CN206114666U - Be used for box -packed putting of tumor cells nucleic acid aptamer screening card - Google Patents
Be used for box -packed putting of tumor cells nucleic acid aptamer screening card Download PDFInfo
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- CN206114666U CN206114666U CN201621122587.4U CN201621122587U CN206114666U CN 206114666 U CN206114666 U CN 206114666U CN 201621122587 U CN201621122587 U CN 201621122587U CN 206114666 U CN206114666 U CN 206114666U
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- culture dish
- nucleic acid
- cartridge
- tissue culture
- rifle
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Abstract
The utility model discloses a be used for box -packed putting of tumor cells nucleic acid aptamer screening card, including card box main part, cell culture dish adapter device, reagent strorage device. A card box main part includes mainly that spear tip, pivot, clamp plate and gas circuit install the filter membrane at calorie lateral wall of box main part. The cell culture dish adapter comprises the semi -closed round containers of one, shaft and fixed plate, and the fixed plate comes fixed culture dish adapter through the airtight joint with calorie box main part. Reagent strorage device comprises the pore volume ware of a plurality of salients, with calorie airtight combination of box main part for deposit the reagent used etc. In the screening experiment, two hole sites of edge are porose needles and passageway respectively, is used for carrying on the liquid circulation between two box bodys. One round or many rounds of screenings of tumor cells nucleic acid aptamer can be realized to this card box, have avoided cross contamination's problem, and the accessible self -reacting device is realized simultaneously, improves the efficiency of nucleic acid aptamer screening greatly.
Description
Technical field
This utility model is related to a kind of for tumor cell nucleic acid aptamers screening cartridge device, it is adaptable to tumor cell
The research experiment screened by aptamer, simultaneously can be used for other species aptamer screening tests, such as albumen, outer
Secrete body etc..
Background technology
Aptamer is the oligonucleotide sequence that a bit of Jing in-vitro screenings are obtained, and can carry out high parent with corresponding part
With the combination of power and strong specificity, it appear as chemical-biological educational circles and biomedical boundary provide it is a kind of new efficient quick
The research platform of identification, and good application prospect is illustrated in many aspects.
At present, carry out the screening of tumor cell nucleic acid aptamers main by having been manually done, inefficiency is fitted from screening nucleic acid
Ligand screening starts to being fully completed, and needs to carry out 20 wheel left and right experiments, and whole process needs half a year to 1 year, wherein most heavy
The problem wanted is that cross-contamination occurs, is mainly derived from PCR, aptamers transfer etc., once there is pollution, this wheel or even whole
Individual the selection result just occurs error, it is necessary to re-start screening.At present, it is to avoid the method for cross-contamination mainly passes through will be every
Individual step is divided among different experiments room and is operated, and laboratory is aerated, is sterilized, and this mode needs to take larger ground
Side's arrangement laboratory, and it is higher to laboratory technician's requirement, it is even if so, the aerial nucleic acid that is scattered times cannot so be eliminated, real
In the experiment on border, cross-contamination issue is present always.For the problems referred to above, the utility model proposes a kind of closed, assembling letter
Cartridge device that is single, can be used for automation mechanized operation, after reagent is prepared, all operations are carried out within the enclosed space, aptamers
The passage being displaced through between cartridge is completed, and eliminates polluter, can complete a wheel or multi-turns screen process of aptamer.
The content of the invention
This utility model purpose is to provide a kind of tumor cell nucleic acid aptamers screening cartridge device, will be liquid relief, PCR anti-
Should, aptamers transfer etc. operation one closing cartridge in carry out, it is to avoid cross-contamination issue, and be capable of achieving automatization behaviour
Make.
It is according to a technical scheme of the present utility model, a kind of to screen cartridge device for tumor cell nucleic acid aptamers,
Including housing, mainly it is made up of three parts in housing, including cartridge bodies, Tissue Culture Dish adapter apparatus, reagent storage
Device；Tissue Culture Dish adapter apparatus and reagent storage device are set up in parallel under cartridge bodies；Described cartridge bodies are main
Including for pipette reagent rifle point, control rifle point horizontal anomalous movement rotating shaft, control rifle point vertical movement pressing plate and be used for
Control rifle point draws the gas circuit with discharging liquid, and its center platen lower end is connected with rifle ogival, and pressing plate upper end is projected to outside housing
Portion is connected with pressing plate threaded shaft；Pressing plate lower end is connected with rifle ogival, and pressing plate threaded shaft realizes the vertical movement of pressing plate；Gas
Road one end is connected with rifle point upper side position, and one end connects external path by case top；Rotating shaft is connected with rifle point, and one end projects
Outside housing；Air filter film is installed in the side wall of housing, for air pressure balance in keeping box；Tissue Culture Dish adapter apparatus
It is made up of Tissue Culture Dish adapter, Tissue Culture Dish adapter holder, Tissue Culture Dish adapter holder includes culture
Ware rotating shaft and fixed plate, culture dish rotating shaft are connected with Tissue Culture Dish adapter, and one end is projected to outside cartridge bodies；Fixed plate
With the closed combination of cartridge bodies；Described reagent storage device and the closed combination of cartridge bodies, described reagent storage device
It is made up of several downward projection of pore volume devices, is provided with two hole positions and is respectively equipped with hole pin and passage.
