CN102707072A - Human serum protein concentration testing device and method - Google Patents
Human serum protein concentration testing device and method Download PDFInfo
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- CN102707072A CN102707072A CN2012102156072A CN201210215607A CN102707072A CN 102707072 A CN102707072 A CN 102707072A CN 2012102156072 A CN2012102156072 A CN 2012102156072A CN 201210215607 A CN201210215607 A CN 201210215607A CN 102707072 A CN102707072 A CN 102707072A
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Abstract
The invention discloses a human serum protein concentration testing device and method. The device consists of a light-emitting diode array, a micro-lens array, an optical filter, a converging lens, a sample pond, an optical fiber light cone, a narrowband light filtering film, a power supply, an SSPM (Security Service Provider Module) detector, a signal processing circuit and a display module. The method comprises the following steps of: collimating light generated by the micro-lens array, filtering, converging and emitting into Zn(II) tetracarboxy phthalocyanine and a serum protein reaction solution to be detected in the sample pond; activating the reaction solution to generate fluorescent light by using a light source; acquiring and transmitting fluorescent light by using the optical fiber light cone; filtering background parasitic light by using the narrowband light filtering film; performing photoelectric conversion on a light signal by using the SSPM detector; and processing a signal output by the SSPM detector by using the signal processing circuit, and displaying the concentration value of serum proteins on a liquid display screen of the display module. The testing device disclosed by the invention has the advantages of small size, low cost, convenience for carrying and high sensitivity; and the testing method is easy to operate, and contributes to popularizing.
Description
Technical field
The invention belongs to medical detection technology, be specifically related to a kind of Human Serum Albumin's concentration determination device and method of testing.
Background technology
Protein is all Source of life, is one of material the most basic in the vital movement.Content is big in vivo, and is of a great variety, carrying the task that biosome is accomplished various biological functions, is very important and requisite for earning a bare living.In normal human's body, protein accounts for 18%, and the skin of human body, muscle, blood, internal organ main body are protein; Various hormones, enzyme, antibody all are protein in hair, bone, nail and the body; Protein is being undertaken various physiological functions, keeps the carrying out of biosome metabolic activity on the whole, is the direct expresser of biological character; Such as human serum albumins (HSA) is the abundantest transport protein of people's in-vivo content; HSA has many important physical and pharmacology function, can endogenously combine with exogenous material such as fatty acid, amino acid, hormone, zwitterion and medicine etc. with many, plays storage and transhipment effect.Human immunoglobulin(HIg) (y-G) is being played the part of important role as lymphocytic cell surface receptor in many cellular activities, in human immunity is replied, bringing into play crucial effects simultaneously.Therefore, protein is described as " building materials of human body ", " brick of life ".
Haemocyanin concentration mainly adopts haemocyanin solution measuring instrument in the measurement blood; It is widely used in blood station, ICU, operating room, gynemetrics (delivery room), surgical clinical, kidney and passes through many aspects such as section, hematology, emergency ward, ambulance, neonate's monitoring and sportsman's health check-up; Become the most important means of a lot of medicals diagnosis on disease; Be the clinical accurately and timely firsthand information that provides, Medical Instruments is absolutely necessary.
At present, have the measuring instrument of multiple haemocyanin solution concentration both at home and abroad, external Hemoglobin albuminimeter like Sweden; The main reagent strip method that adopts is measured protein concentration, and this instrument blood sampling is convenient, need not dilution, test is succinct, be very suitable for the training athlete scene tests, but it costs an arm and a leg; Cause middle and small hospital to purchase in batches; Agents useful for same bar price is also very high in addition, and is consumables, and this has increased patient's burden undoubtedly; The domestic close scientific instrument of as above Nereid company limited analyzes the 5200 type haemoglobin appearance that head factory is produced; Adopt single wavelength method to let the light of certain wavelength through the cuvette of blank solution (cyaniding high ferro standard dilution) is housed; Let the cuvette of the haemoglobin reaction solution that the light of identical wavelength generates through the hemoglobin solutions reaction of being equipped with after cyaniding high ferro and the dilution again; To analyzing through two kinds of light intensity behind cuvette that blank solution is housed and the cuvette that the haemoglobin reaction solution is housed respectively; Thereby draw the concentration of the solution of surveying, this apparatus measures time is shorter, but joins diluter 250 times of hemodilutions outside needing; And then the blood after will diluting and cyaniding high ferro are carried out hemolytic reaction as detected solution; Cause the volume of this instrument very huge, dilution and haemolysis need the long period simultaneously, make the time of the single blood sample of actual measurement increase greatly; On the other hand, compare correction owing to adopt single wavelength method need measure through the light intensity behind the cuvette that blank solution is housed, cause complicated operation, efficiency of measurement is low; In addition, twice measurement causes The ultimate results to be subject to the influence of extraneous factor (like level of skill of operator etc., extraneous flying dust etc.), directly influenced measuring accuracy.
