CN112051252A - Sample cell and preparation method and application thereof - Google Patents
Sample cell and preparation method and application thereof Download PDFInfo
- Publication number
- CN112051252A CN112051252A CN202010972544.XA CN202010972544A CN112051252A CN 112051252 A CN112051252 A CN 112051252A CN 202010972544 A CN202010972544 A CN 202010972544A CN 112051252 A CN112051252 A CN 112051252A
- Authority
- CN
- China
- Prior art keywords
- sample
- transparent substrate
- sealing cover
- sample cell
- sealed cavity
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 238000002360 preparation method Methods 0.000 title abstract description 9
- 239000000758 substrate Substances 0.000 claims abstract description 46
- 238000007789 sealing Methods 0.000 claims abstract description 42
- 230000005684 electric field Effects 0.000 claims abstract description 33
- 238000003384 imaging method Methods 0.000 claims abstract description 24
- 238000001917 fluorescence detection Methods 0.000 claims abstract description 18
- 239000010408 film Substances 0.000 claims description 57
- 238000000034 method Methods 0.000 claims description 14
- 239000010409 thin film Substances 0.000 claims description 14
- 239000000853 adhesive Substances 0.000 claims description 8
- 230000001070 adhesive effect Effects 0.000 claims description 8
- 238000005070 sampling Methods 0.000 claims description 7
- 238000005286 illumination Methods 0.000 claims description 6
- BQCADISMDOOEFD-UHFFFAOYSA-N Silver Chemical compound [Ag] BQCADISMDOOEFD-UHFFFAOYSA-N 0.000 claims description 4
- 239000011521 glass Substances 0.000 claims description 3
- 239000011810 insulating material Substances 0.000 claims description 3
- 239000010453 quartz Substances 0.000 claims description 3
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N silicon dioxide Inorganic materials O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims description 3
- 238000002073 fluorescence micrograph Methods 0.000 claims description 2
- 210000001520 comb Anatomy 0.000 claims 1
- 239000000523 sample Substances 0.000 description 81
- 239000000463 material Substances 0.000 description 8
- 239000002245 particle Substances 0.000 description 7
- 230000033001 locomotion Effects 0.000 description 5
- 230000008569 process Effects 0.000 description 5
- 230000003287 optical effect Effects 0.000 description 4
- 238000004720 dielectrophoresis Methods 0.000 description 3
- 238000005516 engineering process Methods 0.000 description 3
- 238000005424 photoluminescence Methods 0.000 description 3
- 230000007704 transition Effects 0.000 description 3
- 238000007738 vacuum evaporation Methods 0.000 description 3
- 239000004698 Polyethylene Substances 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- 238000009792 diffusion process Methods 0.000 description 2
- 239000003814 drug Substances 0.000 description 2
- 238000005566 electron beam evaporation Methods 0.000 description 2
- 239000007850 fluorescent dye Substances 0.000 description 2
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 description 2
- 239000010931 gold Substances 0.000 description 2
- 229910052737 gold Inorganic materials 0.000 description 2
- 238000011065 in-situ storage Methods 0.000 description 2
- 238000001755 magnetron sputter deposition Methods 0.000 description 2
- 239000000155 melt Substances 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 229920000573 polyethylene Polymers 0.000 description 2
- 238000007639 printing Methods 0.000 description 2
- 230000005476 size effect Effects 0.000 description 2
- 238000004528 spin coating Methods 0.000 description 2
- -1 Polyethylene Polymers 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 230000006978 adaptation Effects 0.000 description 1
- 230000003044 adaptive effect Effects 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 239000012620 biological material Substances 0.000 description 1
- 230000005540 biological transmission Effects 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 238000012512 characterization method Methods 0.000 description 1
- 239000002801 charged material Substances 0.000 description 1
- 238000003486 chemical etching Methods 0.000 description 1
- 229920001940 conductive polymer Polymers 0.000 description 1
- 239000004020 conductor Substances 0.000 description 1
- 238000007599 discharging Methods 0.000 description 1
- 238000005401 electroluminescence Methods 0.000 description 1
- 238000001962 electrophoresis Methods 0.000 description 1
- 238000005530 etching Methods 0.000 description 1
- 230000005284 excitation Effects 0.000 description 1
- 230000005281 excited state Effects 0.000 description 1
- 230000005669 field effect Effects 0.000 description 1
- 238000000799 fluorescence microscopy Methods 0.000 description 1
- 238000002189 fluorescence spectrum Methods 0.000 description 1
- 230000005283 ground state Effects 0.000 description 1
- 230000003993 interaction Effects 0.000 description 1
- 239000007791 liquid phase Substances 0.000 description 1
- 238000004020 luminiscence type Methods 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 229910052751 metal Inorganic materials 0.000 description 1