Described air filter film aperture is less than aptamers.
Described cartridge bodies have one or more rifle point, rotating shaft, pressing plates.
The Tissue Culture Dish adapter can be changed according to Tissue Culture Dish size and shape, and can carry out a left side
Right flip-flop movement.
With cartridge bodies to be detachably connected, described hole container bottom is hemispherical or cone to the reagent storage device
Shape, described pore volume device are a row or multi-row.
The hole pin is slender pipeline, penetrates the closed hole position of rubber closure, and passage is connected with the side wall of a pore volume device.
The generation of air pressure in the air flue of described cartridge bodies is made an excuse external air pump by surface and is realized.
Beneficial effect
All carried out in closed cartridge using the whole aptamer screening process of device of the present utility model, wherein holding
The PCR reactions for being also easy to produce cross-contamination occur in cartridge, and library transfer is carried out in special airtight passage, it is ensured that amplification
Product will not be spilt in air, it is to avoid the problem of cross-contamination, while carry out aptamer screening by cartridge can lead to
Self-reacting device realization is crossed, this will greatly improve the efficiency of aptamer screening.
Description of the drawings
Fig. 1 cartridge overall appearance figures
Fig. 2 cartridge bodies outside drawings；
Fig. 3 cartridge bodies cut-away views；
Fig. 4 cartridge Tissue Culture Dish adapter apparatus outside drawings；
Fig. 5 cartridge reagent storage device appearance figures；
Fig. 6 cartridges entirety front section view；
Fig. 7 cartridges entirety right side sectional view；
Wherein：1 cartridge bodies, 2 Tissue Culture Dish adapter apparatus, 3 reagent storage devices, 31 air-path interfaces, 32 pressing plates
33 pressing plate threaded shafts, 34 rifles point, 35 springs, 36 rifles point adapter, 37 rotating shafts, 38 Tissue Culture Dish adapters, the training of 39 cells
Foster ware adapter holder, 40 cleanout fluid storage tanks, 41 pancreatin Digestive system storage tanks, 42TE buffer storage tanks, 43PCR reactions
Groove, 44 aptamers transfer storage tank, 45 holes point, 46 passages, 47 library high temperature storage tanks, 48 library low-temp storage grooves, 49 combine
Liquid storage tank, 50 waste liquid storage tanks, 51 air filter films
Specific embodiment
Specific embodiment is screened with reference to HepG2 nucleus aptamers and accompanying drawing does to this utility model and more enters one
Explain on step ground.Embodiment is merely to illustrate this utility model, but is not used to limit practical range of the present utility model.
