CN103472034B - Blood cell analysis chip, analyzer and analysis method - Google Patents
Blood cell analysis chip, analyzer and analysis method Download PDFInfo
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Abstract
The invention belongs to the technical field of blood cell analysis, and particularly relates to a blood cell analysis chip, an analyzer and an analysis method. The blood cell analysis chip of the present invention comprises: the leucocyte analysis chip comprises a peroxidase detection chip and a basophil detection chip, the peroxidase detection chip utilizes laser scattering and peroxidase staining technologies to classify cells, the basophil detection chip adopts a biological dissolution and electrical impedance method to detect basophil, and the erythrocyte, platelet or hemoglobin analysis chip adopts a dual-angle laser method to detect erythrocytes and platelets and an erythrocyte optical scattering method to detect hemoglobin. The blood cell analysis chip, the blood cell analyzer and the blood cell analysis method have the advantages of simple structure, small volume, low cost, convenience in operation, easiness in maintenance and transportation, disposable chip and the like. < pb pnum = "1"/>)
Description
Technical field
The invention belongs to blood cell analysis technical field, particularly relate to a kind of blood cell analysis chip, analysis and analytical approach.
Background technology
The information that blood cell analysis obtains can contribute to the disease diagnosed, antidiastole is relevant with hematological system, contribute to analysing patient's condition, observe the curative effect, judging prognosis, for prevent disease provides foundation, instruct clinical application and carry out clinic study, therefore haemocyte inspection (i.e. routine blood test) becomes first of three large routine inspections in clinical examination (routine blood test, routine urinalysis, just routine), and its clinical practice is also extensive.After nineteen fifty-three Mr. Ku Erte invention electrical impedance method blood-counter system, various Automatic blood cell analyzer is come out one after another, and blood cell analysis technology is developed rapidly.
At present for blood cell analysis, generally carried out under given conditions through specific people by commercially available cellanalyzer.Existing cellanalyzer is bulky, expensive, complicated operation, need special messenger to use and carry out periodic maintenance, the measurement reagent price supporting with it is also more expensive, generally be applicable to the more hospital inspection section office comparatively concentrated of test samples, for different medical units such as village health posts, often sample size is little, and sample disperses very much on time dimension, there is the situation of significant discomfort, therefore conventional inspection kind equipment shortcoming should be overcome, meet the demand of different medical unit to measuring means again, current urgent need develops portability, simple operation, the cellanalyzer of just-in-time of reporting the result, be applicable to Site Detection, emergency analysis, domestic. applications and primary care.
Summary of the invention
The invention provides a kind of blood cell analysis chip, analyser and analytical approach, be intended to solve that existing cellanalyzer is bulky, expensive, complicated operation, need special messenger to use and carry out the technical matters of periodic maintenance.
Technical scheme provided by the invention is: a kind of blood cell analysis chip, comprise: leukocyte analysis chip and red blood cell, blood platelet or hemoglobin analysis chip, described leukocyte analysis chip comprises peroxidase detection chip and basophilic granulocyte detection chip, described peroxidase detection chip utilizes laser light scattering and peroxidase staining technique to carry out cell classification, described basophilic granulocyte detection chip adopts biological dissolution and electrical impedance method to detect basophilic granulocyte, described red blood cell, blood platelet or hemoglobin analysis chip adopt two multi-angle laser method to detect red blood cell and blood platelet, and adopt red blood cell optical scattering method to detect haemoglobin.
Technical scheme of the present invention also comprises: described peroxidase detection chip is provided with liquid storage tank, waste liquid pool, detection zone and dosing assemblies, described liquid storage tank is for storing detection reagent, described waste liquid pool is for storing the blood sample through detecting, described detection zone carries out laser beam irradiation for making the leucocyte of blood sample through detection zone, classified by light scattering and the different blood sample to detecting of cell size, described dosing assemblies is used for making corresponding fluids in each liquid storage tank quantitative.
Technical scheme of the present invention also comprises: described basophilic granulocyte detection chip is provided with liquid storage tank, waste liquid pool, detection zone and dosing assemblies, described liquid storage tank is used for storage of blood sample and dilution, described waste liquid pool is for storing the blood sample through detecting, the cell that described detection zone is used for blood sample adopts electrical impedance technology to detect according to cell size through detection zone, calculated the Effective Numerical of leucocyte bare nucleus and basophilic granulocyte by electronic impulse size and number, described dosing assemblies is used for making corresponding fluids in each liquid storage tank quantitative.
