CN105717100A - Detecting system capable of accurately acquiring ECL (electrochemiluminescence) spectral information - Google Patents
Detecting system capable of accurately acquiring ECL (electrochemiluminescence) spectral information Download PDFInfo
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- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/75—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated
- G01N21/76—Chemiluminescence; Bioluminescence
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Abstract
The invention relates to a detecting system capable of accurately acquiring ECL (electrochemiluminescence) spectral information. The detecting system comprises an electrochemistry equipment unit, an ECL radiation generation device unit, a spectrum detection device unit and a data processing and control unit, wherein the ECL radiation generation device unit comprises an ECL cell as well as a working electrode, a reference electrode and a counter electrode which are connected with the electrochemistry equipment unit; the spectrum detection device unit comprises a collection objective lens and a CCD grating spectrometer; the ECL radiation generation device unit and the collection objective lens are arranged in a cassette, a luminous surface of the working electrode faces the collection objective lens, and the focus of the collection objective lens coincides with an entrance slit of the CCD grating spectrometer; the cassette is directly connected with a fixing plate of the entrance slit, and a detecting system light path is isolated from the outside; the CCD grating spectrometer and the electrochemistry equipment unit are triggered through TTL (transistor transistor logic) signals to realize synchronous operation, and the acquired electrochemical signals and ECL spectrum signals are synchronously input into the data processing and control unit.
Description
Technical field
The present invention relates to a kind of can the detection system of accurate acquisition electrogenerated chemiluminescence spectral information, belong to electroluminescent technical field of chemical detection.
Background technology
Electrogenerated chemiluminescence (Electrochemiluminescence, ECL) has electrochemistry and chemiluminescent advantage concurrently, is a kind of utilize electrochemical method excite certain material and so as to produce the analytical technology of light radiation.ECL analytical technology, without excitation source, has signal to noise ratio height, detects the sensitive and simple feature of device, has obtained extensive use in the field such as biochemical analysis, medical diagnosis.ECL radiation is generally more weak and intensity is unstable, cause that the acquisition method being difficult to adopt conventional fluorescence spectrum harvester or length scanning obtains its spectral information, therefore, ECL detecting device generally with photo-electric conversion element highly sensitive, that time response is fast and receptor area is big (such as photomultiplier tube, PMT) as detector, take the mode that ECL reaction tank is directly closely assembled in PMT front end to gather ECL efficiently and radiate overall strength, to reduce light loss, to improve signal to noise ratio and sensitivity for analysis.At present, be commercial ECL analyser or the ECL that each laboratory assembles voluntarily to detect equipment be all formed (Chem.Rev.2008 by chemiluminescent analyzer (or PMT and electric supply installation module) thereof and electrochemical apparatus simple combination, 108,2506).E1ecsys series fully automatic electric chemical illumination immunity analysis instrument that Roche Holding Ag (ROCHE) produces and the MPI Series Electrochemical luminometer that Xi'an Rui Mai Analytical Instrument Co., Ltd produces are the Typical Representatives of ECL analyser Development Status.Wherein, the clinical diagnosis of the projects such as Thyreoidine, hormone, myocardial damage, tumor markers, infectious disease, anemia and bone metabolism of being widely used by the full-automatic ECL immunity analysis instrument of E1ecsysl010 and Elecsys2010 detected based on ruthenium bipyridilium systems ECL radiation overall strength, at home and abroad obtains extensive use in big-and-middle-sized hospital.And the MPI-A type capillary electrophoresis electrochemical light-emitting detector that Xi'an Rui Mai Analytical Instrument Co., Ltd produces and MPI-E type micro-fluidic chip electrochemiluminescdetection detection instrument are the Typical Representatives of domestic goods ECL analyser.MPI-A and MPI-E type ECL detector system can the overall strength signal of simultaneous display ECL radiation and corresponding electrochemical signals, ECL basic research is helpful.