- 239000002184 metal Substances 0.000 description 1
- 229910044991 metal oxide Inorganic materials 0.000 description 1
- 150000004706 metal oxides Chemical class 0.000 description 1
- 239000011859 microparticle Substances 0.000 description 1
- 239000002086 nanomaterial Substances 0.000 description 1
- 239000002105 nanoparticle Substances 0.000 description 1
- 230000007935 neutral effect Effects 0.000 description 1
- 238000001259 photo etching Methods 0.000 description 1
- 238000006862 quantum yield reaction Methods 0.000 description 1
- 238000001228 spectrum Methods 0.000 description 1
Images
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/645—Specially adapted constructive features of fluorimeters
- G01N21/6456—Spatial resolved fluorescence measurements; Imaging
- G01N21/6458—Fluorescence microscopy
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/645—Specially adapted constructive features of fluorimeters
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/6486—Measuring fluorescence of biological material, e.g. DNA, RNA, cells
-
- H—ELECTRICITY
- H01—ELECTRIC ELEMENTS
- H01J—ELECTRIC DISCHARGE TUBES OR DISCHARGE LAMPS
- H01J37/00—Discharge tubes with provision for introducing objects or material to be exposed to the discharge, e.g. for the purpose of examination or processing thereof
- H01J37/02—Details
- H01J37/20—Means for supporting or positioning the object or the material; Means for adjusting diaphragms or lenses associated with the support
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/645—Specially adapted constructive features of fluorimeters
- G01N2021/6463—Optics
- G01N2021/6469—Cavity, e.g. ellipsoid
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
- G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
- G01N21/64—Fluorescence; Phosphorescence
- G01N21/645—Specially adapted constructive features of fluorimeters
- G01N2021/6482—Sample cells, cuvettes
Landscapes
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Analytical Chemistry (AREA)
- Biochemistry (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Physics & Mathematics (AREA)
- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Molecular Biology (AREA)
- Biomedical Technology (AREA)
- Engineering & Computer Science (AREA)
- Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)
Abstract
A sample cell and a preparation method and application thereof belong to the technical field of sample containing devices. The sample cell comprises a transparent substrate, a sealing cover, a sample inlet pipe, a sample outlet pipe and N electric field units; the sealing cover is fixed on the upper surface of the transparent substrate and forms a sealed cavity with the transparent substrate, and a sample inlet hole and a sample outlet hole which are communicated with the sealed cavity are formed in the sealing cover; the electric field unit comprises a conductive film group, a power supply device and two wires, wherein the conductive film group comprises two discrete conductive films, the two discrete conductive films are arranged on a transparent substrate in the sealed cavity, one end of each conductive film extends out of the sealed cavity, one ends of the two wires are respectively connected with the parts of the two conductive films extending out of the sealed cavity, and the other ends of the two wires are respectively connected with the anode and the cathode of the power supply device. The sample cell is suitable for various microscopic imaging devices and fluorescence detection devices, and can be used for carrying out microscopic imaging and fluorescence detection on a sample under an electric field.
Description
Technical Field
The invention belongs to the technical field of sample accommodating devices, particularly relates to a sample cell and a preparation method and application thereof, and particularly relates to application of the sample cell in microscopic imaging and fluorescence detection equipment.
Background
Electric fields are a common method of manipulating nanomaterials and biomaterials. In a liquid phase system, the material has two movement modes under the action of an electric field: one is electrophoresis, i.e. charged material moves in the direction of the electric field; the other is dielectrophoresis, i.e. the movement of neutral polarised particles along a gradient of electric field strength. Dielectrophoresis has become a widely used tool for separating, sorting, capturing and manipulating cells, microparticles, nanoparticles and biomolecules in solution. Dielectric electrophoretic materials have been widely used in the fields of biology, medicine, and material engineering. For dielectrophoresis, an important physical quantity is the polarizability at the level of a single molecule, single particle. However, at present, no means can realize quantitative characterization of polarizability of single particles, and the research on size effect of the single particles is only in a theoretical stage. Therefore, in-situ observation techniques and devices need to be developed to explore the microscopic mechanism of the electrophoretic motion size effect.
