CN109374875A - Low noise constant temperature gravity perfusion patch clamp - Google Patents
Low noise constant temperature gravity perfusion patch clamp Download PDFInfo
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- CN109374875A CN109374875A CN201811388272.8A CN201811388272A CN109374875A CN 109374875 A CN109374875 A CN 109374875A CN 201811388272 A CN201811388272 A CN 201811388272A CN 109374875 A CN109374875 A CN 109374875A
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- Prior art keywords
- perfusate
- heating
- constant temperature
- heating liquid
- perfusion
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- 230000010412 perfusion Effects 0.000 title claims abstract description 117
- 230000005484 gravity Effects 0.000 title claims abstract description 89
- 239000007788 liquid Substances 0.000 claims abstract description 210
- 238000010438 heat treatment Methods 0.000 claims abstract description 205
- 239000012530 fluid Substances 0.000 claims description 24
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 24
- 238000003860 storage Methods 0.000 claims description 22
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 20
- 230000037452 priming Effects 0.000 claims description 13
- 238000012402 patch clamp technique Methods 0.000 claims description 7
- 239000000463 material Substances 0.000 claims description 6
- 239000008367 deionised water Substances 0.000 claims description 4
- 229910021641 deionized water Inorganic materials 0.000 claims description 4
- 238000004064 recycling Methods 0.000 claims description 3
- 125000000185 sucrose group Chemical group 0.000 claims description 3
- 238000000034 method Methods 0.000 abstract description 10
- 230000002572 peristaltic effect Effects 0.000 abstract description 6
- 238000010276 construction Methods 0.000 abstract description 2
- 230000002708 enhancing effect Effects 0.000 abstract description 2
- 238000002474 experimental method Methods 0.000 description 14
- 235000019441 ethanol Nutrition 0.000 description 8
- NJPPVKZQTLUDBO-UHFFFAOYSA-N novaluron Chemical compound C1=C(Cl)C(OC(F)(F)C(OC(F)(F)F)F)=CC=C1NC(=O)NC(=O)C1=C(F)C=CC=C1F NJPPVKZQTLUDBO-UHFFFAOYSA-N 0.000 description 7
- 229930006000 Sucrose Natural products 0.000 description 6
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 6
- 239000005720 sucrose Substances 0.000 description 6
- 230000000694 effects Effects 0.000 description 5
- 238000010586 diagram Methods 0.000 description 4
- 230000007831 electrophysiology Effects 0.000 description 4
- 238000002001 electrophysiology Methods 0.000 description 4
- 239000000523 sample Substances 0.000 description 4
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 239000011521 glass Substances 0.000 description 2
- 238000009413 insulation Methods 0.000 description 2
- 230000000149 penetrating effect Effects 0.000 description 2
- 230000005855 radiation Effects 0.000 description 2
- 239000000741 silica gel Substances 0.000 description 2
- 229910002027 silica gel Inorganic materials 0.000 description 2
- 239000007921 spray Substances 0.000 description 2
- 238000010146 3D printing Methods 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- 239000012472 biological sample Substances 0.000 description 1
- 238000001574 biopsy Methods 0.000 description 1
- 238000005520 cutting process Methods 0.000 description 1
- 125000004122 cyclic group Chemical group 0.000 description 1
- 239000012153 distilled water Substances 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 235000013399 edible fruits Nutrition 0.000 description 1
- 239000007789 gas Substances 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 238000009940 knitting Methods 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 230000010358 mechanical oscillation Effects 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 230000001376 precipitating effect Effects 0.000 description 1
- 230000002035 prolonged effect Effects 0.000 description 1
- 229920006395 saturated elastomer Polymers 0.000 description 1
- 150000003445 sucroses Chemical class 0.000 description 1
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/483—Physical analysis of biological material
- G01N33/487—Physical analysis of biological material of liquid biological material
- G01N33/48707—Physical analysis of biological material of liquid biological material by electrical means
- G01N33/48728—Investigating individual cells, e.g. by patch clamp, voltage clamp
Abstract
The present invention discloses a kind of low noise constant temperature gravity perfusion patch clamp, the perfusate that the present invention is introduced and is discharged in track using gravity and liquid tension drainage way, heat the heating liquid of track and perfusate, avoid peristaltic pump reasons for its use noise in traditional perfusion device, and the in and out records tank liquor scale of construction not etc. caused by liquid level it is unstable, so that experimental implementation process is simple, experimental result is more true and reliable；In addition, the present invention is respectively heated perfusate and perfusion slot, more stable temperature control can be obtained, perfusate is avoided to overheat, meanwhile, avoid the ambient noise enhancing and inconvenient problem of conditional electronic heating temperature control means.
Description
Technical field
The present invention relates to Medical Instruments and scientific research apparatus, suitable for the experiment of patch-clamp electro physiology the circulation of perfusate and
Administration, the constant temperature gravity perfusion patch-clamp particularly suitable for whole-cell recording technique experiment.