Conventional ECL detecting device all can only adopt the spectral information that the mode installing optical filter, repeatedly parallel assay between ECL reaction tank and PMT additional draws roughly ECL radiation, and measurement result repeatability is poor.This causes that conventional ECL detecting device can only carry out simple qualitative analysis research, can not accurately and in detail study spectral quality and the Changing Pattern of ECL radiation, more cannot carry out Quantitative Study based on ECL spectral information, thus greatly limit ECL to analyze the application in basic research and diagnostic field.In fact, owing to spectrally resolved form cannot be adopted to accurately distinguish the ECL radiation information in unlike signal source, the conventional ECL apparatus (including E1ecsys series fully automatic electric chemical illumination immunity analysis instrument) depending on detection ECL signal overall strength is only capable of carrying out simple one pack system analysis.
In recent years, academia has begun to attempt adopting CCD grating spectrograph or the mode that charge coupled cell (Charge-coupledDevice, CCD) and monochromator simply couple being gathered ECL spectrum (J.Am.Chem.Soc.2016,138,1947；J.Am.Chem.Soc.2015,137,11266).But, cause that incident flux is relatively low owing to ECL radiates the particularity of producing method, the reason such as the electrochemical apparatus that adopts is poor with CCD grating spectrograph synthetic operation, interference of stray light and instrument self-noise signal, the way not only signal to noise ratio that above-mentioned directly employing CCD grating spectrograph gathers ECL spectrum is low, sensitivity is poor, can not the spectrum of accurate acquisition weak ECL radiation, and repeatability is poor, it is impossible to accurately compare the spectrum change of ECL radiation under different electrochemical states or experiment condition.Therefore, the spectral type ECL research being based purely on CCD grating spectrograph and carry out also is limited in simple qualitative analysis research category.
Summary of the invention
For the deficiencies in the prior art, the present invention adopts closed optical path system of building, improves ECL radiation collecting efficiency and the method adopting TTL signal to trigger electrochemical apparatus and CCD grating spectrograph synchronous operation, construct a kind of can the detection system of accurate acquisition ECL spectral information, this system not only background noise is low, there is higher sensitivity for analysis, the spectral information of superweak ECL radiation can be detected, and can accurately compare the spectrum change of ECL radiation under different electrochemical state or experiment condition, carry out spectral type ECL quantitative analysis and multicomponent analysis research.
Technical scheme is as follows:
A kind of can the detection system of accurate acquisition ECL spectral information, radiate generator unit, spectrum detection device unit and data process and control unit including electrochemical apparatus unit, ECL；
Described ECL radiates generator unit and includes ECL pond, is provided with working electrode, reference electrode and to electrode in described ECL pond, described working electrode, reference electrode and electrode is connected with electrochemical apparatus unit respectively；
Described spectrum detection device unit includes the collection object lens for collecting ECL radiation and CCD grating spectrograph；Described CCD grating spectrograph is made up of ccd detector and grating spectrograph；The light-emitting area of described working electrode is just to described collection object lens；
Described CCD grating spectrograph and electrochemical apparatus unit process with data respectively and are connected with control unit, are connected by TTL signal line between described CCD grating spectrograph and electrochemical apparatus unit.Triggered by TTL signal between CCD grating spectrograph and electrochemical apparatus unit and realize synchronous operation, the Electrochemistry Information that the ECL spectral information gathered by CCD grating spectrograph and electrochemical apparatus unit gather is processed and control unit by synchronous transport to data, it is achieved the accurate acquisition of ECL spectral information under different electrochemical states.
According to the invention it is preferred to, described ECL radiation generator unit is additionally included in bottom and is provided with the scalable ECL pond support of loophole, and ECL pond is fixed on the support of scalable ECL pond.Described scalable ECL pond support can realize the adjustment of geometric position, ECL pond in three dimensions, it is ensured that the ECL radiation produced bottom working electrode at utmost enters CCD grating spectrograph through collecting object lens.