Summary of the invention
The objective of the invention is to above-mentioned technical matters and prior art defective and deficiency, the present invention aims to provide that a kind of proving installation volume is little, cost is low, easy to carry, highly sensitive; This method of testing is simple to operate, is beneficial to the Human Serum Albumin's concentration determination device and the method for testing of popularization.
The present invention to achieve these goals, the technical solution of employing is:
A kind of Human Serum Albumin's concentration determination device is characterized in that device is made up of light emitting diode matrix, microlens array, optical filter, plus lens, sample cell, optical fiber cone, narrow-band-filter film, power supply and SSPM detector, signal processing circuit and display module, and microlens array receives the light that light emitting diode matrix sends; Optical filter is close to microlens array and is used to filter the light that passes microlens array; Plus lens links to each other with optical filter and converges the light after the filtration, and sample cell is positioned on the convergent point of plus lens, and light incides sample cell after plus lens converges; Optical fiber cone is positioned at the plus lens below; Become 90 ° of angles with the axis of plus lens, the fluorescence light wave that the optical excitation that collection and transmission sample are sent by light source produces, the narrow-band-filter film is positioned at the plus lens end; Be used for the background miscellaneous light that filter light linear light awl is gathered fluorescence; The SSPM detector links to each other with the narrow-band-filter film, converts fluorescence into voltage signal, and signal processing circuit is handled the voltage signal of SSPM detector output; Obtain the content of haemocyanin in the testing sample, the concentration value of haemocyanin is presented on the liquid crystal display of display module.
Human Serum Albumin's concentration determination method, accomplish by following step:
Step 1: get the 1mL Human Serum Albumin and mix with 3mL 1 * 10-3mol/L tetracarboxylic Phthalocyanine Zinc, haemocyanin in the sample cell and tetracarboxylic Phthalocyanine Zinc (ZnC4Pc) play chromogenic reaction;
Step 2: to reactant in the above-mentioned steps at optical alignment that light emitting diode matrix sends, filter, converge after; It is incided in the above-mentioned sample cell on the tetracarboxylic Phthalocyanine Zinc to be measured and haemocyanin reaction solution, and the tetracarboxylic Phthalocyanine Zinc to be measured in the said sample cell of light source activation is the fluorescence of 475nm with the reaction solution of haemocyanin generation wavelength;
Step 3: the fluorescence that produces in the optical fiber cone acquisition step two, make it pass through its background miscellaneous light of narrow-band-filter film filtering, and the fluorescence after will filtering is transferred to the SSPM detector, convert voltage signal into;
Step 4: adopt signal processing circuit that the signal of SSPM detector output is handled; Through comparing the corresponding voltage amplitude value of variable concentrations haemocyanin standard model; Formulate typical curve; According to the pairing voltage amplitude value of testing sample, obtain the content of haemocyanin in the testing sample, and the concentration value of haemocyanin is shown on the liquid crystal display of display module.
Wherein, said tetracarboxylic Phthalocyanine Zinc solution is by 4-carboxyl phthalic anhydride, zinc chloride, ammonium molybdate and urea synthetic, and Phthalocyanine Zinc has tangible catalytic activity in analytical approachs such as fluorescence, chemiluminescence.The present invention just is being based on the resonance Rayleigh scattering principle, and Phthalocyanine Zinc is combined with human body protein, makes the resonance Rayleigh intensity of specific wavelength sharply strengthen, thereby accomplishes the quantitative measurement to protein.
Said light emitting diode matrix is to constitute by adopting Micrometer-Nanometer Processing Technology integrated a large amount of light emitting diodes on semi-conductor chip to arrange.The main wired arrangement mode of the array way of light emitting diode, face arrangement mode and three-dimensional arrangement mode.Adopt the face arrangement mode among the present invention; The characteristics of face arrangement mode are: illuminating area is little, light output is concentrated, illuminance is high, be applicable to that plane of illumination is less and relatively concentrate but the brightness value requirement than higher irradiation place, therefore; Adopt light emitting diode matrix among the present invention; Can reduce the volume of light source, reduce cost, and raising excites light intensity.