It is a kind of to screen cartridge device for tumor cell nucleic acid aptamers, mainly it is made up of three parts, including cartridge master
Body, Tissue Culture Dish adapter apparatus, reagent storage device.Cartridge bodies are mainly included for pipetting the rifle point of reagent, controlling
Pressing plate and draw the gas with discharging liquid for controlling rifle point that the rotating shaft of rifle point horizontal anomalous movement, control paracone move vertically
Road.Wherein rotating shaft one end is projected to outside cartridge, and head is in-line or other shapes interface, is stirred by hand or motor connection
Interface is rotated, and controls pivot, and rifle point is connected to rotating shaft, the point horizontal movement of rotating shaft rotational movement rifle by adapter；
Pressing plate lower end is connected with rifle ogival, and upper end is a kind of female thread structure, is turned with the screw thread for being fixed on cartridge surface by screw thread
Axle connects, and the thread spindle other end is projected outside cartridge, and head is in-line or other shapes interface, stirs by hand or motor connects
Connection interface is rotated, control threaded shaft motion, and band dynamic pressure plate vertical movement, when pressing plate is moved downward, oppresses rifle ogival, rifle
Point is moved downward, while the spring between rifle point and rifle point adapter is compressed；When pressing plate is moved upwards, rifle point is adapted to rifle point
Spring between device recovers, and rifle point is moved upwards.Gas circuit mainly includes that rifle point side position interface, cartridge top connection external path connect
Mouth and pipeline connecting the two, the access interface outside cartridge top connection can connect the pressure generators such as air pump, internal
Air filter film is fixed with, filter sizes are less than aptamers, prevent aptamers from spilling in pressure generator, produce pollution, together
When be fixed with filter membrane in the side wall of cartridge bodies, filter sizes are less than aptamers, put down with external pressure for air pressure in keeping box
Weighing apparatus.Tissue Culture Dish adapter is made up of a semiclosed bulge, Tissue Culture Dish adapter holder, described cell
Culture dish fixture includes rotating shaft and fixed plate.Bulge is used for storage immobilized-cell culture ware, with training inside container
Directly contact outside foster ware, fixes culture dish by frictional force or other devices；Rotating shaft is seamlessly connected with bulge, and one section is dashed forward
Go out to cartridge, head is in-line or other shapes interface, stir by hand or motor connection interface is rotated, control rotating shaft
Motion, swings so as to control adapter, for controlling liquid aggregation position in culture dish；Fixed plate by with cartridge bodies
Tight engagement fixing culture dish adapter.Reagent storage device is made up of the pore volume device of multiple protrusions, for depositing experiment
In the reagent used, with the closed combination of cartridge bodies, on side, two hole positions have hole pin and passage, hole needle section and serum cap respectively
It is in close contact, the entrance of passage sealed by rubber stopper, the hole needle section rubber stopper of cartridge and the rubber of another cartridge closed channel
After plug is combined closely, acupuncture broken rubber stopper in external force pressure hole enters passage, realizes two cartridge hole position aptamers transfers, completes
Afterwards, pulled-out holes pin.
Embodiment
HepG2 cells carry out wheel aptamer screening key step has library process, DNA to be combined with cell, cell
Separate and the amplification of broken, aptamer, aptamer transfer.Used reagent has single-stranded DNA banks, with reference to liquid, clearly
Washing liquid, pancreatin Digestive system, TE buffer, asymmetric PCR reaction reagent etc., this utility model substantially have 8 to hold reagent
Chamber and a waste liquid slot chamber.
This utility model is integrally a kind of square cartridge, and cartridge is made up of three parts：Cartridge bodies 1, cell culture
Ware adapter apparatus 2 and reagent storage device 3, three is closely entrenched togather.There is air-path interface 31 at the top of cartridge bodies 1, use
In connection air pump, cartridge inner tip has pressing plate 32, pressing plate threaded shaft 33, and threaded shaft 33 is connected with pressing plate 32, rotatably
Threaded shaft 33 controls the vertical movement of pressing plate 32, engages with pressing plate 32 at the top of rifle point 34, and rifle 34 cavitys of point pass through 35 He of spring
Rifle point adapter 36, and fixed by rifle point adapter 36, pressing plate 32 is moved downward, and compressing rifle point 34 is moved downward, spring 35
Compression, when pressing plate 32 is moved upwards, spring 35 recovers, and rifle point 34 is moved upwards, and rifle point adapter 36 passes through screw thread with rotating shaft 37
Connection, rotating shaft 37 rotarily drive rifle point 36 horizontal movement of adapter, so as to control rifle 34 horizontal movements of point, 51 aperture of air filter film
Less than aptamers size, aptamers cannot pass through.Tissue Culture Dish adapter apparatus 2 are by Tissue Culture Dish adapter 38 and cell
Culture dish adapter holder 39 is constituted, and Tissue Culture Dish adapter holder 39 carries out fluid-tight engagement with 1 lower end of cartridge bodies.
