),
Strainer (33) is also vertically arranged in the Electrophoresis Lab (3), the strainer (33) is located at the right side wall of protoplast catch tray (5)
Top.
2. plant protoplast according to claim 1 purifies instrument, which is characterized in that the aperture of the filter membrane (11)
φ is 50-100 microns.
3. plant protoplast according to claim 1 purifies instrument, which is characterized in that the filter membrane (11) is shelved on
At water clock room (2) upper end port.
4. plant protoplast according to claim 1 purifies instrument, which is characterized in that the aperture of the strainer (33)
φ is 10 microns.
5. plant protoplast according to claim 1 purifies instrument, which is characterized in that the Electrophoresis Lab (3) it is upper
End offers strainer jack at the positive electrode (32), and is opened up respectively on two opposite side walls of Electrophoresis Lab (3)
There is strainer sliding slot (331), the width of the strainer (33) is less than the length of the strainer jack, and the thickness of the strainer (33) is small
In the width of the strainer jack, and the both sides of the strainer (33) correspond the company of slip with two strainer sliding slots (331)
It connects.
6. plant protoplast according to claim 1 purifies instrument, which is characterized in that the outside of the tiselius apparatus is also
Be provided with direct current electrophoresis apparatus (7), and the cathode of the direct current electrophoresis apparatus (7) is electrically connected with the negative electrode (31), anode with
Positive electrode (32) electrical connection.
7. plant protoplast according to claim 1 purifies instrument, which is characterized in that the cell fragment catch tray
(4), the protoplast catch tray (5) and the chloroplaset catch tray (6) are removably attachable to the Electrophoresis Lab (3)
Bottom, concrete structure are：The bottom of the Electrophoresis Lab (3) has been respectively communicated with cell fragment runner pipe, protoplast from left to right
Receive runner pipe and chloroplaset runner pipe, openend and the cell fragment circulation bottom of the tube of the cell fragment catch tray (4)
It is spirally connected, openend and the protoplast of the protoplast catch tray (5) receive circulation bottom of the tube and be spirally connected, and the chloroplaset is received
The openend of catch basin (6) is spirally connected with chloroplaset circulation bottom of the tube.
8. plant protoplast according to claim 7 purifies instrument, which is characterized in that the cell fragment catch tray
(4) between the cell fragment runner pipe, between the protoplast catch tray (5) and the protoplast receipts runner pipe,
Sealing ring is both provided between the chloroplaset catch tray (6) and the chloroplaset runner pipe.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201720716589.4U CN207435435U (en) | 2017-06-20 | 2017-06-20 | A kind of plant protoplast purifies instrument |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201720716589.4U CN207435435U (en) | 2017-06-20 | 2017-06-20 | A kind of plant protoplast purifies instrument |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN207435435U true CN207435435U (en) | 2018-06-01 |
Family
ID=62396021
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201720716589.4U Expired - Fee Related CN207435435U (en) | 2017-06-20 | 2017-06-20 | A kind of plant protoplast purifies instrument |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN207435435U (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107058075A (en) * | 2017-06-20 | 2017-08-18 | 商丘师范学院 | A kind of plant protoplast purifying instrument and purification process |
-
2017
- 2017-06-20 CN CN201720716589.4U patent/CN207435435U/en not_active Expired - Fee Related
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107058075A (en) * | 2017-06-20 | 2017-08-18 | 商丘师范学院 | A kind of plant protoplast purifying instrument and purification process |
| CN107058075B (en) * | 2017-06-20 | 2023-10-20 | 商丘师范学院 | Plant cell protoplast purification instrument and purification method |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN108587902A (en) | Cell sorting devices based on dielectrophoresis and its screening technique | |
| CN103466760B (en) | Electromagnetic type mud-water separation tower and electromagnetic type mud-water separation method | |
| JP2015503929A5 (en) | ||
| CN101648086A (en) | Rapid precipitating method and rapid precipitating and separating device | |
| CN207435435U (en) | A kind of plant protoplast purifies instrument | |
| CN202482320U (en) | Cell filter | |
| CN103756893A (en) | Foam concentration and harvest device for microalgae | |
| KR101726286B1 (en) | Collecting apparatus for extracellular vesicles and method for using the same | |
| CN202397870U (en) | Blood separating device | |
| CN203291633U (en) | Solid-liquid separating device for waste water suspended solids | |
| CN110684655B (en) | Microalgae separation gradient magnetic stabilization fluidized bed device and microalgae harvesting method thereof | |
| CN107058075A (en) | A kind of plant protoplast purifying instrument and purification process | |
| CN205313547U (en) | Centrifuging tube suitable for a single karyocyte layering | |
| CN205188276U (en) | Tumor cells and lymphocyte separator | |
| CN214654995U (en) | Cell autosegregation instrument | |
| CN212955136U (en) | Device for quickly separating exosomes | |
| CN209338506U (en) | A kind of novel microalgae enrichment facility | |
| CN113567198A (en) | Extraction device and extraction method for micro-plastic in soil/sediment | |
| CN206273314U (en) | A kind of Chinese medicine alcohol precipitation or the heavy dregs of a decoction separating liquid medicine device of water | |
| CN203513341U (en) | Electromagnetic mud-water separation column | |
| CN203389374U (en) | Dilute sulfuric acid mud tank | |
| CN206910527U (en) | Original plant juice extraction element based on protect liver | |
| CN207738497U (en) | Zwitterion separator in a kind of water | |
| CN204474396U (en) | A kind of ceramic membrane filter device | |
| CN205473754U (en) | Cell filter |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20180601 Termination date: 20210620 |
|
| CF01 | Termination of patent right due to non-payment of annual fee |