According to the invention it is preferred to, the bottom in described ECL pond is transparent configuration, it is further preferred that described ECL pond is all-transparent structure；
Preferably, described ECL pond is provided with electrode fixing device, described working electrode, reference electrode and electrode is fixed on electrode fixing device.
According to the invention it is preferred to, described the detection system of accurate acquisition ECL spectral information can also include magazine, described ECL radiates generator unit and collects object lens and be arranged in magazine；
Preferably, described magazine is provided with electrode cable converting interface and airway converting interface；Under ensureing the premise that light path is completely enclosed, meet electrode connect and the needs of ventilation in experimentation with extraneous.
According to the invention it is preferred to, described ECL pond is additionally provided with airway, and inside airway UNICOM ECL pond, described airway is connected to outside magazine by airway converting interface.During the detection of ECL spectral information, loading quantum dot solution in ECL pond, airway one end is positioned at below ECL pond liquid level, and the other end can connect the feeder outside magazine through airway converting interface；Arranging airway can conveniently to carrying out deoxygenation or logical oxygen in ECL pond.Preferably, described airway surface configuration has graduation mark, and breather gas outlet can be adjusted to liquid level after terminating by solution ventilation, keeps the liquid level nitrogen atmosphere such as (or oxygen certain specific gas) in experimentation.
According to the invention it is preferred to, described working electrode, reference electrode and electrode is connected with electrochemical apparatus unit respectively by electrode cable converting interface.
According to the present invention, preferably, described CCD grating spectrograph is provided with entrance slit and the fixing plate of entrance slit, and described magazine is provided with C or F interface, described collection object lens are fixed in magazine by C or F interface, and the focus collecting object lens overlaps with entrance slit.Magazine can adopt screw fixed form to be directly connected with the fixing plate of entrance slit, magazine box body is provided with C or F interface (being preferably embedded into formula C or F interface) with the fixing plate connecting portion of entrance slit and collects object lens for fixing, it is achieved ECL radiates the light path linking between generator unit and CCD grating spectrograph.Its light-emitting area is just to described collection object lens when producing ECL radiation for working electrode, and the entrance slit position of described CCD grating spectrograph is then in described gleanings mirror foci, it is ensured that ECL radiation enters CCD grating spectrograph with maximal incident luminous flux.In a word, collect object lens and supporting the use of magazine and not only whole light path can be hedged off from the outer world, be prevented effectively from the interference of veiling glare and dust, and be favorably improved the incident flux of CCD grating spectrograph, improve the sensitivity of signal to noise ratio and ECL spectral detection.
According to the present invention, preferably, described scalable ECL pond support, ECL pool surface, electrode fixing device and working electrode surface are provided with graduation mark and scalable screw thread, can accuracy controlling ECL pond and working electrode relative to the geometric position collecting object lens and entrance slit.This design not only may insure that and obtains maximal incident luminous flux by accurately regulating of geometric position, improve ECL and radiate collecting efficiency, and help to ensure that working electrode is relative to the repeatability collecting object lens and entrance slit geometric position, and then improve reliability and the repeatability of quantitative analysis results.
According to the present invention, described electrochemical apparatus unit is that the TTL signal triggering that can accept ccd detector runs, or, can directly produce TTL signal and trigger the electrochemical analyser of ccd detector synchronous operation；Can ensure that and adopt the TTL form triggered to realize electrochemical apparatus unit and the synthetic operation of spectrum detection device unit.
According to the invention it is preferred to, described ECL pond is provided with antireflection film, to reduce radiation loss.
According to the invention it is preferred to, described magazine is stainless steel, and magazine outer surface coated insulation material, magazine inner surface is fixedly installed resistant material.Magazine is possible not only to the effectively radiation of shielding external electromagnetic and the interference of veiling glare, moreover it is possible to ensure whole system normal operation under without darkroom and dust-proof experiment condition.