Background technique
In modern biology and medical domain, field, living body group especially are researched and developed in Neuroscience Research and targeted drug
The patch-clamp electrophysiological recording for knitting slice is one of extremely important and extensive experimental technique.For biopsy, usually
Oxygen environment is needed to keep cell activity, common means are to be pumped into fresh Incubating Solution using peristaltic pump to track, and pump
Extra incubation liquid out.And the unlatching of peristaltic pump would generally introduce electromagnetic noise, for high quality Patch-clamp techniques extremely
It is unfavorable；In addition, when carrying out liquid circulation by peristaltic pump, often since the flow difference of disengaging liquid causes in slot liquid level not
Stablize even liquid to overflow.Therefore, a kind of energy is needed to provide the solution of low background noise and steady liquid level in Patch-clamp techniques experiment
Certainly scheme.Meanwhile in an experiment, it usually needs maintain the constant temperature of experiment sample, be generally higher than the physiological temp of room temperature, at present
Common technology is to heat before perfusate flows into track to it, and the method usually requires heating perfusate to being much higher than work
Temperature is lost with heat compensation, and the fluid temperature in perfusion tube and track is avoided to be lower than the operating temperature of minimum permission.This
Outside, perfusate generally comprises precipitable ion, can be precipitated due to overheat, and then influences experiment；Another heat compensation damage
The technology of mistake is to heat track by electrothermal method, and such method can be introduced directly into electromagnetic noise, be unfavorable for electrophysiological recording.
Therefore, it needs a kind of to provide the solution party of low background noise, isoperibol and steady liquid level simultaneously in Patch-clamp techniques experiment
Case.
Summary of the invention
A kind of low noise constant temperature gravity perfusion Patch Clamp System is provided with the present invention to realize the purpose for solving disadvantages mentioned above.
In order to achieve the above objectives, solution of the invention is to provide a kind of low noise constant temperature gravity perfusion patch-clamp dress
It sets, comprising:
Constant temperature gravity perfusion slot, the constant temperature gravity perfusion slot are arranged on patch-clamp micromanipulation platform；
Add hydrothermal circulation bag, for storing heating liquid；
Liquid collection bag is heated, for recycling heating liquid；
Liquid circulation pipe is heated, the heating liquid circulation pipe is connected to the constant temperature gravity perfusion slot, described plus hydrothermal circulation
Between bag and the heating liquid collection bag, so that heating liquid is in the constant temperature gravity perfusion slot, described plus hydrothermal circulation bag and institute
State heating liquid collection bag internal circulation flow；
Perfusate drop bottle, for storing perfusate；
Perfusion fluid catheter, the perfusion fluid catheter are connected between perfusate drop bottle and the constant temperature gravity perfusion slot, are filled
Flow liquid injects constant temperature gravity perfusion slot by gravity.
According to one embodiment of present invention, wherein the constant temperature gravity perfusion slot includes:
Track, for carrying out Patch-clamp techniques；
Drain duct, for perfusate to be discharged；
Drainage trough is arranged between track and drain duct, for guiding perfusate from track to drain duct
Flowing；With
Liquid circulating line is heated, the heating liquid circulating line surrounds the track, the drainage trough and the drain
Duct setting, heating liquid circulate in the heating liquid circulating line.
According to one embodiment of present invention, wherein further include:
The two sides of the constant temperature gravity perfusion slot, the constant temperature is arranged in perfusion slot fixed-wing, the perfusion slot fixed-wing
Gravity perfusion slot is fixed by the perfusion slot fixed-wing and the patch-clamp micromanipulation platform.
According to one embodiment of present invention, wherein further include:
Perfusate heating muff, the perfusate heating muff are arranged around the perfusion fluid catheter, for heating in institute
State the perfusate flowed in perfusion fluid catheter；With
Heating storage tanks, the heating storage tanks are arranged on the perfusate heating muff, for storing and heating
Heating liquid in the perfusate heating muff.
According to one embodiment of present invention, wherein the track is circular strip inclined-plane reverse frustoconic and includes:
Perfusate inlet opening, the perfusate inlet opening to connection perfusion fluid catheter；
The side of perfusate water conservancy diversion rib, the perfusate water conservancy diversion rib is arranged close to perfusate inlet opening, and the perfusate is led
The other side of stream rib is provided with half the height opening, and extra perfusate can be overflowed by described half high opening.
According to one embodiment of present invention, wherein the level height in the track side of the drainage trough is higher than
The level height in drain duct side of the drainage trough, so that the drainage trough has from the track towards institute
State the inclined inclined surface in drain duct.
According to one embodiment of present invention, described wherein being provided with protrusion drainage rib on the surface of the drainage trough
Drainage rib extends into the drain duct along the surface of the drainage trough, and the surface of the drainage trough be frosted not
Smooth surface.
According to one embodiment of present invention, wherein the heating liquid circulating line in turn includes:
Heat liquid entrance；
Half of section of track is surrounded clockwise；
The section in drain duct is extended to from track；
Section around drain duct；
The section of track is extended to from drain duct；
Half of section of track is surrounded counterclockwise；With
Heat liquid outlet；
Cutting for the pipeline of half of section of track is wherein surrounded around half of section of track and counterclockwise clockwise
Face is triangle, and circulation has the heating liquid for being not likely to produce bubble in pipeline.
