CN102151997A - Method for processing micropore of patch clamp chip - Google Patents
Method for processing micropore of patch clamp chip Download PDFInfo
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- CN102151997A CN102151997A CN2011100347475A CN201110034747A CN102151997A CN 102151997 A CN102151997 A CN 102151997A CN 2011100347475 A CN2011100347475 A CN 2011100347475A CN 201110034747 A CN201110034747 A CN 201110034747A CN 102151997 A CN102151997 A CN 102151997A
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Abstract
The invention discloses a method for processing a micropore of a patch clamp chip. The method comprises the following steps of: 1) expanding an ultraviolet Gaussian laser beam which is emitted by an ultraviolet laser and has the power of 1 to 7W by using a beam expander; 2) taking a central light spot of the ultraviolet Gaussian laser beam with an aperture of 1.2 to 1.8 millimeters out through an adjusting diaphragm from the ultraviolet Gaussian laser beam expanded by the beam expander; and 3) focusing the central light spot of the ultraviolet Gaussian laser beam to the surface of the glass patch clamp chip for perforation, and during perforation, forming a funnel-shaped micropore on the glass patch clamp chip by controlling a central diameter of the adjusting diaphragm in the step 2). By the method, the high energy of the focused ultraviolet laser can break a molecular structure of a glass material on a patch clamp chip clamp within an extreme time, and uniform pores can be formed on the glass material.
Description
Technical field
The present invention relates to the laser micropore processing technique field, be specifically related to a kind of patch clamp chips capillary processing method.
Technical background
Patch clamp technique is the important technology in the ion channel research.The advantage that information content is big, resolution ratio is high though classical patch-clamp possesses skills, drawbacks limit such as flux is low, operation is loaded down with trivial details, requirement for experiment condition height its application in researchs such as cell physiological and drug discovery.Therefore the patch clamp chips technology is arisen at the historic moment.The patch clamp chips technology can carry out high-throughout electrophysiologicalmeasurements measurements, form the high impedance sealing-in automatically, and can combine with the optical measurement means, and it has promoted the research of ion channel and high-flux medicaments sifting.The patch clamp chips technology adopts the planar structure that has the micron order hole to replace glass microelectrode (as shown in Figure 1) and cell to form sealing-in, only need cell suspension is dropped on the hole, apply a negative pressure or an electrostatic field, cell is directed on the hole, applies a negative pressure again and can make cell and chip chamber form the high impedance sealing-in.More convenient to operate like this, quick.And can be that electrod-array carries out record simultaneously to a plurality of cells with chip manufacturing.The micropore of above-mentioned diaphragm embedding chip is leak shape (as shown in Figure 2), and the entrance hole diameter of micropore is greater than the outlet aperture, and the scope of micropore entrance hole diameter is 2～10 μ m, and the scope of micropore outlet aperture is 1～2 μ m, and this structure helps the perfusion of cell suspension.
The material of the making patch clamp chips that has adopted at present, has silicon, quartz crystal or glass and organic polymer body.There is a series of problem based on the patch clamp chips of silicon materials substrate fabrication in employing in the punching experiment, be difficult to form above-mentioned leak shape micropore; The patch clamp chips of utilizing quartz crystal good insulation performance character to make, can use the plane process technology of standard, obtain the hole of micron to submicron order, the performance parameter of finding this patch clamp chips in the experiment is fine, but the shape of the hole of getting is triangle always, makes cell and chip chamber be difficult to form the sealing-in of high impedance; Glass and organic polymer body also are good insulating materials, but also have transparent, the convenient advantage of observing and can combine with other optical detection means such as detection technique of fluorescence, glass membrane pincers chip and cell can form the high impedance sealing-in preferably.In addition, glass has good electricity and engineering properties, surperficial possess hydrophilic property, is more satisfactory diaphragm embedding chip material.
Method at processing micropore on glass adopts single ion to follow the trail of lithographic technique usually at present, and the hole diameter of processing on chip can reach 1 μ m, even littler, and hole is mellow and full, smooth.But this method efficient is lower, and it is bigger to promote difficulty.Therefore how be the key of large-scale application patch clamp chips technology at the leak shape micropore that processes 1～10 μ m aperture on glass.