Reagent storage device 3 is total with regard to 8 reagent storage grooves, and respectively cleanout fluid storage tank 40, pancreatin Digestive system storage tank 41, TE delays
Rush liquid storage tank 42, PCR reactive tanks 43, aptamers transfer storage tank 44, library high temperature storage tank 47, library low-temp storage groove
48th, with reference to 49,1 waste liquid tank 50 of liquid storage tank, its mesopore point 45 is connected with aptamers transfer storage tank 44, outlet rubber closure
Seal, passage 46 is connected with library high temperature storage tank 47.Carry out the first round aptamer screening start before, DNA library is put
Enter library high temperature storage tank 47, be put into reference to liquid and PCR reactive tanks 43, TE buffering be put into reference to liquid storage tank 49, PCR reaction reagents
Liquid be put into TE buffer storage tank 42, pancreatin Digestive system be put into pancreatin Digestive system storage tank 41, cleanout fluid be put into cleanout fluid storage
Tissue Culture Dish is put into Tissue Culture Dish adapter 38 by groove 40, after completing, cartridge bodies 1 is adapted to Tissue Culture Dish
Device device 2, reagent storage device 3 are completed.Library high temperature storage tank 47 is put into after 95 DEG C of metal bath 5min, rotating shaft
37, rifle point 34 is moved to directly over library high temperature storage tank 47, pressing plate threaded shaft 33 is rotated so that pressing plate 32 is moved down, by rifle
Point 34 is depressed into inside library high temperature storage tank 47, and library is sucked sharp 34 chambers of rifle by plunger displacement pump be connected with air-path interface 31
Body, rotates pressing plate threaded shaft 33, moves on pressing plate 32, is acted on by 35 elastic force of spring, and rifle point 34 moves to library high temperature storage tank
Above in the of 47, rotating shaft 37, rifle point 34 are moved to directly over library low-temp storage groove 48, are rotated pressing plate threaded shaft 33 and are pushed pressing plate
32, rifle point 34 is moved to inside library low-temp storage groove 48, rifle 34 cavity Chinese libraries of point is put into library low temperature by plunger displacement pump and is deposited
Put in groove 48, after 0 DEG C of storage 10min, with sharp 34 absorption library low-temp storage groove, 48 Chinese library of rifle to liquid storage tank 49 is combined, lead to
Cross after 34 piping and druming of rifle point mixed with reference to liquid in liquid storage tank 49, with rifle point 34 draw cleanout fluid in cleanout fluid storage tank 40 put to
In Tissue Culture Dish adapter 38 in Tissue Culture Dish, gently with rifle point after 38 2min of left-right rotation Tissue Culture Dish adapter
34 draw Tissue Culture Dish supernatants, put to waste liquid storage tank 50, after repeated washing 3 times, are drawn with reference to liquid storage tank with rifle point 34
In 49, liquid is moved in Tissue Culture Dish adapter 38 in Tissue Culture Dish, the gently left-right rotation cell culture in 4 DEG C of environment
38 2h of ware adapter, draws Tissue Culture Dish aqueous supernatant with rifle point 34, puts to waste liquid storage tank 50, is drawn with rifle point 34 clear
In washing liquid storage tank 40, cleanout fluid is put into Tissue Culture Dish adapter 38 in Tissue Culture Dish, gently left-right rotation cell culture
Tissue Culture Dish supernatant is drawn with rifle point 34 after 38 2min of ware adapter, is put to waste liquid storage tank 50, after repeated washing 3 times, inhaled
In taking TE buffer storage tank 42, TE buffer is into Tissue Culture Dish, then drawn in pancreatin Digestive system storage tank 41 with rifle point 34
Pancreatin Digestive system is moved to liquid in Tissue Culture Dish in PCR reactive tanks 43 with rifle point 34 to Tissue Culture Dish 10min, is mixed
Amplified production, after amplification terminates, is moved to aptamers transfer storage tank 44 with rifle point 34, by hole point 45 by even laggard performing PCR amplification
The passage 46 that another aptamer put reagent (except library) well and be completed screens cartridge is directed at, is gently used
Hole point 45 is pierced into passage 46 by power, in aptamers transfer groove 44 pcr amplification product flow in another cartridge by hole point 45 and
In the library high temperature storage tank 47 of the connection of passage 46, transfer is finished and is then abandoned first cartridge, so far, first round nucleic acid adaptation
Body screening is completed, and directly can carry out the second wheel or the n-th wheel experiment by above-mentioned steps, until aptamer screening is completed.
Whole aptamer screening process is all carried out in closed cartridge, wherein easily producing the PCR of cross-contamination
Reaction occurs in cartridge, and library transfer is carried out in special airtight passage, it is ensured that amplified production will not spill into air
In, it is to avoid the problem of cross-contamination, while aptamer screening is carried out by cartridge can be realized by self-reacting device, this
The efficiency of aptamer screening will be greatly improved.