The fluorescence microscope is an optical microscope. The method utilizes the characteristic that fluorescent molecules emit fluorescence in an excited way, uses short-wavelength light to irradiate a detected object dyed by fluorescent materials so as to generate long-wavelength fluorescence after the detected object is excited, then uses an optical filter to separate the fluorescence, and qualitatively and quantitatively analyzes sample information according to the characteristic of a fluorescence signal. Compared with an optical microscope, a fluorescence microscope is an in-situ observation technology with high signal to noise ratio, and is widely applied to the fields of biology, medicine and the like. However, the fluorescent molecules are excited by the laser to emit light, and the photoluminescence process and the non-radiative transition process compete with each other, and the quantum yield of the photoluminescence is affected by the non-radiative transition process. It is known that the applied electric field can have a serious influence on the processes of photon absorption, electron transition, and electron return from the excited state to the ground state. The emission characteristics and kinetics of fluorescent molecules in biological systems are influenced by strong electric fields due to the presence of charged and polar groups in the structure of the biomolecule, which strongly influence the luminescence process of the embedded fluorescent molecule. Therefore, the evolution trend of the spectrum, emission intensity and service life of the fluorescent molecule along with the change of the external electric field intensity is researched, and not only the solution information is reflected, but also the information of the influence of the field effect on the fluorescence excitation kinetics can be provided.
However, the prior art lacks a sample cell adapted to be loaded with an electric field for microscopic imaging and fluorescence testing. A sample holding device for microscopic imaging and fluorescence detection is adaptable to a variety of microscopic and fluorescence detection equipment, including but not limited to, far field fluorescence microscopes, bright field optical microscopes, dark field microscopes, and confocal scanning fluorescence microscopes. By designing a sample cell capable of loading an adjustable electric field and applying various microscopic equipment to research the problems of (fluorescence) imaging, ultrafast fluorescence spectrum analysis, diffusion dynamics and the like of charged particles in a solution and a biological system under the electric field, the application fields and the ranges of the microscopic technology and the fluorescence detection technology can be further expanded.
Disclosure of Invention
The invention aims to solve the technical problem that an adaptive sample cell capable of loading an electric field for microscopic imaging and fluorescence testing is lacked in the prior art, and provides a sample cell and a preparation method and application thereof.
The technical scheme adopted by the invention for solving the technical problems is as follows.
The invention provides a sample cell, which comprises a transparent substrate, a sealing cover, a sample inlet pipe, a sample outlet pipe and N electric field units, wherein N is more than or equal to 1;
the sealing cover is fixed on the upper surface of the transparent substrate and forms a sealed cavity with the transparent substrate, a sample inlet hole and a sample outlet hole which are communicated with the sealed cavity are formed in the sealing cover, and the sealing cover is made of an insulating material;
the sampling pipe is connected with the sampling hole;
the sample outlet pipe is connected with the sample outlet hole;
the electric field unit consists of a conductive film group, a power supply device and a lead group, each conductive film group consists of two discrete conductive films, the two discrete conductive films are arranged on the transparent substrate in the sealed cavity, and one end of each conductive film extends out of the sealed cavity; the lead group consists of two leads, one ends of the two leads are respectively connected with the parts of the two conductive films extending out of the sealed cavity body, and the other ends of the two leads are respectively connected with the anode and the cathode of the power supply device.
Further, the transparent substrate is a glass slide or a quartz slide.
Further, the thickness of the transparent substrate is 0.1-0.17 mm.
Further, the sealing cover is of a flat plate structure, and the outer edge of the lower surface of the sealing cover is adhered and fixed on the upper surface of the transparent substrate through an insulating adhesive.
Further, the sealing cover is of a square or cylinder structure with an opening at the bottom end, and the edge of the opening at the bottom end of the sealing cover is fixedly adhered to the upper surface of the transparent substrate through an insulating adhesive.
Furthermore, the sampling pipe and the sample outlet pipe are flexible pipes.
Further, the two discrete conductive films are in the shape of a symmetrical rectangle, an interdigitated comb or a double helix.
Furthermore, the thickness of the conductive film is 0.05-2 μm.
Furthermore, the connecting part of the lead and the conductive film is fixed through conductive silver paste.
Furthermore, N conductive film groups in the N electric field units share M power supply devices, and M is less than or equal to N.
The invention also provides a preparation method of the sample cell, which comprises the following steps:
step one, preparing a conductive film group on a transparent substrate;
step two, the sealing cover is fixedly bonded on the transparent substrate;
connecting the sample inlet pipe with the sample inlet hole, and connecting the sample outlet pipe with the sample outlet hole;
and step four, assembling the conductive thin film group, the power supply device and the two leads in each electric field unit to obtain the sample cell.