In apparatus of the present invention, the light that light emitting diode matrix sends is collimated into directional light through microlens array, again through the processing that filters of rear end optical filter.Said microlens array (MLA) is a series of apertures at the microminiature lens of several microns to hundreds of micron by the array that necessarily rearranges; It is the important and basic optical element that constitutes micro-optical systems; Have size little, be convenient to extensive manufacturing, loss little, can be made into array format, advantage such as specific function arranged, be widely used in the sub-wavelength grate structure of micro-optical systems, optical parallel disposal system, wide field and infrared imaging system, optically filtering and material processing system and antireflection and polarization state control etc.Said micro lens array can be refractive micro lens array or diffraction type micro lens array.Adopt the diffraction type microlens array among the present invention.
Said optical fiber cone is to be processed through regularly arranged, heating, the series of process such as merging, reverse, draw awl of pressurizeing by up to ten thousand optical fibers; Wherein each root fiber is made up of the core glass of high index of refraction and the foreskin glass of low-refraction, and incident light passes to the other end according to the end of total reflection principle from every fiber.The Optical Fiber Transmission element comprises fibre faceplate, optical fiber image inverter, optical fiber cone.Optical fiber cone is a kind of special shape of fibre faceplate.Fibre faceplate be by tens million of diameters be 5～6 microns light transmitting fiber regularly arranged after, the fusion of heating, pressurize forms.It optically has zero thickness, and very high light collecting light ability and resolution are arranged, and can transmit high-definition image undistortedly, is the photoelectronic imaging and the image transmission apparatus of superior performance.Therefore, we adopt, and widely used optical fiber cone transmits light collection on the market, and its receiving angle can reach 180 °, has improved receiving efficiency.
Said solid-state photomultiplier (SSPM) is a kind of novel high speed photoelectric device.Solid-state photomultiplier adopts the face battle array to arrange, and the avalanche photo diode (APD) of Geiger mode of operation is made.Each APD is equivalent to a minicell, adopts unique trench technique that these APD batteries are carried out isolation, and the electric current output of all APD batteries is together in parallel, the response signal of all APD batteries that the output signal is integrated.Each APD battery is all with Geiger pattern work, so each unit can be as the photon detector of a numeral, and has very high internal gain and fast response characteristic.This architecture design not only has functional characteristics such as traditional photomultiplier is highly sensitive, speed is fast, also has following characteristics simultaneously: operating on low voltage; Do not need high-voltage power supply, high-gain, insensitive to magnetic field; Solid state device, structure are tightly short, and size is little; Be fit to multiple environment and use, be very easy to be coupled with scintillator, optical fiber etc.
Said signal processing circuit is made up of signal amplification circuit, AD converter, single-chip microcomputer, display module, keyboard load module, data storage, RS232 etc.The signal of SSPM output carries out the AD conversion after amplifying circuit amplifies, by the single-chip microcomputer calculation process, concentration value is presented on the LCDs of display module.
The substantive distinguishing features that the present invention gives prominence to significant beneficial effect is: with prior art relatively, proving installation volume of the present invention is little, cost is low, easy to carry, highly sensitive; Method of testing of the present invention is simple to operate, is beneficial to and applies.
Description of drawings
Fig. 1 is the structure diagram of a kind of Human Serum Albumin's concentration determination of the present invention device.
Fig. 2 is tetracarboxylic Phthalocyanine Zinc and the emission spectrum of Human Serum Albumin's reaction solution under the 405nm excitation light irradiation.
Fig. 3 is the signal processing circuit theory diagram.
Fig. 4 is haemocyanin concentration and its corresponding voltage Value Data that experiment obtains.
Fig. 5 is protein concentration-voltage standard curve.
Embodiment
Below in conjunction with Fig. 1～Fig. 4 the present invention is elaborated:
As shown in Figure 1: a kind of Human Serum Albumin's concentration determination device, form by light emitting diode matrix 1, microlens array 2, optical filter 3, plus lens 4, sample cell 5, optical fiber cone 6, narrow-band-filter film 7, power supply 8 and SSPM detector 9, signal processing circuit 10 and display module 11.
