CN202093046U - Device for estimating malignant degree of neuroblastoma - Google Patents
Device for estimating malignant degree of neuroblastoma Download PDFInfo
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- CN202093046U CN202093046U CN 201020572295 CN201020572295U CN202093046U CN 202093046 U CN202093046 U CN 202093046U CN 201020572295 CN201020572295 CN 201020572295 CN 201020572295 U CN201020572295 U CN 201020572295U CN 202093046 U CN202093046 U CN 202093046U
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- control system
- hpicm
- neuroblastoma
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01Q—SCANNING-PROBE TECHNIQUES OR APPARATUS; APPLICATIONS OF SCANNING-PROBE TECHNIQUES, e.g. SCANNING PROBE MICROSCOPY [SPM]
- G01Q60/00—Particular types of SPM [Scanning Probe Microscopy] or microscopes; Essential components thereof
- G01Q60/44—SICM [Scanning Ion-Conductance Microscopy] or apparatus therefor, e.g. SICM probes
Abstract
A device for estimating malignant degree of neuroblastoma is characterized by comprising a glass microprobe full of electrolyte, an Ag/AgCI electrode, a reference Ag/AgCI electrode, a culture vessel containing cells and cell culture fluid, an external current amplifier, a digital to analog/analog to digital converter, an HPICM scanning control system, an HPICM scanning probe and a computer control system, wherein the Ag/AgCI electrode arranged in the glass microprobe is immersed in the electrolyte. The device for estimating malignant degree of the neuroblastoma has the advantages of being capable of conducting real-time detection to the cells under normal physiological conditions, visual and simple in whole diagnosis process and high in diagnosis accuracy without conducting special treatment to samples.
Description
(1) technical field:
The utility model provides a kind of device of assessing the neuroblastoma grade malignancy, especially provide and be tested and appraised the device that the relative ratios of different cell subsets in the neuroblastoma assesses its grade malignancy, in particular, be to utilize probe dancing mode scan ion electricity to lead the device that relative ratios that microscopy is tested and appraised different subtype according to the shape characteristic of different cell subsets in the neuroblastoma diagnoses this malignancy of tumor degree.
(2) background technology:
Neuroblastoma (NB) is the outer malignant solid tumor of the common cranium of a kind of child.To its never suitable diagnosis and treatment means, especially fail to diagnose early and when developing into third and fourth phase, its cure rate and prognosis effect are poorer when this disease in the past more than 20 year.The main difficulty of diagnoses and treatment neuroblastoma is heterogeneity and the tumor stem cell characteristic that comes from this tumour cell self.
Neuroblastoma cell mainly contains three kinds of cell subsets and forms: N type (neuroblast type), S type (matrix adhesive type cell) and I type (osculant cell), and these three kinds of hypotypes are the spontaneous appearance of meeting in incubation, and their biochemical characteristic, differentiation potential, grade malignancy etc. are different.On biochemical characteristic, the multipotential stem cell characteristic of I type is the most remarkable, and its renewal, competence for added value are also the strongest, and I type cell has been expressed the characteristic biochemical indicator of N type and S type simultaneously.According to grade malignancy, the I type is the strongest, the N type secondly, the S type is the most weak.From the angle of further differentiation, can also there be situation about transforming mutually in the I type to N type, the differentiation of S type between N type and the S type.The factor affecting of this many dynamic change the grade malignancy of neuroblastoma, increased the difficulty of diagnoses and treatment.
A large amount of clinical cases show: S type cell content height, the differentiation of cell height are the features of optimum neuroblastoma, and the grade malignancy of I type, N type cell and tumour and recurrence have substantial connection, therefore can identify the neuroblastoma grade malignancy by analyzing three kinds of hypotype relative ratios.At present, the most direct diagnostic means is exactly to utilize microscopy to assess the grade malignancy of knurl according to the shape characteristic of three kinds of cell subsets in the neuroblastoma.Yet ordinary optical microscope is owing to be subjected to the restriction of optical diffraction limit, and its highest resolution can not satisfy differentiates the close I type of form and the needs of N type cell; Need during Electronic Speculum is surveyed biological sample is cured and metal spraying is handled, not only waste time and energy but also can change even destroy the sample surfaces pattern, reduce the accuracy of evaluation; Owing to utilize the acting force of needle point and sample room to carry out negative feedback control, it is inevitable that atomic force microscope (AFM) probe and active somatic cell cell space slight contacts, and sharply change and during the complex biological sample (for example: circular neuroblastoma N type and I type cell) low at scanning height with substrate degree of adhering to, exist negative feedback control microscopy the difficulty that can't go beyond.