Further, the conductive film is prepared by magnetron sputtering, electron beam evaporation, vacuum evaporation, spin coating or printing.
The sample cell can be applied as a sample cell of a microscopic imaging device or a fluorescence detection device.
Further, a sample to be detected is injected into the sealed cavity through the sample inlet pipe, the sample cell is placed on a sample table of the microscopic imaging equipment or the fluorescence detection equipment, the microscope is adjusted to focus on the sample to be detected, the power supply device and the lighting light path are started, the sample to be detected emits a fluorescence signal after being excited, and the fluorescence signal is collected by the imaging light path and output to the signal processor to generate a single-molecule fluorescence image or video after passing through a lens of the microscope.
Compared with the prior art, the invention has the beneficial effects that:
the sample cell provided by the invention is suitable for various microscopic imaging devices and fluorescence detection devices, and can be used for carrying out microscopic imaging and fluorescence detection on a sample under an electric field.
Drawings
In order to more clearly illustrate the technical solutions in the embodiments of the present invention, the drawings needed to be used in the embodiments will be briefly described below, and it is obvious that the drawings in the following description are only some embodiments of the present invention, and it is obvious for those skilled in the art to obtain other drawings without creative efforts.
FIG. 1 is a schematic cross-sectional view of a sample cell according to the present invention;
fig. 2 is a top view of a sample cell provided by the present invention (N-6, two discrete conductive films are shaped as symmetrical rectangles);
fig. 3 is a top view of a sample cell provided by the present invention (N ═ 1, two discrete conductive films are interdigitated in shape);
fig. 4 is a top view of a sample cell provided by the present invention (N ═ 1, two discrete conductive films are shaped as a double helix);
FIG. 5 is a schematic view of a sample cell provided by the present invention;
in the figure, 1, a transparent substrate, 2, a conductive film, 3, a sealing cover, 31, a sample inlet hole, 32, a sample outlet hole, 4, an insulating adhesive, 5, a sample inlet tube, 6, a sample outlet tube, 7, a sealing cavity, 8, conductive silver paste, 9, a lead, 10, a power supply device, 11, a microscope lens, 12, an illumination light path, 13 and an imaging light path.
Detailed Description
For a further understanding of the invention, preferred embodiments of the invention are described below in conjunction with the detailed description, but it is to be understood that the description is intended to further illustrate the features and advantages of the invention and not to limit the claims to the invention.
As shown in FIG. 1, the sample cell of the present invention comprises a transparent substrate 1, a sealing cover 3, a sample inlet tube 5, a sample outlet tube 6 and an electric field unit.
Among them, the transparent substrate 1 is a commonly used technique. The shape of the transparent substrate 1 is not particularly limited, and is generally square or circular. The material of the transparent substrate 1 may be glass, quartz, or the like. The thickness of the transparent substrate 1 is preferably 0.1-0.17 mm.
The sealing cover 3 is fixed on the upper surface of the transparent substrate 1, and forms a sealed cavity 7 with the transparent substrate 1. The shape of the sealing lid 3 is not particularly limited, and a sealed cavity can be formed with the transparent substrate 1. If it is preferable that the sealing cover 3 is a flat plate structure, the outer edge of the lower surface of the sealing cover 3 is adhesively fixed on the upper surface of the transparent substrate 1 by the insulating adhesive 4; the sealing cover 3 is preferably of a square or cylindrical structure with an open bottom end, and the edge of the open bottom end of the sealing cover 3 is adhesively fixed on the upper surface of the transparent substrate 1 by an insulating adhesive 4. The sealing cover 3 is provided with a sample inlet hole 31 and a sample outlet hole 32 which are communicated with the sealing cavity 7, preferably, the sample inlet hole 31 and the sample outlet hole 32 are arranged on a straight line which is farthest away on the upper surface of the sealing cover 3, if the upper surface of the sealing cover 3 is square, the sample inlet hole 31 and the sample outlet hole 32 are arranged at two ends of a diagonal line, and if the upper surface of the sealing cover 3 is round, the sample inlet hole 31 and the sample outlet hole 32 are arranged at two ends of a diameter. The material of the sealing cover 3 is an insulating material, preferably Polyethylene (PE).
The sample inlet pipe 5 is connected with the sample inlet hole 31 and used for conveying the solution sample or gas into the sealed cavity 7, and the sample outlet pipe 6 is connected with the sample outlet hole 32 and used for discharging the solution sample or gas in the sealed cavity 7. The sampling pipe 5 and the sampling pipe 6 are both flexible pipes.