Technical scheme of the present invention also comprises: described red blood cell, blood platelet or haemoglobin detection chip are provided with liquid storage tank, waste liquid pool, detection zone and dosing assemblies, described liquid storage tank is used for storage of blood sample and dilution, described waste liquid pool is for storing the blood sample through detecting, described detection zone carries out laser beam irradiation for making the leucocyte of blood sample through detection zone, classified by light scattering and the different blood sample to detecting of cell size, described dosing assemblies is used for making corresponding fluids in each liquid storage tank quantitative.
Technical scheme of the present invention also comprises: described leukocyte analysis chip and red blood cell, blood platelet or hemoglobin analysis chip adopt material to comprise quartz, glass, monocrystalline silicon or macromolecule polymeric material; Described liquid storage tank sample introduction adopts Micropump, electrokinetic injection, positive pressure to drive sample introduction, Ngatively pressurized sampling or electric osmose sample introduction various ways.
Another technical scheme provided by the invention is: a kind of cellanalyzer, comprise blood cell analysis chip as claimed in claim 1, detecting unit and signal processing system, described blood cell analysis chip, detecting unit is connected successively with signal processing system, described blood cell analysis chip is for carrying haemocyte, described detecting unit is used for applying laser beam to detection zone and irradiates haemocyte, described signal processing system applies steady current to upper and lower two ends, detection zone, detect each cell and produce the electronic impulse proportional with cell volume, according to the threshold value of setting, distinguish leucocyte bare nucleus and basophilic granulocyte, provide total white blood cells and basophilic granulocyte Effective Numerical accurately, or described signal processing system adopts high angle scattered light to measure cell peroxidase content situation, endoerythrocytic hemoglobin concentration, content of hemoglobin mean value or measurement cell refractive index, described signal processing system adopts low angle scattered light to measure cell volume or size.
The another technical scheme that the present invention takes is: a kind of blood cell analysis method, comprising:
Step a: utilize laser light scattering and peroxidase staining technique to classify to cell;
Step b: adopt biological dissolution technology and electrical impedance method to detect basophilic granulocyte;
Step c: adopt two multi-angle laser method to detect red blood cell and blood platelet, and adopt red blood cell optical scattering method to detect haemoglobin.
Technical scheme of the present invention also comprises: described step a comprises: sample introduction is to liquid storage tank S1, R1, R2 respectively for anticoagulation, dilution, clean-out system and formaldehyde isotonic liquid, and whole blood carries out suitably dilution and the short time hatches; Mix and heat with hydrogen peroxide and four chloro-naphthols through liquid storage tank R5, R6 respectively, intracellular peroxidation thing enzyme decomposition of hydrogen peroxide produces oxonium ion, and oxonium ion makes four chloro-naphthols develop the color and precipitate and is positioned enzyme reaction position; Sheath fluid sample introduction is to liquid storage tank R7, and through the blood sample of chemical staining under the effect of sheath fluid stream, leucocyte is in line and successively by detection zone A1; Flow into waste liquid pool W1 through laser beam irradiation, classify according to light scattering and cell size difference.
Technical scheme of the present invention also comprises: described step b comprises: anticoagulation sample introduction is to liquid storage tank S2, special basophilic granulocyte dilution sample introduction is to liquid storage tank R3, quantitative anticoagulation and quantitative dilution flow out from respective liquid storage tank and mix, sheath fluid sample introduction is to liquid storage tank R8, treated blood sample is under the effect of quantitative sheath fluid stream, leucocyte is in line and successively by detection zone A2, A2 upper and lower two ends in detection zone apply steady current, detect through electrical impedance and flow into waste liquid pool W2, according to threshold value setting through signal processing system Treatment Analysis, distinguish leucocyte bare nucleus and basophilic granulocyte, provide total white blood cells and basophilic granulocyte Effective Numerical accurately.
Technical scheme of the present invention also comprises: in described step c, described pair of multi-angle laser method detects to comprise and uses low angle scattered light and high angle scattered light, wherein, described low angle scattered light is for measuring erythrocyte volume, high angle scattered light measures each endoerythrocytic hemoglobin concentration and content of hemoglobin mean value, on platelet count is analyzed, high angle scattering optical power measurement cell refractive index, low angle scattering optical power measurement cell size.
Technical scheme tool of the present invention has the following advantages or beneficial effect: the blood cell analysis chip of the embodiment of the present invention, analyser and analytical approach carry out somatotype to haemocyte in anticoagulation, have that structure is simple, volume is little, cost is low, easy to operate, easy care, easily transport, chip mistake and the advantage such as discardable, meet the demand for development of analytical instrument microminiaturization, integrated and portability, be applicable to the uses such as hospital, clinic, community and individual family.