2009, the scan ion electricity of negative feedback control lead the probe dancing mode scan ion electricity that improves on microscopy (SICM) the basis on and come lead microscopy (HPICM) not only acted on original can be directly real-time, contactless under physiology liquid state condition of culture, high resolving power the advantage of detection living body biological sample surfaces three-dimensional microcosmic structure, and overcome the deficiency of the continuous negative feedback gated sweep of traditional SICM pattern, realized highly sharply changing and the nanoscale high resolution scanning imaging (see figure 2) of the living body biological sample of configuration of surface complexity.This just makes under physiological condition, contactless, high resolving power is identified the relative ratios of different cell subsets in the neuroblastoma and diagnosed its grade malignancy to become possibility.
(3) utility model content:
The utility model provides a kind of device of assessing the neuroblastoma grade malignancy, its utilization probe dancing mode scan ion electricity lead microscopy at ambient temperature (23 ± 2 ℃) to neuroblastoma cell carry out under the physiological condition, contactless, high resolution scanning imaging, and then identify the relative ratios of three kinds of cell subsets, the grade malignancy of assessment neuroblastoma.
The technical solution of the utility model: a kind of device of assessing the neuroblastoma grade malignancy is characterized in that it comprises the glass microprobe that is full of electrolytic solution, places the Ag/AgCl electrode, reference Ag/AgCl electrode, the double dish that includes cell and cell culture fluid, external current amplifier, digital-to-analogue/analog to digital converter, HPICM scanning control system, HPICM scanning head and the computer control system that are dipped in electrolytic solution in the glass microprobe; The tip of the said glass microprobe that is full of electrolytic solution and the cell culture fluid that reference Ag/AgCl electrode all places double dish; The said Ag/AgCl electrode that is dipped in electrolytic solution in the glass microprobe that places is connected with the HPICM scanning control system with external current amplifier respectively; Said reference Ag/AgCl electrode is connected with external current amplifier; Said digital-to-analogue/analog to digital converter is respectively at being connected with computer control system with external current amplifier, HPICM scanning control system; Said HPICM scanning head is made up of to dull and stereotyped piezoelectric ceramics positioning system to dull and stereotyped piezoelectric ceramics and an XY a Z, is connected with the HPICM scanning control system respectively; Said computer control system is connected respectively at external current amplifier, digital-to-analogue/analog to digital converter and HPICM scanning control system.
Above-mentioned said glass microprobe draws the instrument drawing by borosilicate microelectrode glass capillary through program control horizontal laser light microelectrode and forms borosilicate microelectrode glass capillary OD 1.00mm, ID 0.59mm; Use most advanced and sophisticated inner diameter～50nm, the glass microprobe of resistance～150M Ω in this device.
Above-mentioned said external current amplifier adopts the Multiclamp 700B amplifier of U.S. Molecular Device company; The HPICM scanning control system adopts the ICnano SICM non-contact scanning ionic conductance microscope control system of Britain Ionscope company.
Above-mentioned said digital-to-analogue/analog to digital converter adopts the Digidata1440A digital-to-analogue/analog to digital converter of U.S. Molecular Device company.
Above-mentioned said Z adopts the dull and stereotyped piezoelectric ceramics of 25 μ mLISA high precision of German Physik Instrumente company to dull and stereotyped piezoelectric ceramics, and XY is to the dull and stereotyped piezoelectric ceramics scan table of 100 * 100 μ m PIHera nanoscales of dull and stereotyped piezoelectric ceramics positioning system using Germany PhysikInstrumente company.
The principle of work of this device: by adopting ICnano SICM Control Component (the Ionscope Ltd of SBC6711DSP mainboard, Britain) control the Z that has probe to the XY of the dull and stereotyped piezoelectric ceramics scan table of nanoscale that jumps up and down and be loaded with biological sample of dull and stereotyped piezoelectric ceramics to moving, and monitor the electric current that flows into nano-dimension glass microprobe in real time with the high-speed sampling frequency of 20kHz and change; Utilize MultiClamp 700B patch clamp amplifier (Molecular Devices, the U.S.) to provide+voltage of 200mV for the microprobe of HPSICM; HPSICM no longer adopts the continuous negative feedback scan control pattern of traditional SICM, the substitute is microprobe and scans with the pattern that setting amplitude Z jumps up and down on the biological sample surface.