The light that light emitting diode matrix 1 sends passes next-door neighbour's microlens array 2 backs by collimation, and optical filter 3 is close to microlens array 2 and is used to filter the light that passes microlens array 2, and plus lens 4 links to each other with optical filter 3; Converge the light after the filtration, the tetracarboxylic Phthalocyanine Zinc is housed and haemocyanin reaction solution sample cell 5 is positioned on the convergent point of plus lens 4, light incides sample cell 5 after plus lens 4 converges; The optical excitation that sample in the sample cell 5 is sent by light source produces fluorescence; Its emission spectrum is as shown in Figure 2, and optical fiber cone 6 is positioned at plus lens 4 belows, becomes 90 ° of angles with the axis of plus lens 4; Gather and transmit the fluorescence of above-mentioned sample generation; Narrow-band-filter film 7 is positioned at plus lens 4 ends, is used for the background miscellaneous light that filter light linear light awl 6 is gathered fluorescence, and SSPM detector 9 links to each other with narrow-band-filter film 7; Convert fluorescence into voltage signal; Signal processing circuit is handled the voltage signal of SSPM detector 9 outputs, obtains the content of haemocyanin in the testing sample, and the concentration value of haemocyanin is presented on the liquid crystal display of display module.
Use the method for testing of Human Serum Albumin's concentration determination device, accomplish by following step:
Step 1: get the 1mL Human Serum Albumin and mix, make haemocyanin and tetracarboxylic Phthalocyanine Zinc in the sample cell play chromogenic reaction with 3mL 1 * 10-3mol/L tetracarboxylic Phthalocyanine Zinc;
Step 2: to reactant in the above-mentioned steps at optical alignment that light emitting diode matrix sends, filter, converge after; It is incided in the above-mentioned sample cell on the tetracarboxylic Phthalocyanine Zinc to be measured and haemocyanin reaction solution, and the tetracarboxylic Phthalocyanine Zinc to be measured in the said sample cell of light source activation is the fluorescence of 475nm with the reaction solution of haemocyanin generation wavelength;
Step 3: the fluorescence that produces in the optical fiber cone acquisition step two, make it pass through its background miscellaneous light of narrow-band-filter film filtering, and the fluorescence after will filtering is transferred to the SSPM detector, convert voltage signal into;
Step 4: adopt signal processing circuit that the signal of SSPM detector output is handled; Through comparing the corresponding voltage amplitude value of variable concentrations haemocyanin standard model; Formulate typical curve; According to the pairing voltage amplitude value of testing sample, obtain the content of haemocyanin in the testing sample, and the concentration value of haemocyanin is shown on the liquid crystal display of display module.
Concrete test sample is following:
Adopt the measurement that experimentizes of the haemocyanin solution of tetracarboxylic Phthalocyanine Zinc solution and 50mg/mL, 40mg/mL, 30mg/mL, 25mg/mL, 20mg/mL, 10mg/mL, 5mg/mL, the 1mg/mL of 2 * 10-4mol/L; Each concentration duplicate measurements 8 times, the data that obtain are as shown in table 1.
Table 12.0 * 10-4mol/L tetracarboxylic Phthalocyanine Zinc solution and haemocyanin experimental data
Concentration and corresponding voltage data thereof by the experiment of the tetracarboxylic Phthalocyanine Zinc solution of 2.0 * 10-4mol/L obtains are seen Fig. 4.Visible by Fig. 4, when haemocyanin solution concentration during greater than 10g/L, curve better linear.And serum albumin levels is between 40-50mg/ml in the human body, therefore fitting formula in concentration 10-50mg/ml, and linear relationship is good.
Carry out data processing with the least square method his-and-hers watches; The mensuration of experimental data receives the influence of various empirical factors, and for improving the accuracy of match linear relation, the mean value of respectively organizing data is adopted in match; Obtaining linear relation at last is: y=0.711x+60.54, its correlation coefficient r=0.9939.Above-mentioned linear relationship as protein concentration-voltage standard curve, is seen Fig. 5.