According to one embodiment of present invention, wherein the heating liquid is deionized water, saturation sucrose solution or high-purity
Ethyl alcohol.
According to one embodiment of present invention, wherein heating liquid circulation bag includes:
The heating liquid storage bag being fabricated from a flexible material；
Hard base；And
Top cover is provided with exhaust valve on the top cover；
It wherein is provided with temperature control heating pipe on the direction of the direction heating liquid storage bag of the hard base, described hard
It is provided with water level control type self priming pump in matter pedestal, while the lower part of the hard being provided with heating liquid delivery outlet, heats liquid
It is conveyed by self gravity from heating liquid delivery outlet.
Compared with prior art, the present invention at least has the following technical effect that
1. perfusion fully relies on its own gravity in track and surface tension of liquid flows into and discharge, it can achieve note
It is consistent to record disengaging fluid flow in slot, and then realizes that liquid level is stablized in track.
2. perfusion fully relies on its own gravity in track and surface tension of liquid flows into and discharge, peristaltic pump is got rid of
Etc. the auxiliary of all kinds of machinery, the noise jamming of additional electronic component introducing is avoided.
3. heating using heating liquid to perfusate and track, the noise jamming of additional electronic component introducing is got rid of.
4. perfusate and heating tank use independent heating system respectively, temperature can be carried out respectively to perfusate and heating tank
Degree control can be avoided excessively heating perfusate while meeting operating temperature, obtain more stable temperature control.
5. track heats liquid using completely enclosed circulation, reduce noise, mechanical oscillation and the generation for heating bubble.
6. heat liquid storage bag in have water level control type self priming pump, can automatically keep it is enough, have gravitional force
Heating liquid.Meanwhile self priming pump can carry out manual switch control, to reduce the electromagnetic noise in high electrical accuracies experiment.
7. heating liquid uses pure water, it is saturated sucrose solution or ethyl alcohol, can further avoid being detained in track heating tube
Bubble.
8. perfusion slot is made of 3D printing method, manufacturing process is convenient, and cost is relatively low.
Detailed description of the invention
Fig. 1 is the stereoscopic schematic diagram according to the structure of the whole system of the embodiment of the present invention.
Fig. 2 is the schematic top plan view according to the structure of the whole system of the embodiment of the present invention.
Fig. 3 is the schematic elevation view according to the structure of the constant temperature gravity perfusion slot of the embodiment of the present invention.
Fig. 4 is the stereoscopic schematic diagram according to the structure of the constant temperature gravity perfusion slot of the embodiment of the present invention.
Fig. 5 is the schematic top plan view according to the structure of the constant temperature gravity perfusion slot of the embodiment of the present invention.
Fig. 6 is the stereoscopic schematic diagram according to the embodiment of the present invention plus hydrothermal circulation bag the structure.
Fig. 7 is the schematic top plan view according to the embodiment of the present invention plus hydrothermal circulation bag the structure.
Fig. 8 is the stereoscopic schematic diagram according to the structure of the heating liquid collection bag of the embodiment of the present invention.
Fig. 9 is the schematic top plan view according to the structure of the heating liquid collection bag of the embodiment of the present invention.
Appended drawing reference:
1, constant temperature gravity perfusion slot；
11, track；
111, perfusate inlet opening；
112, perfusate drain rib；
113, drain is open；
12, drainage trough；
121, drainage trough drains rib；
122, drainage trough matte surface；
13, drain duct；
131, drain duct drains rib；
132, Drainage pipe interface；
14, liquid circulating line is heated；
141, liquid entrance is heated；
142, heating liquid outlet；
15, perfusion slot fixed-wing；
151, perfusion slot fixation hole；
16, attachment fixation hole；
2, add hydrothermal circulation bag；
201, liquid storage bag is heated；
202, pedestal；
203, top cover；
204, exhaust valve；
205, temperature control heating pipe；
206, water level control self priming pump；
207, add the heating liquid delivery outlet of hydrothermal circulation bag；
208, add the heating liquid suction inlet of hydrothermal circulation bag；
209 heating liquid pump entrances；
210, add the heating liquid outlet of hydrothermal circulation bag；
3, liquid collection bag is heated；
31, liquid storage bag is heated；
32, pedestal；
33, top cover；
34, exhaust valve；
35, the heating liquid input port of liquid collection bag is heated；
36, the heating liquid delivery outlet of liquid collection bag is heated；
37, the heating liquid outlet of liquid collection bag is heated；
4, liquid circulation pipe is heated；
41, the first heating liquid circulation pipe；
42, the second heating liquid circulation pipe；
43, third heats liquid circulation pipe；
5, perfusate heating muff；
6, heated at constant temperature case；
7, perfusate drop bottle；
8, perfusion fluid catheter；
9, perfusate discharge pipe.