As everyone knows, laser boring processing is the important component part of laser processing technology, and laser boring is compared with other drilling methods, and maximum difference is the characteristics that it has high speed, high accuracy and high-adaptability.In addition, it also has, and punching is even, the heat affected area is little, add the noiseless in man-hour, joint-cutting edge-perpendicular degree is good, it is smooth to cut edge, process realizes advantages such as automation control easily, but the aperture of present laser drilling minimum process and is difficult to get leak shape micropore about 100 μ m.In addition, traditional YAG laser and CO
2Laser is difficult to act on glass material, the weak effect of processing, and also the glass of being processed is easy to crack and collapse the limit, can not satisfy the requirement of diaphragm embedding chip manufacture.Therefore how the processing that laser drilling is applied to diaphragm embedding chip is the key issue that the present invention will solve.
Summary of the invention
The objective of the invention is at above-mentioned technical problem, a kind of patch clamp chips capillary processing method that can be implemented in processing micron order hole on glass is provided, make it can be applicable to large-scale industrialization production.
For realizing this purpose, a kind of patch clamp chips capillary processing method that the present invention is designed is characterized in that it comprises the steps:
The power that step 1) is sent ultraviolet laser is 1～7 watt ultraviolet Gaussian laser beam, expands bundle by beam expanding lens;
Step 2) the ultraviolet Gaussian laser beam that beam expanding lens is expanded after restrainting is the center spot of the ultraviolet Gaussian laser beam of 1.2～1.8mm by regulating aperture taking-up beam diameter scope;
Step 3) focuses on glass membrane pincers chip surface with the center spot of described ultraviolet Gaussian laser beam by focus lamp and punches, the size and the step 2 of the ultraviolet gauss laser beam power that sends by control step 1) medium ultraviolet laser instrument in the punching process) middle central diameter of regulating aperture, make on the glass membrane pincers chip and form the funnel-form micropore.
In above-mentioned steps 3) in, the center spot of described ultraviolet Gaussian laser beam can be earlier by entering described focus lamp again after the mirror reflects.
In such scheme, can also adopt the ultraviolet Gaussian laser beam motionless, by moving horizontally the method for glass membrane pincers chip, finish the processing that array is arranged micropore on the whole glass membrane pincers chip.
Preferably, described glass membrane pincers chip places on the motion control platform, and the control of motion control platforms through calculation machine digital control system moves horizontally, and drive glass membrane pincers chip level moves; Computerized digital control system is also controlled the duty of ultraviolet laser.
Preferably, the wave-length coverage of the ultraviolet Gaussian laser beam that described ultraviolet laser sends is 266～355nm, and frequency range is 10～100KHz, and the pulsewidth scope is 15～50ns; The diaphragm diameter adjustable extent of described adjusting aperture is 1.2～1.8mm; Described beam expanding lens carries out the ultraviolet Gaussian laser beam 8～10 times expansion bundle.
Preferably, the entrance hole diameter of described micropore is greater than the outlet aperture, and the scope of micropore entrance hole diameter is 2～10 μ m, and the scope of micropore outlet aperture is 1～2 μ m.
Preferably, described glass membrane pincers chip thickness is 0.1mm～0.15mm; Described focus lamp is a telecentric lens, and the minimum light spot diameter after the ultraviolet Gaussian laser beam focuses on through these lens is 0.7 μ m.
Introduce laser micropore process principle of the present invention below.
1) interaction mechanism of Ultra-Violet Laser and glass inorganic material:
According to the different qualities of machined material and to the degree of absorption difference of different wavelength of laser, often select the laser of a certain specific wavelength that material is processed, in the laser micropore processing in the hope of reaching best processing effect.Consider the characteristic of glass material, it is that the ultraviolet laser of 266nm～355nm comes the micropore of glass membrane pincers chip is processed that the applicant selects the output wavelength scope.The laser of this wavelength is very suitable for inorganic material.Because the diffraction phenomena of light beam is the principal element of restriction processing component minimum dimension, the minimum light spot diameter of the accessible focus point of laser is linear increasing along with the increase of wavelength, so the laser of shorter wavelength can process littler parts.In addition, the little processing procedure of Ultra-Violet Laser is not " heat " processing procedure from essence, the high-octane photon of Ultra-Violet Laser can directly destroy the chemical bond of glass material, this process is called " photetching " effect, Ultra-Violet Laser processing is to come down to cold-treating process, heat affecting is very little, and high-octane ultraviolet light directly destroys the chemical bond of glass material, and the aperture diameter that makes this " cold " process is controlled and have a smooth uniform edge.