The number of the electric field units is N, and N is more than or equal to 1. Each electric field unit consists of a conductive film group, a power supply device 10 and two lead wires 9. Each conductive film group consists of two discrete conductive films 2, the two discrete conductive films 2 are fixed on the transparent substrate 1 in the sealed cavity 7, and one end of each conductive film 2 extends to the outside of the sealed cavity 7 respectively, namely is exposed out of the sealed cavity 7 to form a connecting electrode. Typically, the conductive film 2 extending out of the sealed cavity 7 is also fixed on the transparent substrate 1. As shown in fig. 2, N is 6, each conductive thin film group is composed of two discrete conductive thin films 2, each of the two discrete conductive thin films 2 is rectangular, and is symmetrically disposed on the left and right sides of the upper surface of the transparent substrate 1, and the left end of the left conductive thin film 2 and the right end of the right conductive thin film 2 extend out of the sealed cavity 7 to form a connection electrode. As shown in fig. 3, N ═ 1, each conductive film group is composed of two discrete conductive films 2, the two discrete conductive films 2 are in the shape of an interdigitated comb, and the outer ends of the two discrete conductive films 2 extend out of the sealed cavity 7 to form connection electrodes. As shown in fig. 4, N ═ 1, each conductive film group is composed of two discrete conductive films 2, the two discrete conductive films 2 are double-helix shaped, and the outer ends of the two discrete conductive films 2 extend out of the sealed cavity 7 to form connection electrodes. The conductive film 2 is a conductive material such as a metal film, a metal oxide film, a conductive polymer, or the like. The thickness of the conductive thin film 2 is preferably 0.05 to 2 μm. The commonly used material of the conductive film 2 is gold with a thickness of 200 nm. One ends of the two leads 9 are respectively connected with the parts of the two conductive films 2 extending out of the sealed cavity 7, the joint is fixed by conductive silver paste 8, and the other ends of the two leads 9 are respectively connected with the anode and the cathode of the power supply device 10. N conductive film groups in the N electric field units share M power supply devices 10, and M is less than or equal to N; for example, if there are five electric field units, there are five groups of conductive thin films, and two groups of conductive thin films may share one power supply device 10, and the other three groups of conductive thin films each share one power supply device 10; or five groups of conductive thin film groups can share one power supply device 10; it is also possible that two sets of conductive films share one power supply device 10, and the other three sets of conductive films share one power supply device 10.
In the technical scheme, the shapes and thicknesses of the transparent substrate 1 and the sealing cover 3 are adjusted by taking the adaptation of microscopic imaging equipment or fluorescence detection equipment as a reference, and the thickness and the pattern structure of the conductive film 2 are realized by adjusting the preparation process according to the detection requirements.
The invention also provides a preparation method of the sample cell, which comprises the following steps:
step one, preparing a conductive film group on a transparent substrate 1;
step two, the sealing cover 3 is fixedly bonded on the transparent substrate 1;
thirdly, connecting the sample inlet pipe 5 with the sample inlet hole, and connecting the sample outlet pipe 6 with the sample outlet hole;
and step four, assembling the conductive thin film group, the power supply device 10 and the two leads 9 in each electric field unit to obtain the sample cell.
In the above technical solution, the preparation of the conductive film 2 is the prior art, and the methods are generally two: one is to directly prepare the conductive film 2 with the required pattern on the transparent substrate 1 by magnetron sputtering, electron beam evaporation, vacuum evaporation, spin coating or printing, etc.; the other method is to firstly prepare a conductive film covering the upper surface of the transparent substrate 1 on the transparent substrate 1 by adopting the method, and then pattern the conductive film 2 by utilizing a mask, wherein the method can be photoetching, chemical etching or AFM probe etching and the like. The selection is made in particular depending on the material of the conductive film 2. If a gold thin film is used, it is generally prepared directly on the transparent substrate 1 by vacuum evaporation.