Specific embodiment
For technical solution of the present invention is explained further, the present invention is explained in detail below by specific embodiment
It states.
The present invention discloses a kind of low noise constant temperature gravity perfusion Patch Clamp System comprising one is arranged in patch-clamp micromanipulation
Constant temperature gravity perfusion slot 1 on platform, constant temperature gravity perfusion slot of the present invention have the track for carrying out Patch-clamp techniques, discharge
The drainage trough of perfusate and drain duct for perfusate to be discharged；It further include one plus hydrothermal circulation bag 2, a heating liquid in system
Collection bag 3, and connection constant temperature gravity perfusion slot plus hydrothermal circulation bag, the heating liquid circulation pipe 4 for heating liquid collection bag three,
One is arranged in the perfusate heating muff 5 on perfusion tube, the heating storage tanks 6 of storage and heating perfusate heating liquid, and system is led to
Crossing gravity makes perfusate inject constant temperature gravity perfusion slot 1 by perfusate drop bottle 7 and perfusion fluid catheter 8, and passes through gravity
And liquid tension effect, by 9 discharge system of perfusate discharge pipe；The use of circulating-heating liquid can be avoided bubble generation in system
Liquid.
The perfusate that the present invention is introduced and is discharged in track using gravity and liquid tension drainage way, heating record
The heating liquid of slot and perfusate avoids peristaltic pump reasons for its use noise and in and out records tank liquor in traditional perfusion device
Liquid level caused by the scale of construction is not equal is unstable, so that experimental implementation process is simple, experimental result is more true and reliable；In addition, of the invention
Perfusate and perfusion slot are respectively heated, more stable temperature control can be obtained, perfusate is avoided to overheat, meanwhile, it avoids
The ambient noise enhancing of conditional electronic heating temperature control means and inconvenient problem.
According to one embodiment of present invention, a kind of low noise constant temperature gravity perfusion Patch Clamp System a comprising setting exists
Constant temperature gravity perfusion slot 1 on patch-clamp micromanipulation platform, constant temperature gravity perfusion slot of the present invention, which has, carries out patch-clamp note
The track 11 of record is discharged the drainage trough 12 of perfusate and the drain duct 13 for perfusate to be discharged, and adds for recycling
The heating liquid circulating line 14 of hydrothermal solution, in addition, perfusion slot fixed-wing 15 and micro- behaviour of the constant temperature gravity perfusion slot by two sides
It is fixed as platform, and has attachment fixation hole 16；It further include one plus hydrothermal circulation bag 2 in system, one heats liquid collection bag 3, and
Constant temperature gravity perfusion slot plus hydrothermal circulation bag, the heating liquid circulation pipe 4 for heating liquid collection bag three are connected, heats liquid in system
Internal circulation flow；One is arranged in the perfusate heating muff 5 on perfusion tube, the heating storage of storage and heating perfusate heating liquid
Case 6, system make perfusate inject constant temperature gravity perfusion slot 1 by perfusate drop bottle 7 and perfusion fluid catheter 8 by gravity,
And acted on by gravity and liquid tension, by 9 discharge system of perfusate discharge pipe.
According to one embodiment of present invention, wherein track 11 is circular strip inclined-plane reverse frustoconic, and there is a connection to fill
The perfusate inlet opening 111 of flow liquid conduit 8, close to perfusate inlet opening at be equipped with a perfusate water conservancy diversion rib 112, and positioned at leading
Half high opening 113 at the one of rib opposite side is flowed, record trench bottom pastes circular glass coverslip, for holding perfusate, excessive
Perfusate is overflowed by the high opening of track half.
12 linkage record channel opening 111 of drainage trough and drain duct 13, drainage trough slot have inclined ramp, drainage trough record
Channel opening direction is higher than drain duct direction, and drainage rooved face has protrusion drainage rib 121, and drainage rib prolongs along drainage trough lower surface
Drainage hole 3 is extended into, drainage rooved face is the rough surface 122 of frosted.
13 center of drain duct and 11 centre distance of track are greater than 40mm, and drain duct is penetrating duct vertically downward,
Drainage hole is that diameter is greater than 6mm circle duct, and drainage hole road has protrusion drainage rib, and drainage rib 131 drains rib 121 by drainage trough
Extend, it is parallel with drain internal surface of hole, drain internal surface of hole is raised in close to track end, and 13 lower end of drain duct extends to row
Liquid pipe pipeline joint 132, for connecting perfusate discharge pipe 9.
Heating liquid circulating line 14 starts from a heating liquid entrance 141, surrounds track 11, and prolonged by record channel opening 111
13 direction of drain duct is extended to, it is oppositely extending to record channel opening 111 again around drain duct 13, track 11 is surrounded, is connected to
Another heating liquid outlet 142；It is wherein triangular-section pipeline around 11 part of track, circulation is not likely to produce gas in pipeline
The heating liquid of bubble.