2) micro molding mechanism:
Be Gaussian beam on the theory of laser that laser instrument sends, desirable Gaussian beam helps the moulding of leak shape micropore shown in Figure 2, but in fact laser can not reach this perfect condition.In order to make laser as much as possible near desirable Gaussian beam pattern, the present invention combines beam expanding lens and plays the effect (as shown in Figure 3) of screening light beam with the adjusting aperture.At first, the ultraviolet gauss laser that ultraviolet laser sends expands through beam expanding lens to be restrainted, and the optical beam ratio that beam expanding lens expands behind the bundle is thicker, and this light beam has only the optical axis mid portion can pass through to regulate aperture after arriving and regulating aperture.As shown in Figure 4, size by control and regulation aperture center-hole diameter can play the effect of selecting light beam, the aperture center-hole diameter is more little to see through the laser go more near Gaussian beam, also guaranteed to obtain littler focal beam spot, the laser beam that comes out from the adjusting aperture also is a Gaussian distribution through the laser power density distribution that telecentric lens focuses on back focus, laser near ultraviolet Gaussian laser beam optical axis is higher through focusing on the back power density, the molecular structure that destroys glass material in the extremely short time forms micropore, laser power density away from ultraviolet Gaussian laser beam optical axis is much smaller, so be difficult to penetrate glass material at short notice, make the conical surface that on glass membrane pincers chip, forms micropore away from the laser of ultraviolet Gaussian laser beam optical axis.By the central diameter of control and regulation aperture, can make on the glass membrane pincers chip and form the funnel-form micropore.
The present invention adopts Ultra-Violet Laser processed glass patch clamp chips, the ultraviolet Gaussian laser beam acts on the glass membrane pincers chip after expanding bundle, regulating aperture, reflection, focusing, directly destroy the chemical bond of glass material, the scope that forms the entrance hole diameter is that the scope of 2～10 μ m, outlet aperture is the micropore of 1～2 μ m, by the diameter that the diameter of regulating aperture can be regulated the micropore of processing indirectly, regulate the tapering that glass membrane clamps the micropore that forms on the chip by the power of regulating the ultraviolet Gaussian laser beam and the central diameter of regulating aperture.Can realize the large-batch industrial processing of whole glass membrane pincers chip by Computerized digital control system and motion control platform.The present invention has opened up a new way for processing micron-sized hole and make chip carry out large-scale production on the glass membrane pincers.
Description of drawings
Fig. 1 is the fundamental diagram of single patch clamp chips;
Fig. 2 is the structural representation of patch clamp chips micropore;
Fig. 3 is the structural representation of patch clamp chips micropore processing device of the present invention;
The schematic diagram that Fig. 4 regulates for Gaussian beam;
Wherein, 1-ultraviolet laser, 2-beam expanding lens, 3-regulate aperture, 4-speculum, 5-focus lamp, 6-patch clamp chips anchor clamps, 7-motion control platform, 8-Computerized digital control system.
The specific embodiment
The present invention is described in further detail below in conjunction with drawings and Examples:
The described a kind of patch clamp chips capillary processing method of present embodiment, adopt a kind of patch clamp chips micropore processing device as shown in Figure 3, this device comprises ultraviolet laser 1, beam expanding lens 2, regulate aperture 3, speculum 4, focus lamp 5, glass membrane pincers chip fixture 6, Computerized digital control system 8 and motion control platform 7, wherein, beam expanding lens 2 is set on the output light path of ultraviolet laser 1, be provided with on the output light path of beam expanding lens 2 and regulate aperture 3, on the output light path of adjusting aperture 3 speculum 4 is set, on the reflection output light path of speculum 4 focus lamp 5 is set, glass membrane pincers chip fixture 6 is set on the output light path of focus lamp 5.The laser control signal output of Computerized digital control system 8 connects the control interface of ultraviolet laser 1, the position control signal output of Computerized digital control system 8 connects the control interface of motion control platform 7, and glass membrane pincers chip fixture 6 places on the motion control platform 7.
The laser beam expanding multiple of the beam expanding lens 2 of this device is 8～10 times; The diaphragm diameter adjustable extent of regulating aperture 3 is 1.2mm～1.8mm; The wavelength of the ultraviolet Gaussian laser beam that ultraviolet laser 1 sends is 266nm～355nm; The frequency of the ultraviolet Gaussian laser beam that ultraviolet laser 1 sends is 10KHz～100KHz, and the power of the ultraviolet Gaussian laser beam that ultraviolet laser 1 sends is 1～7 watt.