Claims (5)
1. Human Serum Albumin's concentration determination device is characterized in that device is made up of light emitting diode matrix, microlens array, optical filter, plus lens, sample cell, optical fiber cone, narrow-band-filter film, power supply and SSPM detector, signal processing circuit and display module; Said microlens array receives the light that light emitting diode matrix sends, and optical filter is close to microlens array and is used to filter the light that passes microlens array, and plus lens links to each other with optical filter and converges the light after the filtration; Sample cell is positioned on the convergent point of plus lens; Light incides sample cell after plus lens converges, optical fiber cone is positioned at the plus lens below, becomes 90 ° of angles with the axis of plus lens; The fluorescence light wave that the optical excitation that collection and transmission sample are sent by light source produces; The narrow-band-filter film is positioned at the plus lens end, is used for the background miscellaneous light that filter light linear light awl is gathered fluorescence, and the SSPM detector links to each other with the narrow-band-filter film; Convert fluorescence into voltage signal; Signal processing circuit is handled the voltage signal of SSPM detector output, obtains the content of haemocyanin in the testing sample, and the concentration value of haemocyanin is presented on the liquid crystal display of display module.
2. according to claim 1 described Human Serum Albumin's concentration determination device, the arrangement mode that it is characterized in that said light emitting diode matrix is line arrangement mode, face arrangement mode or three-dimensional arrangement mode.
3. Human Serum Albumin's concentration determination device according to claim 1 is characterized in that said light emitting diode matrix sends the light that wavelength is 405nm.
4. Human Serum Albumin's concentration determination device according to claim 1, the centre wavelength that it is characterized in that said narrow-band-filter film is 475nm.
5. the method for testing of claim 1 described Human Serum Albumin's concentration determination device is characterized in that being accomplished by following step:
Step 1: get the 1mL Human Serum Albumin and mix, make haemocyanin and tetracarboxylic Phthalocyanine Zinc in the sample cell play chromogenic reaction with 3mL 1 * 10-3mol/L tetracarboxylic Phthalocyanine Zinc;
Step 2: to reactant in the above-mentioned steps at optical alignment that light emitting diode matrix sends, filter, converge after; It is incided in the above-mentioned sample cell on the tetracarboxylic Phthalocyanine Zinc to be measured and haemocyanin reaction solution, and the tetracarboxylic Phthalocyanine Zinc to be measured in the said sample cell of light source activation is the fluorescence of 475nm with the reaction solution of haemocyanin generation wavelength;
Step 3: the fluorescence that produces in the optical fiber cone acquisition step two, make it pass through its background miscellaneous light of narrow-band-filter film filtering, and the fluorescence after will filtering is transferred to the SSPM detector, convert voltage signal into;
Step 4: adopt signal processing circuit that the signal of SSPM detector output is handled; Through comparing the corresponding voltage amplitude value of variable concentrations haemocyanin standard model; Formulate typical curve; According to the pairing voltage amplitude value of testing sample, obtain the content of haemocyanin in the testing sample, and the concentration value of haemocyanin is shown on the liquid crystal display of display module.
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| CN106092902A (en) * | 2016-06-30 | 2016-11-09 | 上海市奉贤区中心医院 | Medical scanning devices |
| CN108489936A (en) * | 2018-04-08 | 2018-09-04 | 吉林省富生医疗器械有限公司 | A kind of domestic type cystatin C concentration Quick testing instrument and its assay method |
| CN108918910A (en) * | 2018-08-02 | 2018-11-30 | 中南大学 | A method of monitoring two-dimensional material suspension or gel rate travel |
| GB2568307A (en) * | 2017-11-14 | 2019-05-15 | Stratec Biomedical Ag | Spectral excitation device |
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106092902A (en) * | 2016-06-30 | 2016-11-09 | 上海市奉贤区中心医院 | Medical scanning devices |
| GB2568307A (en) * | 2017-11-14 | 2019-05-15 | Stratec Biomedical Ag | Spectral excitation device |
| CN108489936A (en) * | 2018-04-08 | 2018-09-04 | 吉林省富生医疗器械有限公司 | A kind of domestic type cystatin C concentration Quick testing instrument and its assay method |
| CN108489936B (en) * | 2018-04-08 | 2024-03-19 | 吉林省富生医疗器械有限公司 | Household cystatin C concentration rapid tester and testing method thereof |
| CN108918910A (en) * | 2018-08-02 | 2018-11-30 | 中南大学 | A method of monitoring two-dimensional material suspension or gel rate travel |
| CN108918910B (en) * | 2018-08-02 | 2020-07-28 | 中南大学 | Method for monitoring two-dimensional material suspension or gel moving speed |
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