Adding hydrothermal circulation bag 2 includes heating liquid storage bag 201, hard base 202 and the top cover being fabricated from a flexible material
203 main bodys constituted, have exhaust valve 204 on top cover, and pedestal is provided with temperature control heating pipe towards heating liquid storage bag direction
205, water level control type self priming pump 206 is provided in pedestal, while lower part being provided with heating liquid delivery outlet 207, liquid passes through
Self gravity from heating liquid delivery outlet 207 to system other parts convey, water level control type self priming pump 206 be set on the base
Connected by heating liquid suction inlet 208, the heating liquid for being circulated in system is pumped into the pump housing by self priming pump, pass through heating liquid pump entrance
209, which leave the pump housing, enters heating liquid storage bag, in addition, bottom is provided with heating liquid outlet 210 on heated base, for arranging
Dry heating liquid.
Heating liquid collection bag 3 includes heating liquid storage bag 31, hard base 32 and the top cover 33 being fabricated from a flexible material
The main body of composition has exhaust valve 34 on top cover, heats liquid input port 35, heats liquid delivery outlet 36, pedestal bottom is provided with
Liquid outlet 37 is heated, for draining heating liquid.
The silica gel hose that liquid circulation pipe 4 and perfusate heating muff 5 select wall thickness to be not less than 1mm is heated, and outside pipeline
Wall surrounds or sprays heat radiation reflective insulation layer.
Heated at constant temperature case 6 is reequiped by water-bath, is had the outlet of heating hydrorrhea, be may connect on heating liquid circulation pipe 4.
Perfusion fluid catheter 8 is the flexible hose that tube wall is less than 0.2mm, and outer diameter is less than 5 internal diameter 2mm of heating muff or more.
Heating muff 5 surrounds perfusion fluid catheter 8, and in heating liquid of the two pipeline gap perfusions from heated at constant temperature case 6, with
Heat the perfusate in perfusion fluid catheter 8.
Perfusate discharge pipe 9 is connected to Drainage pipe interface 132, is not more than 132 outer diameter of Drainage pipe interface, wall for internal diameter
Thickness is not less than the flexible hose of 0.5mm.
Heating liquid is preferably deionized water, is saturated sucrose solution or high purity ethanol.
Referring to Fig.1 shown in -5, the perfusate for electro physiology experiment in perfusate drop bottle 7 is stored in by self gravity
Constant temperature gravity perfusion slot 1 is injected via perfusion fluid catheter 8.Constant temperature gravity perfusion slot 1 is for holding electro physiology sample.Particularly,
Electro physiology sample is provided in the track 11 of constant temperature gravity perfusion slot 1.In constant temperature gravity perfusion slot 1, perfusate passes through
The perfusate inlet opening 111 of constant temperature gravity perfusion slot 1 flows into the track 11 of constant temperature gravity perfusion slot 1, with to being placed on note
Biological sample in record slot 11 provides fresh perfusate.Excessive perfusate is overflowed via half high opening 113 of track 11,
And enters drain duct 13 by drainage trough 12, be then discharged by perfusate discharge pipe 9.
According to one embodiment of present invention, low noise constant temperature gravity perfusion patch-clamp includes being arranged in patch-clamp micromanipulation
Constant temperature gravity perfusion slot 1 on platform.According to one embodiment of present invention, constant temperature gravity perfusion slot 1 passes through the perfusion slot of two sides
Fixed-wing 15 is fixed with micromanipulation platform.And as shown in figure 5, being additionally provided with multiple fixation holes on constant temperature gravity perfusion slot 1
16, for constant temperature gravity perfusion slot 1 to be fixed to micromanipulation platform.
Constant temperature gravity perfusion slot 1 has the track 11 for carrying out Patch-clamp techniques, for guiding perfusate that track is discharged
11 drainage trough 12, the drain duct 13 for perfusate to be discharged and the heating liquid circulating line 14 for circulating-heating liquid.
As shown in Fig. 3, track 11 is circular strip inclined-plane reverse frustoconic.According to one embodiment of present invention, perfusion
Fluid catheter 8 is connected to track 11 by perfusate inlet opening 111.In track 11 close to setting at perfusate inlet opening 111
It is equipped with perfusate water conservancy diversion rib 112, perfusate is led into track 11.In track 11 and perfusate inlet opening 111
Half high opening 113 is provided at opposite side.Circular glass coverslip is pasted at the bottom of track 11, for holding
Perfusate.The half 113 overflow record slots 11 of high opening that excessive perfusate passes through track 11.The perfusate of overflow record slot 11
Discharge track 11 is guided by drainage trough 12, and enters perfusate discharge pipe 9 via drain duct 13.
As shown in figure 5,12 linkage record channel opening 111 of drainage trough and drain duct 13,12 slot of drainage trough has inclination oblique
Face, horizontal position of the drainage trough 12 at neighbouring track 11 are higher than horizontal position of the drainage trough 12 at neighbouring drain duct 13
It sets, the perfusate of overflow record slot 11 is enabled to flow to drain duct 13 from track 11.In addition, on the surface of drainage trough 12
It is provided with protrusion drainage rib 121, rib 121 is drained along the surface of drainage trough 12 and extends into drain duct 13, drainage trough 12
Surface is the rough surface 122 of frosted.