The course of work of the present invention is: glass membrane embedding chip to be processed is placed on the motion control platform, find the position in first hole to be processed by the ruddiness indication, open ultraviolet laser 1, ultraviolet laser 1 corresponding work parameter (comprising that electric current 20A, frequency are 15-20KHz) is set, make laser works under rated current and temperature, the pattern of ultraviolet laser 1 is set to MODE_1 (first pulse suppression mode), makes the laser energy of output be in more stable state.It is the center spot of the ultraviolet Gaussian laser beam of 1.2～1.8mm by regulating aperture 3 taking-up pore diameter ranges that the ultraviolet Gaussian laser beam expands Shu Houzai by beam expanding lens 2, change the angle of ultraviolet Gaussian laser beam then by speculum 4, focus on by 5 pairs of Ultra-Violet Lasers of focus lamp at last, luminous power on the glass material that arrival glass membrane pincers pincers chip fixture 6 is clamped after focusing on is between 1～7W, and on glass material, get out the micropore hole, regulate focus lamp 5 and will make the focus of ultraviolet Gaussian laser beam just drop on the upper surface of glass membrane embedding chip to be processed.By the central diameter of control and regulation aperture, making and forming the entrance hole diameter scope on the glass membrane pincers chip is 2～10 μ m, and micropore outlet aperture scope is the funnel-form micropore of 1～2 μ m.In addition, in process, adopt laser beam motionless, by Computerized digital control system 8 controlled motions control platform 7, move horizontally the method for glass membrane pincers chip, finish the processing that array is arranged micropore on the whole glass membrane pincers chip, realized the automation of whole work flow.
In addition, when present embodiment was implemented, regulating diaphragm diameter size was 1.5mm, opened laser and after 1 second of delaying time, and took off glass membrane and clamped chip fixture 6 measure pore size under light microscope; If glass material is not punched, then can't measure its size, show that laser energy is not enough, the laser energy appropriateness need be transferred greatly till can just punching, otherwise, if the light hole aperture then needs the laser energy appropriateness is turned down till meeting the demands greater than 2 μ m; To regulate the diameter of aperture if the incidence hole aperture is bigger than normal and turn down, vice versa.Adjusting test by repeatedly guarantees that finally the aperture that processes reaches till the desired value.According to the machined parameters in first hole, finish batch machining by numerical control software.
The present invention adopts the patch clamp chips of Ultra-Violet Laser processed glass material, the Ultra-Violet Laser of high-peak power destroys material at the utmost point in the short time microstructure forms evenly hole of size, on glass process micron-sized hole and make chip carry out large-scale production opened up a new way.
The content that is not described in detail in this specification belongs to this area professional and technical personnel's known prior art.
Claims (7)
1. a patch clamp chips capillary processing method is characterized in that it comprises the steps:
The power that step 1) is sent ultraviolet laser is 1～7 watt ultraviolet Gaussian laser beam, expands bundle by beam expanding lens;
Step 2) the ultraviolet Gaussian laser beam that beam expanding lens is expanded after restrainting is the center spot of the ultraviolet Gaussian laser beam of 1.2～1.8mm by regulating aperture taking-up beam diameter scope;
Step 3) focuses on glass membrane pincers chip surface with the center spot of described ultraviolet Gaussian laser beam by focus lamp and punches, the size and the step 2 of the ultraviolet gauss laser beam power that sends by control step 1) medium ultraviolet laser instrument in the punching process) middle central diameter of regulating aperture, make on the glass membrane pincers chip and form the funnel-form micropore.
2. patch clamp chips capillary processing method according to claim 1 is characterized in that: in the described step 3), the center spot of ultraviolet Gaussian laser beam is earlier by entering described focus lamp again after the mirror reflects.
3. patch clamp chips capillary processing method according to claim 1 and 2, it is characterized in that: in step 3), adopt the ultraviolet Gaussian laser beam motionless,, finish the processing that array is arranged micropore on the whole glass membrane pincers chip by moving horizontally the method for glass membrane pincers chip.
4. patch clamp chips capillary processing method according to claim 3, it is characterized in that: described glass membrane pincers chip places on the motion control platform, the control of motion control platforms through calculation machine digital control system moves horizontally, and drive glass membrane pincers chip level moves; Computerized digital control system is also controlled the duty of ultraviolet laser.