The course of work of the present utility model: (1) is because typical N type cell cell space is rounded, refractivity forms the neural network structure by force, each other; Typical S type cell is flat, adherent tight; And the pattern of I type cell is between N type and S type, and the individual cells shape is symmetry relatively; According to above-mentioned pattern difference, the typical three kinds of hypotype cells of picked at random utilize probe dancing mode scan ion electricity to lead microscopy it are carried out scanning imagery respectively; (2) utilize the imaging analysis software measurement to go out the height and the volume of each imaging cell, carry out statistical study by these topographic data of hypotype pair cell then, obtain the otherness between the scope of the cell height of three kinds of hypotypes in the neuroblastoma cell system and volume and mean value and the three kinds of hypotypes respectively; Statistical result showed: the cell height of three kinds of cell subsets has utmost point significant difference in the neuroblastoma, so the cell height can be used as the main foundation of identifying three kinds of hypotypes; The otherness of three kinds of hypotype cell volumes is different, so can be used as the reference frame of identifying three kinds of cell subsets; (3), accurately judge at random type under the imaging cell, and then the relative ratios who identifies three kinds of cell subsets in the neuroblastoma is used to assess the grade malignancy of this tumour according to three kinds of cell subsets standards of perfection of neuroblastoma.Imaging analysis software in the above-mentioned said step (2) adopts SICM Image Viewer analysis software.
Superiority of the present utility model is: only need the neuroblastoma cell that separates is carried out daily cultivation, need not different cell subsets are screened separation, also need not pair cell and be cured pre-service such as dyeing, can under the normal physiological conditions of tumour cell, identify in real time it, process is intuitively easy, the accuracy height of evaluation.
(4) description of drawings:
Fig. 1 is the related a kind of structural representation of assessing the device of neuroblastoma grade malignancy of the utility model.
Fig. 2 utilizes probe dancing mode scan ion electricity to lead microscope carries out scanning imagery to neuroblastoma cell principle and cell model synoptic diagram.
Fig. 3 is that (wherein, the cell volume that * represents N type and S type, N type and I type is obvious difference P＜0.005 all, n=12 for the cell height of three kinds of cell subsets in the neuroblastoma cell and volume; # represents the very obvious P of the cell difference in height of N type and S type, S type and I type, N type and I type＜0.0005, n=12).
(5) embodiment:
Embodiment: a kind of device (see figure 1) of assessing the neuroblastoma grade malignancy is characterized in that it comprises the glass microprobe that is full of electrolytic solution, places the Ag/AgCl electrode, reference Ag/AgCl electrode, the double dish that includes cell and cell culture fluid, external current amplifier, digital-to-analogue/analog to digital converter, HPICM scanning control system, HPICM scanning head and the computer control system that are dipped in electrolytic solution in the glass microprobe; The tip of the said glass microprobe that is full of electrolytic solution and the cell culture fluid that reference Ag/AgCl electrode all places double dish; The said Ag/AgCl electrode that is dipped in electrolytic solution in the glass microprobe that places is connected with the HPICM scanning control system with external current amplifier respectively; Said reference Ag/AgCl electrode is connected with external current amplifier; Said digital-to-analogue/analog to digital converter is respectively at being connected with computer control system with external current amplifier, HPICM scanning control system; Said HPICM scanning head is made up of to dull and stereotyped piezoelectric ceramics positioning system to dull and stereotyped piezoelectric ceramics and an XY a Z, is connected with the HPICM scanning control system respectively; Said computer control system is connected respectively at external current amplifier, digital-to-analogue/analog to digital converter and HPICM scanning control system.
Above-mentioned said glass microprobe draws the instrument drawing by borosilicate microelectrode glass capillary through program control horizontal laser light microelectrode and forms borosilicate microelectrode glass capillary OD 1.00mm, ID 0.59mm; Use most advanced and sophisticated inner diameter～50nm, the glass microprobe of resistance～150M Ω in this device.
Above-mentioned said external current amplifier adopts the Multiclamp 700B amplifier of U.S. Molecular Device company; The HPICM scanning control system adopts the ICnano SICM non-contact scanning ionic conductance microscope control system of Britain Ionscope company.
Above-mentioned said digital-to-analogue/analog to digital converter adopts the Digidata1440A digital-to-analogue/analog to digital converter of U.S. Molecular Device company.
Above-mentioned said Z adopts the dull and stereotyped piezoelectric ceramics of 25 μ mLISA high precision of German Physik Instrumente company to dull and stereotyped piezoelectric ceramics, and XY is to the dull and stereotyped piezoelectric ceramics scan table of 100 * 100 μ m PIHera nanoscales of dull and stereotyped piezoelectric ceramics positioning system using Germany Physik Instrumente company.