According to one embodiment of present invention, the distance between the center in drain duct 13 and the center of track 11 are greater than
40mm, drain duct 13 are that penetrating duct, drain duct 13 are the round duct that diameter is greater than 6mm vertically downward.Drain duct
Can have protrusion drainage rib 131, drainage rib 131 from drainage trough protrusion drainage rib 121 extend, and with drain duct 13
Inner surface is parallel, is raised in the close track end of the inner surface in drain duct.The lower end in drain duct 13 extends to drain pipe
Pipeline joint 132, for being connected to perfusate discharge pipe 9.
As shown in figure 5, being surrounded in the periphery of the track 11 of constant temperature gravity perfusion slot 1 by heating liquid circulating line 14.Add
Hydrothermal circulation pipeline 14 starts from heating liquid entrance 141, the direction as guided by arrow in Fig. 5, around the half of track 11
It advances, is extended through record channel opening 111 to 13 direction of drain duct, surround drain duct 13, it is oppositely extending to return to track
Opening 111 eventually terminates at heating liquid outlet 142 around the other half of track 11.It is followed in the heating liquid circulating line 14
Ring has the heating liquid for being not likely to produce bubble.
According to one embodiment of present invention, the part around track 11 for heating liquid circulating line 14 has triangle
Section.
Referring to shown in Fig. 2, constant temperature gravity perfusion slot 1 plus hydrothermal circulation bag 2 and heating 3 three of liquid collection bag are by adding
Hydrothermal circulation pipe 4 and be fluidly connected to one another each other.According to one embodiment of present invention, heating liquid circulation pipe 4 includes
Three independent connecting tubes: the first heating liquid circulation pipe 41, second heats liquid circulation pipe 42 and third heats liquid circulation pipe 43.The
One heating liquid circulation pipe 41 setting is in constant temperature gravity perfusion slot 1 and adds between hydrothermal circulation bag 2；Second heating liquid circulation pipe 42 is set
It sets between constant temperature gravity perfusion slot 1 and heating liquid collection bag 3；The third heating setting of liquid circulation pipe 43 is adding hydrothermal circulation bag 2
Between heating liquid collection bag 3.The heating liquid entrance 141 of circulating line 14 is connected to heating by the first heating liquid circulation pipe 41
The heating liquid delivery outlet 207 of liquid circulation bag 2.The heating liquid outlet 142 of circulating line 14 is connected by the second heating liquid circulation pipe 42
It is connected to the heating liquid input port 35 of heating liquid collection bag 3.The heating liquid suction inlet 28 of hydrothermal circulation bag 2 is added to heat liquid by third
Circulation pipe 43 is connected to the heating liquid delivery outlet 36 of heating liquid collection bag 3.Heat flow direction of the liquid in heating liquid circulation pipe 4
As shown in the arrow in attached drawing 2, firstly, heating liquid from add hydrothermal circulation bag 2 by first heating liquid circulation pipe 41 flow to perseverance
Then warm gravity perfusion slot 1 heats liquid from constant temperature gravity perfusion slot 1 and flow to heating liquid time by the second heating liquid circulation pipe 42
Closing bag 3, heating liquid, which is flow to from heating liquid collection bag 3 by third heating liquid circulation pipe 43, again adds hydrothermal circulation bag 2.
According to one embodiment of present invention, heating liquid selected from pure water, saturation sucrose solution or ethyl alcohol it is a kind of or
Its any combination.
Shown in referring to Fig.1, perfusate is in the heating by being flowed in perfusate heating muff 5 when perfusion fluid catheter 8
Liquid heats, and the heating liquid in perfusate heating muff 5 is flowed out by its own gravity from heated at constant temperature case 6.
According to one embodiment of present invention, general distilled water can be used in the heating liquid in perfusate heating muff 5.
As shown in fig. 6-7, adding hydrothermal circulation bag 2 includes: the heating liquid storage bag 201 being fabricated from a flexible material, hard bottom
Seat 202 and top cover 203 constitute main body；Exhaust valve 204 on top cover is set；It is stored in hard base 202 towards heating liquid
The temperature control heating pipe 205 that 201 sides of bag set up；The water level control type self priming pump 206 being arranged in hard base 202；Hard
The heating liquid delivery outlet 207 of 202 lower part of matter pedestal setting.Liquid by self gravity from heating liquid delivery outlet 207 to system its
Its part conveys, and water level control type self priming pump 206 is connect with the heating liquid suction inlet 208 being set on the base, and is circulated in system
Heating liquid the pump housing is pumped by self priming pump 206, and leave the pump housing by heating liquid pump entrance 209 and enter heating liquid storage bag 2.
In addition, heating liquid outlet 210 is provided at the bottom of hard base 202, for draining heating liquid.
As Figure 8-9, heating liquid collection bag 3 includes: heating liquid storage bag 31, the hard base being fabricated from a flexible material
32 and top cover 33 constitute main body；Exhaust valve 34, heating liquid input port 35 and heating liquid delivery outlet 36 on top cover are set；
Heating liquid outlet 37 is provided at 32 bottom of hard base, for draining heating liquid.