According to the invention it is preferred to, described ccd detector is liquid nitrogen refrigerating type ccd detector, it is possible to effectively reduce the ambient noise signal of ccd detector, improves signal to noise ratio and ECL spectral detection sensitivity.
According to the invention it is preferred to, it is computer that described data process with control unit.Data process with control unit for realizing the collection accurately of ECL radiation generator unit, spectrum detection device unit and electrochemical apparatus unit synchronous operation and spectral signal.
Beneficial effects of the present invention is as follows:
1, the present invention can accurately detect the spectral information of ECL radiation, compares the spectrum change of ECL radiation under different electrochemical states or experiment condition, carries out spectral type ECL analysis and research.
2, the whole light path system in the present invention can be hedged off from the outer world, and can gather ECL spectrum, it is not necessary to special dedusting darkroom under common lab and natural optical condition.
3, the detection system signal noise ratio in the present invention is high, the spectral information of the superweak ECL radiation of energy accurate acquisition.
4, under energy detection by quantitative different experimental conditions of the present invention, the faint change of ECL spectrum, carries out the ECL Quantitative Study of spectral type.
5, the radiation signal of energy ONLINE RECOGNITION difference ECL luminous body of the present invention, carries out multi-component ECL analysis and research.
6, the present invention can in 300-900nm or broader spectral region, in real time, dynamically detection includes the spectral information of all kinds of superweak radiation of ECL, wavelength accuracy less than 1.0nm, time of exposure be 1 millisecond controlled to 1 hour, ECL signal intensity can be carried out Accurate Determining.
Accompanying drawing explanation
Fig. 1 is that the present invention is a kind of can the agent structure schematic diagram of detection system of accurate acquisition ECL spectral information.
Wherein: 1, electrochemical analyser, 2, ECL pond, 3, CCD grating spectrograph, 4, computer, 5, magazine, 6, working electrode, 7, reference electrode, 8, to electrode, 9, scalable ECL pond support, 10, collect object lens, 11, ccd detector, 12, grating spectrograph, 13, entrance slit, 14, the fixing plate of entrance slit, 15, C or F interface, 16, TTL signal line, 17, electrode cable converting interface, 18, airway converting interface, 19, airway, 20, electrode fixing device.
Detailed description of the invention
Below by specific embodiment, the present invention will be further described, but is not limited to this.
Embodiment adopts 150 line gratings of blaze wavelength respectively 500nm and 800nm and Acton standard SP-2150 spectrogrph and PYLON-400BRX liquid nitrogen refrigerating type ccd detector composition CCD grating spectrograph.
The reason selecting 150 line gratings is in that ECL radiation belongs to molecular spectrum, and bands of a spectrum are very wide, it is not necessary to too high spectrally resolved precision.Blaze wavelength at two pieces of grating combinations of 500nm and 800nm, then can realize the spectra collection to visible region and near-infrared region ECL preferably.The reason selecting Acton standard SP-2150 spectrogrph is in that the focal length of this spectrogrph is 150 millimeters, is that same quasi-instrument mid-focal length is the shortest and spectrogrph that luminous flux is maximum, it is possible to effectively reduce light loss.Selecting the reason of PYLON-400BRX type liquid nitrogen refrigerating type ccd detector is that the detector of this model can realize the wavelength sensitive collection in 300-1000nm wide-ultra weak signal under condition of ultralow temperature.
In embodiment, electrochemical analyser used is VersaSTAT3 type electrochemical analyser.