As shown in fig. 5, the sample cell of the present invention can be applied as a sample cell of a microscopic imaging apparatus and a fluorescence detection apparatus. Firstly, a solution with fluorescent molecules, a solution with fluorescent probes, a melt with fluorescent molecules, a melt with fluorescent probes and other samples to be detected, which need to be tracked, are injected into a sealed cavity 7 through a sample inlet pipe 5, then a sample cell is placed on a sample stage of detection equipment, a microscope is adjusted to focus on the position of the sample to be detected, the output voltage or current of a power supply device 10 is set, the power supply device 10 is started (the power supply device 10 provides a transverse electric field for the sealed cavity 7 where the sample to be detected is located) and an illumination light path 12 is started (the sequence of starting the power supply device 10 and the illumination light path 12 is not limited), the fluorescent molecules in the sample to be detected emit fluorescent signals after being excited, and the fluorescent signals are collected by an imaging light path 13 after passing through a microscope lens 11 and output to a signal processor to generate a single-molecule. When the tracked fluorescent molecules or probes are charged, the fluorescent characteristics (fluorescence intensity, fluorescence lifetime, etc.) and the motion characteristics (diffusion speed, motion trajectory, etc.) of the tracked fluorescent molecules or probes are changed under an applied electric field, and the interaction of charged particles is shown in the generation of a single-molecule fluorescent tracking image or video. The output voltage or current of the power supply device 10 can be adjusted to adjust the magnitude of the electric field of the sample to be tested, and the influence of the electric field on the charged particles can be observed by changing the horizontal testing position along the gradient of the conductive film 2.
In the above technical solution, the microscopic imaging device and the fluorescence detection device are generally a microscopic fluorescence imaging apparatus, but are not limited thereto. When the sample cell of the invention is applied as a sample cell of a microscopic imaging device and a fluorescence detection device, the microscopic imaging device and the fluorescence detection device are not limited to the incident wavelength and the type of a fluorescence label, are not limited to the relative positions of an illumination light path and an imaging light path, and are not limited to a transmission and reflection working mode. The power supply device 10 may be turned on to realize the electroluminescence emission of the sample to be measured instead of the photoluminescence emission of the illumination light path 10.
It should be understood that the above embodiments are only examples for clearly illustrating the present invention, and are not intended to limit the present invention. Other variations and modifications will be apparent to persons skilled in the art in light of the above description. And are neither required nor exhaustive of all embodiments. And obvious variations or modifications of the invention may be made without departing from the spirit or scope of the invention.
Claims (10)
1. The sample cell is characterized by comprising a transparent substrate (1), a sealing cover (3), a sample inlet pipe (5), a sample outlet pipe (6) and N electric field units, wherein N is more than or equal to 1;
the sealing cover (3) is fixed on the upper surface of the transparent substrate (1) and forms a sealing cavity (7) with the transparent substrate (1), a sample inlet hole (31) and a sample outlet hole (32) which are communicated with the sealing cavity (7) are formed in the sealing cover (3), and the sealing cover (3) is made of an insulating material;
the sampling pipe (5) is connected with the sampling hole (31);
the sample outlet pipe (6) is connected with the sample outlet hole (32);
the electric field unit consists of a conductive film group, a power supply device (10) and a lead group, the conductive film group consists of two discrete conductive films, the two discrete conductive films (2) are both arranged on the transparent substrate (1) in the sealed cavity (7), and one end of each conductive film extends out of the sealed cavity (7); the lead group consists of two leads (9), one ends of the two leads (9) are respectively connected with the parts of the two conductive films (2) extending out of the sealed cavity (7), and the other ends are respectively connected with the anode and the cathode of the power supply device (10).
2. A sample cell according to claim 1, characterized in that the transparent substrate (1) is a glass or quartz plate, the transparent substrate (1) having a thickness of 0.1-0.17 mm.
3. The sample cell of claim 1,
the sealing cover (3) is of a flat plate structure, and the outer edge of the lower surface of the sealing cover (3) is adhered and fixed on the upper surface of the transparent substrate (1) through an insulating adhesive (4);
or the sealing cover (3) is of a square structure with an opening at the bottom end, and the edge of the opening at the bottom end of the sealing cover (3) is adhered and fixed on the upper surface of the transparent substrate (1) through an insulating adhesive (4);
or the sealing cover (3) is of a cylindrical structure with an opening at the bottom end, and the edge of the opening at the bottom end of the sealing cover (3) is adhered and fixed on the upper surface of the transparent substrate (1) through an insulating adhesive (4).
4. A cuvette according to claim 1, characterized in that the inlet tube (5) and the outlet tube (6) are both flexible tubes.
5. A cuvette according to claim 1, characterized in that the two discrete conductive films (2) are shaped as symmetrical rectangles, interdigitated combs or double spirals, the conductive films (2) each having a thickness of 0.05-2 μm.
6. The cuvette according to claim 1, characterized in that the connection of the lead (9) and the conductive film (2) is fixed by a conductive silver paste (8).
7. A sample cell according to claim 1, wherein N groups of conductive films in N electric field units share M power supply means (10), M ≦ N.