Accompanying drawing explanation
Accompanying drawing 1 is the structural representation of the blood cell analysis chip of the embodiment of the present invention;
Accompanying drawing 2 is structural representations of the peroxidase detection chip of the blood cell analysis chip of the embodiment of the present invention;
Accompanying drawing 3 is structural representations of the basophilic granulocyte detection chip of the blood cell analysis chip of the embodiment of the present invention;
Accompanying drawing 4 is structural representations of the red blood cell of the blood cell analysis chip of the embodiment of the present invention, blood platelet or haemoglobin detection chip;
Accompanying drawing 5 is structural representations of the cellanalyzer of the embodiment of the present invention;
Accompanying drawing 6 is embody rule structural representations of the cellanalyzer of the embodiment of the present invention;
Accompanying drawing 7 is process flow diagrams of the blood cell analysis method of the embodiment of the present invention.
Embodiment
In order to make object of the present invention, technical scheme and advantage clearly understand, below in conjunction with drawings and Examples, the present invention is further elaborated.Should be appreciated that specific embodiment described herein only in order to explain the present invention, be not intended to limit the present invention.
Referring to Fig. 1, is the structural representation of the blood cell analysis chip of the embodiment of the present invention.The blood cell analysis chip of the embodiment of the present invention comprises leukocyte analysis chip and red blood cell, blood platelet or hemoglobin analysis chip.
Wherein, leukocyte analysis chip comprises: peroxidase detection chip and basophilic granulocyte detection chip.Refer to Fig. 2, Fig. 2 is the structural representation of the peroxidase detection chip of the blood cell analysis chip of the embodiment of the present invention.Peroxidase detection chip is provided with liquid storage tank, waste liquid pool, detection zone and dosing assemblies.Liquid storage tank is for storing detection reagent, waste liquid pool is for storing the blood sample through detecting, detection zone carries out laser beam irradiation for making the leucocyte of blood sample through detection zone, different thus classify to the blood sample detected by light scattering and cell size, dosing assemblies is used for making corresponding fluids in each liquid storage tank quantitative.In embodiments of the present invention, liquid storage tank arranges at least one, its can be used for storing cell cleaned and destroys detection reagent, make cell carry out the zymolytic detection reagent of superoxide and sheath fluid.Peroxidase detection chip utilizes laser light scattering and peroxidase staining technique to carry out cell classification, peroxidase detection chip carries out cell classification detailed process: anticoagulation, dilution, clean-out system and formaldehyde isotonic liquid difference sample introduction are to liquid storage tank S1, R1, R2, wherein, isotonic solution is in the various solution used in clinical or Physiological Experiment, what its osmotic pressure was equal with plasma osmolarity is called isotonic solution, whole blood carries out suitably dilution and the short time hatches, now cell is destroyed by clean-out system, leucocyte slurry endoenzyme is fixed, by liquid storage tank R5, R6 mixes with hydrogen peroxide and four chloro-naphthols and heats respectively, now intracellular peroxidation thing enzyme decomposition of hydrogen peroxide produces oxonium ion, oxonium ion makes four chloro-naphthols develop the color and precipitate and is positioned enzyme reaction position, sheath fluid sample introduction is to R7 liquid storage tank, through the blood sample of chemical staining under the effect of sheath fluid stream, leucocyte is in line and flows into waste liquid pool W1 by detection zone A1 through laser beam irradiation one by one, be classified due to light scattering and cell size difference, dosing assemblies C1 to C7 is used for making corresponding fluids in each liquid storage tank quantitative.Appears as negative reaction without black-and-blue particle in cell after dyeing, occur that the black particle of fine particle or sparse distribution is weak positive reaction, occur that the thick and intensive particle of black is strong positive reaction.All kinds of leucocyte is to the reaction of peroxidase: early stage myeloblast is negative, each stage that early young grain is later all contains peroxidase, and strengthen gradually along with its content of maturation of cell, eosinophil has the strongest peroxidase, neutrophil leucocyte contains stronger peroxidase, and basophilic granulocyte is not containing this enzyme; In monocyte system, except the early stage primitive stage, inmature monokaryon and monocyte there will be more weak peroxidase reaction; Lymphocyte, erythroneocytosis, megacaryocyte etc. are all peroxidase negative reaction.To sum up, in blood five kinds of leukocytic peroxidase activities put in order into: eosinophil > neutrophil leucocyte > monocyte, lymphocyte and basophilic granulocyte are all without peroxidase.Due to after enzyme reaction in intensity difference (feminine gender, the weak positive, strong positive) and cell volume difference in size, when cell passes through detection zone A1 through laser beam irradiation, produce the scattered light of varying strength, each cell produces histochemical staining result and light scattering result two signals.