Embodiment 1
A kind of can the detection system of accurate acquisition ECL spectral information, radiate generator unit, spectrum detection device unit and data process and control unit including electrochemical apparatus unit, ECL；
Described ECL radiates generator unit and includes ECL pond 2, is provided with working electrode 6, reference electrode 7 and to electrode 8 in described ECL pond 2, described working electrode 6, reference electrode 7 and electrode 8 is connected with electrochemical apparatus unit respectively；
Described spectrum detection device unit includes the collection object lens 10 for collecting ECL radiation and CCD grating spectrograph 3；Described CCD grating spectrograph 3 is made up of ccd detector 11 and grating spectrograph 12；The light-emitting area of described working electrode 6 is just to described collection object lens 10；
Described CCD grating spectrograph 3 and electrochemical apparatus unit process with data respectively and are connected with control unit, are linked by TTL signal line 16 between described CCD grating spectrograph 3 and electrochemical apparatus unit.
Electrochemical apparatus unit described in the present embodiment is that the TTL signal that can accept ccd detector 11 triggers the electrochemical analyser 1 run；It is computer 4 that described data process with control unit.
The bottom in the ECL pond 2 described in the present embodiment is transparent configuration.
Embodiment 2
As described in Example 1, the difference is that:
Described the detection system of accurate acquisition ECL spectral information can also including magazine 5, described ECL radiates generator unit and collects object lens 10 and be arranged in magazine 5；Described CCD grating spectrograph 3 is provided with entrance slit 13 and the fixing plate 14 of entrance slit, described magazine 5 is provided with embedded type C or F interface 15, described collection object lens 10 are fixed in magazine 5 by C or F interface 15 so that the focus collecting object lens 10 overlaps with entrance slit 13；
Described magazine 5 is provided with electrode cable converting interface 17 and airway converting interface 18；
Described ECL pond 2 is additionally provided with airway 19, and airway 19 UNICOM ECL pond 2 is internal, and it is outside that airway 19 is connected to magazine 5 by airway converting interface 18；
Described working electrode 6, reference electrode 7 and electrode 8 is connected with electrochemical apparatus unit respectively by electrode cable converting interface 17.
In the present embodiment, magazine 5 adopts screw fixed form to be directly connected with the fixing plate 14 of entrance slit, magazine 5 box body is provided with C or F interface 15 for fixing collection object lens 10 with fixing plate 14 connecting portion of entrance slit, it is achieved ECL radiates the light path linking between generator unit and CCD grating spectrograph 3.Its light-emitting area is just to described collection object lens 10 when producing ECL radiation for working electrode 6, and the entrance slit of described CCD grating spectrograph 3 13 is then in described collection object lens 10 focus, it is ensured that ECL radiation enters CCD grating spectrograph 3 with maximal incident luminous flux.
Embodiment 3
As described in Example 2, the difference is that:
Described ECL radiates generator unit and is additionally included in bottom and is provided with support 9, the ECL pond, scalable ECL pond 2 of loophole and is fixed on scalable ECL pond support 9.
Described ECL pond support 9 is referred to use the disclosed electrogenerated chemiluminescence reaction tank locus regulation device for spectra collection of Chinese patent document CN204807438U.
Embodiment 4
As described in Example 3, the difference is that:
Described ECL pond 2 is all-transparent structure；
Described scalable ECL pond support 9, surface, ECL pond 2, electrode fixing device 20 and working electrode 6 surface are provided with graduation mark and scalable screw thread, can accuracy controlling ECL pond 2 and working electrode 6 relative to the geometric position collecting object lens 10 and entrance slit 13.
Embodiment 5
As described in Example 2, the difference is that:
Described ECL pond 2 is provided with antireflection film, to reduce radiation loss；
Described magazine 5 is stainless steel, and magazine 5 outer surface coated insulation material, magazine 5 inner surface is fixedly installed resistant material.
The principle of present system:
As shown in Figure 1, solution containing ECL material and coreagent thereof is loaded in ECL pond 2, ECL radiation can be produced on working electrode 6 surface during the startup of electrochemical analyser 1, trigger CCD grating spectrograph 3 synchronous averaging by TTL signal simultaneously, ECL radiation is effectively collected in entrance CCD grating spectrograph 3 through collecting object lens 10 and entrance slit 13, the ECL spectral information gathered by CCD grating spectrograph 3 and the Electrochemistry Information gathered by electrochemical analyser 1 are then independently and simultaneously inputted in computer 4, after completing data process in computer 4, can obtain exporting the ECL spectral information under different electrochemical state, realize the on-line checking of ECL spectrum.