According to one embodiment of present invention, liquid circulation pipe 4 is heated, perfusate heating muff 5 selects wall thickness to be not less than 1mm
Silica gel hose, and surround or spray heat radiation reflective insulation layer in pipeline outer wall.
According to one embodiment of present invention, heated at constant temperature case 6 is reequiped by water-bath, is had the outlet of heating hydrorrhea, can be connected
It is connected on heating liquid circulation pipe 4.
According to one embodiment of present invention, perfusion fluid catheter 8 is tube wall less than (flexible hose of 0.2mm, outer diameter are small
In the internal diameter 2mm or more of heating muff 5.
According to one embodiment of present invention, heating muff 5 surrounds perfusion fluid catheter 8, and in two pipeline gap perfusions
Heating liquid from heated at constant temperature case 6, to heat the perfusate in perfusion fluid catheter 8.
According to one embodiment of present invention, perfusate discharge pipe 9 is connected to Drainage pipe interface 132, perfusate discharge
Pipe 9 is flexible hose of its internal diameter no more than the outer diameter of Drainage pipe interface 132.
According to one embodiment of present invention, perfusate discharge pipe 9 is the flexible hose that wall thickness is not less than 0.5mm.
According to one embodiment of present invention, heating liquid is preferably deionized water, saturation sucrose solution or high purity ethanol.
According to one embodiment of present invention, temperature control process is as follows:
Perfusate is preheated to slightly above before entering constant temperature gravity perfusion slot 1 or is equal to operating temperature.Perfusate
After entering gravity perfusion slot 1, continue to heat and heating the heating liquid flowed in liquid circulating line 14, so that filling
The temperature of flow liquid maintains at operating temperature.
Specifically, the heating temperature in heated at constant temperature case 6 is adjusted to be equal to or slightly higher than perfusate operating temperature.This is
Since excessively high temperature can make perfusate generate precipitating and be not easy to control, even results in sample and inactivate at high temperature, then, root
It, will be warm according to the heating temperature of the temperature control heating pipe 205 in the adjustment of the actual conditions such as the flow velocity of perfusion and room temperature plus hydrothermal circulation bag 2
Degree is set as not less than operating temperature.
According to one embodiment of present invention, the heating liquid cyclic process in constant temperature gravity perfusion slot 1 is as follows:
It is stored in and adds the heating liquid in hydrothermal circulation bag 2 by its gravity, first inflow constant temperature gravity perfusion slot 1, then again
Heating liquid collection bag 3 is flowed into from constant temperature gravity perfusion slot 1.According to one embodiment of present invention, it sets adding in hydrothermal circulation bag 2
It is equipped with self priming pump.Self priming pump manually or electrically starts, will heat the heating liquid in liquid collection bag 3 during experimental record gap
In pump sucking plus hydrothermal circulation bag 2, to complete to form the circulation of heating liquid.
It is according to an embodiment of the invention that the use that not only can be reduced electronic equipment in experimental record is set, but also can make to add
Hydrothermal solution carries out closed circulation, and bubble is avoided to generate, and simplifies the operation in experiment.
The heating liquid used according to one embodiment of present invention is pure water or saturation sucrose solution or ethyl alcohol, is had
Preferable Anti-bubble function.Particularly, the Anti-bubble effect of ethyl alcohol is better than saturation sucrose solution, is saturated the Anti-bubble effect of sucrose solution
Fruit is better than pure water.But ethyl alcohol is more demanding to seal for pipe joints, and it is saturated sucrose solution microorganism easy to breed, therefore,
It can need to be selected according to requirement of experiment in experiment.
According to one embodiment of present invention, those skilled in the art can according to need packet known to voluntarily selection
Include structure and technological means commonly used by other patch clamp experiments including Incubating Solution circulation path.
Claims (10)
1. a kind of low noise constant temperature gravity perfusion patch clamp, comprising:
Constant temperature gravity perfusion slot (1), the constant temperature gravity perfusion slot (1) are arranged on patch-clamp micromanipulation platform；
Add hydrothermal circulation bag (2), for storing heating liquid；
It heats liquid collection bag (3), for recycling heating liquid；
It heats liquid circulation pipe (4), the heating liquid circulation pipe (4) is connected to the constant temperature gravity perfusion slot (1), the heating liquid
It recycles between bag (2) and the heating liquid collection bag (3), so that heating liquid is in the constant temperature gravity perfusion slot (1), the heating
Liquid recycles bag (2) and heating liquid collection bag (3) internal circulation flow；
Perfusate drop bottle (7), for storing perfusate；
Perfusion fluid catheter (8), the perfusion fluid catheter (8) are connected to perfusate drop bottle (7) and the constant temperature gravity perfusion slot (1)
Between, perfusate injects constant temperature gravity perfusion slot (1) by gravity.