8. A method of preparing a sample cell according to any of claims 1 to 7, comprising the steps of:
step one, preparing a conductive film group on a transparent substrate (1);
step two, adhering and fixing the sealing cover (3) on the transparent substrate (1);
connecting the sample inlet pipe (5) with the sample inlet hole, and connecting the sample outlet pipe (6) with the sample outlet hole;
and step four, assembling the conductive thin film group, the power supply device (10) and the two leads (9) in each electric field unit to obtain the sample cell.
9. The sample cell of any of claims 1-7 can be used as a sample cell for a microscopic imaging device or a fluorescence detection device.
10. The sample cell according to any one of claims 1 to 7 can be used as a sample cell of a microscopic imaging device or a fluorescence detection device, wherein a sample to be detected is injected into the sealed cavity (7) through the sample inlet tube (5), the sample cell is placed on a sample stage of the microscopic imaging device or the fluorescence detection device, a microscope is adjusted to be focused on the sample to be detected, the power supply device (10) and the illumination light path (12) are started, the sample to be detected emits a fluorescence signal after being excited, and the fluorescence signal is collected by the imaging light path (13) after passing through a lens (11) of the microscope and is output to the signal processor to generate a single-molecule fluorescence image or video.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010972544.XA CN112051252A (en) | 2020-09-16 | 2020-09-16 | Sample cell and preparation method and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010972544.XA CN112051252A (en) | 2020-09-16 | 2020-09-16 | Sample cell and preparation method and application thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN112051252A true CN112051252A (en) | 2020-12-08 |
Family
ID=73604294
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202010972544.XA Pending CN112051252A (en) | 2020-09-16 | 2020-09-16 | Sample cell and preparation method and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN112051252A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114112920A (en) * | 2021-12-10 | 2022-03-01 | 中国科学院长春应用化学研究所 | Fluorescence detection sample pool capable of intelligently and accurately regulating and controlling solution concentration on line and control method |
Citations (14)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP1324029A2 (en) * | 2001-12-26 | 2003-07-02 | Nec Corporation | Fluorescent transilluminator |
| JP2004243193A (en) * | 2003-02-13 | 2004-09-02 | Pentax Corp | Labo-on-a-chip, method for manufacturing the sane and apparatus for the same |
| JP2005069893A (en) * | 2003-08-25 | 2005-03-17 | Shimadzu Corp | Surface plasmon resonance device and analyzer using the same |
| WO2007046485A1 (en) * | 2005-10-19 | 2007-04-26 | Sharp Kabushiki Kaisha | Electrophoretic chip, electrophoretic device, and electrophoretic system |
| JP2008196860A (en) * | 2007-02-08 | 2008-08-28 | Ario Techno Kk | Particulate detection device and microorganism detection method |
| CN101343613A (en) * | 2008-08-22 | 2009-01-14 | 重庆大学 | Flexible high-pass cell electric amalgamation microelectrode array chip apparatus |
| CN101460253A (en) * | 2006-05-31 | 2009-06-17 | 明达西德实验室有限公司 | Method and apparatus for manipulating single cells and small aggregates thereof |
| CN101693875A (en) * | 2009-09-30 | 2010-04-14 | 重庆大学 | Cell electrofusion chip device based on columnar microelectrode array and electrofusion method |
| CN201477074U (en) * | 2009-09-04 | 2010-05-19 | 孙朝阳 | Single-plate lateral order electrophresis apparatus |
| CN102302898A (en) * | 2011-06-17 | 2012-01-04 | 西安交通大学 | Micro blood cell separation device and method for using same |
| CN105358971A (en) * | 2013-07-01 | 2016-02-24 | 莱卡微系统Cms有限责任公司 | Laser microdissection system and examination method for samples containing nucleic acid |
| CN105572046A (en) * | 2014-10-16 | 2016-05-11 | 中国科学院物理研究所 | Fluorescence detection sample pool and preparation method thereof |
| CN107615041A (en) * | 2015-10-07 | 2018-01-19 | Afi技术公司 | Check device, inspection system and inspection method |
| WO2018078999A1 (en) * | 2016-10-27 | 2018-05-03 | シャープ株式会社 | Fluorescent testing system, dielectrophoresis device, and molecular testing method |
-
2020
- 2020-09-16 CN CN202010972544.XA patent/CN112051252A/en active Pending
Patent Citations (14)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP1324029A2 (en) * | 2001-12-26 | 2003-07-02 | Nec Corporation | Fluorescent transilluminator |