The grating G rotated in monochromator can change centre wavelength so that whole piece band moves, in order to obtains the ECL spectrum of different wavelength range.
Claims (10)
1. one kind can the detection system of accurate acquisition ECL spectral information, it is characterised in that this detection system includes electrochemical apparatus unit, ECL radiates generator unit, spectrum detection device unit and data and processes and control unit；
Described ECL radiates generator unit and includes ECL pond, is provided with working electrode, reference electrode and to electrode in described ECL pond, described working electrode, reference electrode and electrode is connected with electrochemical apparatus unit respectively；
Described spectrum detection device unit includes the collection object lens for collecting ECL radiation and CCD grating spectrograph；Described CCD grating spectrograph is made up of ccd detector and grating spectrograph；The light-emitting area of described working electrode is just to described collection object lens；
Described CCD grating spectrograph and electrochemical apparatus unit process with data respectively and are connected with control unit, are connected by TTL signal line between described CCD grating spectrograph and electrochemical apparatus unit.
2. according to claim 1 can the detection system of accurate acquisition ECL spectral information, it is characterised in that described ECL radiates generator unit and is additionally included in bottom and is provided with the scalable ECL pond support of loophole, and ECL pond is fixed on the support of scalable ECL pond.
3. according to claim 1 can the detection system of accurate acquisition ECL spectral information, it is characterised in that the bottom in described ECL pond is transparent configuration, it is preferred that described ECL pond is all-transparent structure；
Preferably, described ECL pond is provided with electrode fixing device, described working electrode, reference electrode and electrode is fixed on electrode fixing device.
4. according to claim 1 can the detection system of accurate acquisition ECL spectral information, it is characterised in that described the detection system of accurate acquisition ECL spectral information can also including magazine, described ECL radiates generator unit and collects object lens and be arranged in magazine；
Preferably, described magazine is provided with electrode cable converting interface and airway converting interface.
5. according to claim 4 can the detection system of accurate acquisition ECL spectral information, it is characterised in that described ECL pond is additionally provided with airway, and inside airway UNICOM ECL pond, airway is connected to outside magazine by airway converting interface.
6. according to claim 4 can the detection system of accurate acquisition ECL spectral information, it is characterized in that, described CCD grating spectrograph is provided with entrance slit and the fixing plate of entrance slit, described magazine is provided with C or F interface, described collection object lens are fixed in magazine by C or F interface, and the focus collecting object lens overlaps with entrance slit.
7. according to claim 2 can the detection system of accurate acquisition ECL spectral information, it is characterised in that described scalable ECL pond support, ECL pool surface, electrode fixing device and working electrode surface are provided with graduation mark and scalable screw thread.
8. according to claim 1 can the detection system of accurate acquisition ECL spectral information, it is characterized in that, described electrochemical apparatus unit is that the TTL signal triggering that can accept ccd detector runs, or, can directly produce TTL signal and trigger the electrochemical analyser of ccd detector synchronous operation.
9. according to claim 4 can the detection system of accurate acquisition ECL spectral information, it is characterised in that described magazine is stainless steel, and magazine outer surface coated insulation material, magazine inner surface is fixedly installed resistant material.
10. according to claim 1 can the detection system of accurate acquisition ECL spectral information, it is characterised in that it is computer that described data process with control unit.
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| CN113686933A (en) * | 2021-08-04 | 2021-11-23 | 山东大学 | Method for high-sensitively acquiring electrochemiluminescence spectrum based on light intensity type detection device |
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| CN113686933B (en) * | 2021-08-04 | 2022-04-22 | 山东大学 | Method for high-sensitively acquiring electrochemiluminescence spectrum based on light intensity type detection device |
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