2. low noise constant temperature gravity perfusion patch clamp according to claim 1, wherein the constant temperature gravity perfusion slot (1)
Include:
Track (11), for carrying out Patch-clamp techniques；
Drain duct (13), for perfusate to be discharged；
Drainage trough (12) is arranged between track (11) and drain duct (13), for guiding perfusate from track (11)
It is flowed to drain duct (13)；
With heating liquid circulating line (14), the heating liquid circulating line (14) surrounds the track (11), the drainage trough
(12) it is circulated in the heating liquid circulating line (14) with the drain duct (13) setting, heating liquid.
3. low noise constant temperature gravity perfusion patch clamp according to claim 1, wherein further include:
Perfusion slot fixed-wing (15), the perfusion slot fixed-wing (15) are arranged in the two sides of the constant temperature gravity perfusion slot (1), institute
It states constant temperature gravity perfusion slot (1) and is fixed by the perfusion slot fixed-wing (15) with the patch-clamp micromanipulation platform.
4. low noise constant temperature gravity perfusion patch clamp according to claim 1, wherein further include:
Perfusate heating muff (5), the perfusate heating muff (5) is arranged around the perfusion fluid catheter (8), for heating
The perfusate flowed in the perfusion fluid catheter (8)；
With heating storage tanks (6), the heating storage tanks (6) are arranged on the perfusate heating muff (5), for storing
With heating liquid of the heating in the perfusate heating muff (5).
5. low noise constant temperature gravity perfusion patch clamp according to claim 2, wherein the track (11) is circle
With inclined-plane reverse frustoconic and include:
Perfusate inlet opening (111), the perfusate inlet opening (111) to connection perfusion fluid catheter (8)；
The side of perfusate water conservancy diversion rib (112), the perfusate water conservancy diversion rib (112) is arranged close to perfusate inlet opening (111), institute
The other side for stating perfusate water conservancy diversion rib (112) is provided with half the height opening (113), and extra perfusate can be high by described half
Opening is overflowed.
6. low noise constant temperature gravity perfusion patch clamp according to claim 2, wherein the drainage trough (12) in institute
The level height for stating track (11) side is higher than the level height in drain duct (13) side of the drainage trough (12)
Degree, so that the drainage trough (12) has from the track (11) towards the inclined inclined surface of the drain duct (13).
7. low noise constant temperature gravity perfusion patch clamp according to claim 6, wherein the table in the drainage trough (12)
Protrusion drainage rib (121) is provided on face, drainage rib (121) extends into described along the surface of the drainage trough (12)
Drain duct (13), and the surface of the drainage trough (12) is the rough surface (122) of frosted.
8. low noise constant temperature gravity perfusion patch clamp according to claim 6, wherein the heating liquid circulating line
(14) in turn include:
It heats liquid entrance (141)；
Half of section of track (11) is surrounded clockwise；
The section of drain duct (13) is extended to from track (11)；
Section around drain duct (13)；
The section of track (11) is extended to from drain duct (13)；
Half of section of track (11) is surrounded counterclockwise；With
Heat liquid outlet (142)；
Wherein clockwise around half of section of track (11) and the pipeline of half of section counterclockwise around track (11)
Section be triangle, and in pipeline circulation have the heating liquid for being not likely to produce bubble.
9. low noise constant temperature gravity perfusion patch clamp according to claim 1, wherein the heating liquid be deionized water,
It is saturated sucrose solution or high purity ethanol.
10. low noise constant temperature gravity perfusion patch clamp according to claim 1, wherein heating liquid circulation bag (2)
Include:
The heating liquid storage bag (201) being fabricated from a flexible material；
Hard base (202)；And
Top cover (203) is not equipped with exhaust valve (204) on the top cover；
Temperature control heating pipe (205) wherein are provided on the direction of direction heating liquid storage bag (201) of the hard base,
Be provided in the hard base water level control type self priming pump (206), at the same the lower part of the hard be provided with heating liquid it is defeated
It exports (207), heating liquid is conveyed by self gravity from heating liquid delivery outlet (207).
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811388272.8A CN109374875A (en) | 2018-11-20 | 2018-11-20 | Low noise constant temperature gravity perfusion patch clamp |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811388272.8A CN109374875A (en) | 2018-11-20 | 2018-11-20 | Low noise constant temperature gravity perfusion patch clamp |
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| CN109374875A true CN109374875A (en) | 2019-02-22 |
Family
ID=65376789
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|---|---|---|---|
| CN201811388272.8A Pending CN109374875A (en) | 2018-11-20 | 2018-11-20 | Low noise constant temperature gravity perfusion patch clamp |
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| CN112056306A (en) * | 2020-06-22 | 2020-12-11 | 南京易太斯生物科技有限公司 | Brain slice perfusion tank suitable for electrophysiological experiments |
| CN113406316A (en) * | 2021-06-17 | 2021-09-17 | 重庆医科大学附属儿童医院 | Electrophysiological patch clamp perfusion device |
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| CN113406316A (en) * | 2021-06-17 | 2021-09-17 | 重庆医科大学附属儿童医院 | Electrophysiological patch clamp perfusion device |
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