| JP2004243193A (en) * | 2003-02-13 | 2004-09-02 | Pentax Corp | Labo-on-a-chip, method for manufacturing the sane and apparatus for the same |
| JP2005069893A (en) * | 2003-08-25 | 2005-03-17 | Shimadzu Corp | Surface plasmon resonance device and analyzer using the same |
| WO2007046485A1 (en) * | 2005-10-19 | 2007-04-26 | Sharp Kabushiki Kaisha | Electrophoretic chip, electrophoretic device, and electrophoretic system |
| CN101460253A (en) * | 2006-05-31 | 2009-06-17 | 明达西德实验室有限公司 | Method and apparatus for manipulating single cells and small aggregates thereof |
| JP2008196860A (en) * | 2007-02-08 | 2008-08-28 | Ario Techno Kk | Particulate detection device and microorganism detection method |
| CN101343613A (en) * | 2008-08-22 | 2009-01-14 | 重庆大学 | Flexible high-pass cell electric amalgamation microelectrode array chip apparatus |
| CN201477074U (en) * | 2009-09-04 | 2010-05-19 | 孙朝阳 | Single-plate lateral order electrophresis apparatus |
| CN101693875A (en) * | 2009-09-30 | 2010-04-14 | 重庆大学 | Cell electrofusion chip device based on columnar microelectrode array and electrofusion method |
| CN102302898A (en) * | 2011-06-17 | 2012-01-04 | 西安交通大学 | Micro blood cell separation device and method for using same |
| CN105358971A (en) * | 2013-07-01 | 2016-02-24 | 莱卡微系统Cms有限责任公司 | Laser microdissection system and examination method for samples containing nucleic acid |
| CN105572046A (en) * | 2014-10-16 | 2016-05-11 | 中国科学院物理研究所 | Fluorescence detection sample pool and preparation method thereof |
| CN107615041A (en) * | 2015-10-07 | 2018-01-19 | Afi技术公司 | Check device, inspection system and inspection method |
| WO2018078999A1 (en) * | 2016-10-27 | 2018-05-03 | シャープ株式会社 | Fluorescent testing system, dielectrophoresis device, and molecular testing method |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114112920A (en) * | 2021-12-10 | 2022-03-01 | 中国科学院长春应用化学研究所 | Fluorescence detection sample pool capable of intelligently and accurately regulating and controlling solution concentration on line and control method |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| US10900999B2 (en) | Microscopy imaging | |
| CN106092865B (en) | It is a kind of based on digital microcurrent-controlled fluorescence drop separation system and its method for separating | |
| US7615762B2 (en) | Method and apparatus for low quantity detection of bioparticles in small sample volumes | |
| JP2832117B2 (en) | Sample measuring device and sample measuring system | |
| US20100243888A1 (en) | Apparatus and Method for Inspecting Samples | |
| KR101592241B1 (en) | Method of fabricating a device having an array of nano particles, surface plasmon based sensor, and method of assaying substances using the sensor | |
| US20040058423A1 (en) | Fast electrical lysis of cells and rapid collection of the contents thereof using capillary electrophoresis | |
| EP2458383B1 (en) | Method of electrochemically detecting a sample substance | |
| EA001198B1 (en) | Multi-array, multi-specific electrochemiluminescence testing | |
| EP2048429A1 (en) | Illuminating device, illuminating method, light detector and light detecting method | |
| Chen et al. | Surface plasmon resonance (SPR) combined technology: A powerful tool for investigating interface phenomena | |
| CN110646493A (en) | Microfluidic chip, protein detection method, device and system | |
| CN112051252A (en) | Sample cell and preparation method and application thereof | |
| CN108507991B (en) | Two-photon fluorescence enhancement method and application thereof | |
| Marchand et al. | Optical near-field electron microscopy | |
| CN114724921A (en) | Automatic electrospray spraying device and method for mass spectrometry imaging sample preparation | |
| Tan et al. | Honeycomb meshed working electrodes based on microsphere lithography for high-resolution chemical image sensor | |
| US9081005B2 (en) | Biochip | |
| EP4336170A1 (en) | Method for accumulating minute object, and method for detecting minute object using same | |
| CN112986171B (en) | Plasmon resonance enhanced substrate and preparation method and application thereof | |
| Wang et al. | Photoelectrochemical detection of Cd 2+ based on in situ electrodeposition of CdS on ZnO nanorods | |
| KR20150117198A (en) | Sensor for spectroscopic analysis | |
| CN104880453B (en) | The photoelectric synchronous method for sensing of solid nano passage based on dark-field imaging | |
| KR101759894B1 (en) | Lab-on-a-chip and a method of fabricating thereof | |
| CN115855930A (en) | Novel method for solar cell photocurrent imaging |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20201208 |
|
| WD01 | Invention patent application deemed withdrawn after publication |