More than embodiment of the present utility model is had been described in detail, but described content only is preferred embodiment of the present utility model, can not be considered to be used to limit practical range of the present utility model; In addition, the utility model provides a kind of appraisal procedure, and the relative ratios of three kinds of cell subsets and the quantitative relationship between the grade malignancy do not belong to covering scope of the present utility model in the neuroblastoma.All equalizations of doing according to the utility model application range change and improve etc., all should still belong within the covering scope of the utility model patent.
Claims (5)
1. device of assessing the neuroblastoma grade malignancy is characterized in that it comprises the glass microprobe that is full of electrolytic solution, places the Ag/AgCl electrode, reference Ag/AgCl electrode, the double dish that includes cell and cell culture fluid, external current amplifier, digital-to-analogue/analog to digital converter, HPICM scanning control system, HPICM scanning head and the computer control system that are dipped in electrolytic solution in the glass microprobe; The tip of the said glass microprobe that is full of electrolytic solution and the cell culture fluid that reference Ag/AgCl electrode all places double dish; The said Ag/AgCl electrode that is dipped in electrolytic solution in the glass microprobe that places is connected with the HPICM scanning control system with external current amplifier respectively; Said reference Ag/AgCl electrode is connected with external current amplifier; Said digital-to-analogue/analog to digital converter is respectively at being connected with computer control system with external current amplifier, HPICM scanning control system; Said HPICM scanning head is made up of to dull and stereotyped piezoelectric ceramics positioning system to dull and stereotyped piezoelectric ceramics and an XY a Z, is connected with the HPICM scanning control system respectively; Said computer control system is connected respectively at external current amplifier, digital-to-analogue/analog to digital converter and HPICM scanning control system.
2. according to the said a kind of device of assessing the neuroblastoma grade malignancy of claim 1, it is characterized in that said glass microprobe draws the instrument drawing by borosilicate microelectrode glass capillary through program control horizontal laser light microelectrode and forms, borosilicate microelectrode glass capillary OD 1.00mm, ID 0.59mm; Use most advanced and sophisticated inner diameter～50nm, the glass microprobe of resistance～150M Ω in this device.
3. according to the said a kind of device of assessing the neuroblastoma grade malignancy of claim 1, it is characterized in that said external current amplifier adopts the Multiclamp 700B amplifier of U.S. Molecular Device company; The HPICM scanning control system adopts the ICnano SICM non-contact scanning ionic conductance microscope control system of Britain Ionscope company.
4. according to the said a kind of device of assessing the neuroblastoma grade malignancy of claim 1, it is characterized in that said digital-to-analogue/analog to digital converter adopts the Digidata1440A digital-to-analogue/analog to digital converter of U.S. Molecular Device company.
5. according to the said a kind of device of assessing the neuroblastoma grade malignancy of claim 1, it is characterized in that said Z adopts the dull and stereotyped piezoelectric ceramics of 25 μ mLISA high precision of German Physik Instrumente company to dull and stereotyped piezoelectric ceramics, XY is to the dull and stereotyped piezoelectric ceramics scan table of 100 * 100 μ m PIHera nanoscales of dull and stereotyped piezoelectric ceramics positioning system using Germany Physik Instrumente company.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 201020572295 CN202093046U (en) | 2010-10-22 | 2010-10-22 | Device for estimating malignant degree of neuroblastoma |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 201020572295 CN202093046U (en) | 2010-10-22 | 2010-10-22 | Device for estimating malignant degree of neuroblastoma |
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| CN202093046U true CN202093046U (en) | 2011-12-28 |
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| CN 201020572295 Expired - Fee Related CN202093046U (en) | 2010-10-22 | 2010-10-22 | Device for estimating malignant degree of neuroblastoma |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102455371A (en) * | 2010-10-22 | 2012-05-16 | 国家纳米技术与工程研究院 | Device and method for evaluating malignancy degree of neuroblastoma |
| CN110514634A (en) * | 2019-09-02 | 2019-11-29 | 华东理工大学 | Unicellular glycosyl metabolism labeling method based on glass nano electrode |
-
2010
- 2010-10-22 CN CN 201020572295 patent/CN202093046U/en not_active Expired - Fee Related
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102455371A (en) * | 2010-10-22 | 2012-05-16 | 国家纳米技术与工程研究院 | Device and method for evaluating malignancy degree of neuroblastoma |
| CN110514634A (en) * | 2019-09-02 | 2019-11-29 | 华东理工大学 | Unicellular glycosyl metabolism labeling method based on glass nano electrode |
| CN110514634B (en) * | 2019-09-02 | 2021-09-21 | 华东理工大学 | Single-cell glycosyl metabolism marking method based on glass nano electrode |
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| Date | Code | Title | Description |
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| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20111228 Termination date: 20141022 |
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| EXPY | Termination of patent right or utility model |