Claims (7)
1. it is a kind of to screen cartridge device for tumor cell nucleic acid aptamers, it is characterised in that including housing, mainly to have in housing
Three parts constitute, including cartridge bodies（1）, Tissue Culture Dish adapter apparatus（2）, reagent storage device（3）；Cartridge bodies
（1）Under be set up in parallel Tissue Culture Dish adapter apparatus（2）With reagent storage device（3）；Described cartridge bodies mainly include
For pipetting the rifle point of reagent（34）, control rifle point（34）The rotating shaft of horizontal anomalous movement（37）, control rifle point（34）Vertical movement
Pressing plate（32）And for controlling rifle point（34）Draw the gas circuit with discharging liquid, its center platen（32）Lower end is connected with rifle ogival
Connect, pressing plate（32）Upper end projects to hull outside and pressing plate threaded shaft（33）Connection；Pressing plate（32）Lower end is connected with rifle ogival
Connect, pressing plate threaded shaft（33）Realize the vertical movement of pressing plate；Gas circuit one end is connected with rifle point upper side position, and one end passes through housing
Top connection external path；Rotating shaft is connected with rifle point, and one end protrusion is outside housing；Air filter film is installed in the side wall of housing,
For air pressure balance in keeping box；Tissue Culture Dish adapter apparatus（2）By Tissue Culture Dish adapter（38）, Tissue Culture Dish
Adapter holder（39）Composition, Tissue Culture Dish adapter holder（39）Including culture dish rotating shaft and fixed plate, culture dish
Rotating shaft is connected with Tissue Culture Dish adapter, and one end is projected to outside cartridge bodies；The closed combination of fixed plate and cartridge bodies；
Described reagent storage device and the closed combination of cartridge bodies, described reagent storage device（3）It is downward projection of by several
Pore volume device is constituted, and is provided with two hole positions and is respectively equipped with hole pin and passage.
2. it is according to claim 1 to screen cartridge device for tumor cell nucleic acid aptamers, it is characterised in that described
Air filter film aperture is less than aptamers.
3. it is according to claim 1 to screen cartridge device for tumor cell nucleic acid aptamers, it is characterised in that described
Cartridge bodies have one or more rifle points（34）, rotating shaft（37）, pressing plate（32）.
4. it is according to claim 1 to screen cartridge device for tumor cell nucleic acid aptamers, it is characterised in that described thin
Born of the same parents' culture dish adapter can be changed according to Tissue Culture Dish size and shape, and can carry out left and right flip-flop movement.
5. it is according to claim 1 to screen cartridge device for tumor cell nucleic acid aptamers, it is characterised in that the examination
With cartridge bodies to be detachably connected, described hole container bottom is hemispherical or taper, described pore volume device to agent storing unit
For a row or multi-row.
6. it is according to claim 1 to screen cartridge device for tumor cell nucleic acid aptamers, it is characterised in that the hole
Pin is slender pipeline, penetrates the closed hole position of rubber closure, and passage is connected with the side wall of a pore volume device.
7. it is according to claim 1 to screen cartridge device for tumor cell nucleic acid aptamers, it is characterised in that described
The generation of air pressure in the air flue of cartridge bodies is made an excuse external air pump by surface and is realized.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201621122587.4U CN206114666U (en) | 2016-10-13 | 2016-10-13 | Be used for box -packed putting of tumor cells nucleic acid aptamer screening card |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201621122587.4U CN206114666U (en) | 2016-10-13 | 2016-10-13 | Be used for box -packed putting of tumor cells nucleic acid aptamer screening card |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN206114666U true CN206114666U (en) | 2017-04-19 |
Family
ID=58529454
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201621122587.4U Withdrawn - After Issue CN206114666U (en) | 2016-10-13 | 2016-10-13 | Be used for box -packed putting of tumor cells nucleic acid aptamer screening card |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN206114666U (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106353492A (en) * | 2016-10-13 | 2017-01-25 | 东南大学 | Cassette device for screening tumor cell aptamers |
| CN108004324A (en) * | 2017-12-29 | 2018-05-08 | 东南大学 | One kind is used for tumour cell nucleic acid aptamers screening circular card box device |
-
2016
- 2016-10-13 CN CN201621122587.4U patent/CN206114666U/en not_active Withdrawn - After Issue
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106353492A (en) * | 2016-10-13 | 2017-01-25 | 东南大学 | Cassette device for screening tumor cell aptamers |
| CN108004324A (en) * | 2017-12-29 | 2018-05-08 | 东南大学 | One kind is used for tumour cell nucleic acid aptamers screening circular card box device |
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Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| AV01 | Patent right actively abandoned |
Granted publication date: 20170419 Effective date of abandoning: 20190305 |
|
| AV01 | Patent right actively abandoned |