5. patch clamp chips capillary processing method according to claim 1 and 2 is characterized in that: the wave-length coverage of the ultraviolet Gaussian laser beam that described ultraviolet laser sends is 266nm～355nm, and frequency range is 10～100KHz, and the pulsewidth scope is 15～50ns; The diaphragm diameter adjustable extent of described adjusting aperture is 1.2mm～1.8mm; Described beam expanding lens carries out the ultraviolet Gaussian laser beam 8～10 times expansion bundle.
6. patch clamp chips capillary processing method according to claim 1 and 2 is characterized in that: the entrance hole diameter of described micropore is greater than the outlet aperture, and the scope of micropore entrance hole diameter is 2～10 μ m, and the scope of micropore outlet aperture is 1～2 μ m.
7. patch clamp chips capillary processing method according to claim 1 and 2 is characterized in that: glass membrane pincers chip thickness is 0.1mm～0.15mm; Described focus lamp is a telecentric lens, and the minimum light spot diameter after the ultraviolet Gaussian laser beam focuses on through these lens is 0.7 μ m.
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| Application Number | Priority Date | Filing Date | Title |
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| CN2011100347475A CN102151997A (en) | 2011-01-31 | 2011-01-31 | Method for processing micropore of patch clamp chip |
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| CN2011100347475A CN102151997A (en) | 2011-01-31 | 2011-01-31 | Method for processing micropore of patch clamp chip |
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Cited By (7)
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| CN103833202A (en) * | 2012-11-23 | 2014-06-04 | 李帅 | Portable device perforating method and apparatus thereof |
| CN104439721A (en) * | 2013-09-18 | 2015-03-25 | 大族激光科技产业集团股份有限公司 | Method for punching micron-grade-diameter hole in film through ultraviolet laser |
| CN104588871A (en) * | 2015-02-10 | 2015-05-06 | 昆山乙盛机械工业有限公司 | Adjustable laser processing device and method |
| CN106077976A (en) * | 2016-08-24 | 2016-11-09 | 江阴名鸿车顶系统有限公司 | Spoiler laser drilling system and drilling technology thereof |
| CN106624390A (en) * | 2017-03-20 | 2017-05-10 | 中国科学院高能物理研究所 | Laser drilling device |
| CN109097264A (en) * | 2018-07-31 | 2018-12-28 | 大连理工大学 | A kind of micro-current controlled cell absorption chip for membrane elastic property |
| CN111975231A (en) * | 2020-07-22 | 2020-11-24 | 广东大族粤铭激光集团股份有限公司 | Laser micropore machining method |
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Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103833202A (en) * | 2012-11-23 | 2014-06-04 | 李帅 | Portable device perforating method and apparatus thereof |
| CN104439721A (en) * | 2013-09-18 | 2015-03-25 | 大族激光科技产业集团股份有限公司 | Method for punching micron-grade-diameter hole in film through ultraviolet laser |
| CN104439721B (en) * | 2013-09-18 | 2016-05-25 | 大族激光科技产业集团股份有限公司 | Adopting the ultraviolet laser footpath of punching on film is the method in the hole of micron level |
| CN104588871A (en) * | 2015-02-10 | 2015-05-06 | 昆山乙盛机械工业有限公司 | Adjustable laser processing device and method |
| CN106077976A (en) * | 2016-08-24 | 2016-11-09 | 江阴名鸿车顶系统有限公司 | Spoiler laser drilling system and drilling technology thereof |
| CN106624390A (en) * | 2017-03-20 | 2017-05-10 | 中国科学院高能物理研究所 | Laser drilling device |
| CN109097264A (en) * | 2018-07-31 | 2018-12-28 | 大连理工大学 | A kind of micro-current controlled cell absorption chip for membrane elastic property |
| CN109097264B (en) * | 2018-07-31 | 2021-10-15 | 大连理工大学 | Microfluidic cell adsorption chip for cell patch clamp |
| CN111975231A (en) * | 2020-07-22 | 2020-11-24 | 广东大族粤铭激光集团股份有限公司 | Laser micropore machining method |
| CN111975231B (en) * | 2020-07-22 | 2022-03-18 | 广东大族粤铭激光集团股份有限公司 | Laser micropore machining method |
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Application publication date: 20110817 |