At the scatter diagram that peroxidase detection chip produces, transverse axis represents the cell peroxidase content situation obtained 7 ° of laser light scattering absorptances, and peroxidase strong positive cell is positioned at right-hand member; The longitudinal axis is the cell volume size that low angle scattered light 2.8 ° is measured, and the expression scattered light signal being positioned at top is strong, and cell volume is large.Except basophilic granulocyte, the cell of each type is recorded in specific position according to their feature, eosinophil, neutrophil leucocyte, monocyte and lymphocyte can be distinguished in setting range, and calculate total white blood cells and classification in conjunction with basophilic granulocyte detection chip result.To sum up, cell is divided into five classes by peroxidase detection chip: the 1) eosinophil of the most strong positive of peroxidase; 2) neutrophilic segmented granulocyte of peroxidase strong positive; 3) comparatively large, the p+ monocyte of peroxidase of volume; 4) lymphocyte of small volume, peroxidase feminine gender; 5) volume is greater than lymphocyte and the maxicell that is unstained of peroxidase feminine gender, and this type of cell increases prompting naivety or original various types of cells may occur.
Referring to Fig. 3, is the structural representation of the basophilic granulocyte detection chip of the blood cell analysis chip of the embodiment of the present invention.The basophilic granulocyte detection chip of the embodiment of the present invention is provided with liquid storage tank, waste liquid pool, detection zone and dosing assemblies.Liquid storage tank is used for storage of blood sample and dilution, and waste liquid pool is for storing the blood sample through detecting, and aperture is for calculating the Effective Numerical of leucocyte bare nucleus and basophilic granulocyte.Wherein, liquid storage tank quantity is at least one, is respectively used to store anticoagulation, special basophilic granulocyte dilution and sheath fluid, and in embodiments of the present invention, basophilic granulocyte detection chip used diluent is available reagent.The detailed process that basophilic granulocyte detection chip carries out detecting is: anticoagulation sample introduction is to S2 liquid storage tank, special basophilic granulocyte dilution sample introduction is to R3 liquid storage tank, quantitative anticoagulation and quantitative dilution flow out from respective liquid storage tank and realize mixing, special basophilic granulocyte reagent is by the leucocyte removing cell membrane except basophilic granulocyte, make its bare nucleus and smaller volume, and erythrocytolysis, only basophilic granulocyte keeps original state, volume is obviously greater than the leucocyte of other types, sheath fluid sample introduction is to R8 liquid storage tank, treated blood sample is under the effect of quantitative sheath fluid stream, leucocyte is in line and one by one by detection zone A2, A2 upper and lower two ends in detection zone apply steady current, detect (namely each cell produces the electronic impulse proportional with cell volume) through electrical impedance and flow into waste liquid pool W2, according to threshold value setting through signal processing system Treatment Analysis, distinguish leucocyte bare nucleus and basophilic granulocyte, provide total white blood cells and basophilic granulocyte Effective Numerical more accurately.In embodiments of the present invention, basophilic granulocyte detection chip adopts biological dissolution technology and electrical impedance method to detect basophilic granulocyte, further raising detection perform, can also adopt optical detection to detect, and dosing assemblies C8 to C11 is used for making corresponding fluids in each liquid storage tank quantitative.
Referring to Fig. 4, is the structural representation of the red blood cell of the blood cell analysis chip of the embodiment of the present invention, blood platelet or haemoglobin detection chip.Red blood cell, blood platelet or haemoglobin detection chip are provided with liquid storage tank, waste liquid pool, detection zone and dosing assemblies.Liquid storage tank is used for storage of blood sample and dilution, and waste liquid pool is for storing the blood sample through detecting, and detection zone is used for making red blood cell by carrying out laser irradiation during detection zone.Wherein, liquid storage tank quantity is at least one, is respectively used to store anti-freezing blood sample, dilution and sheath fluid.Red blood cell, blood platelet or haemoglobin detection chip detect red blood cell and adopt two multi-angle laser method, microcyte, bib and blood platelet are more easily differentiated, greatly improve accuracy and the precision of blood platelet mensuration, and adopt the direct Measuring hemoglobin of red blood cell scattered light.The detailed process that red blood cell and the red analysis chip of blood platelet carry out detecting is: anticoagulation sample introduction is to liquid storage tank S3, special dilution sample introduction is to liquid storage tank R4, quantitative anticoagulation and quantitative dilution flow out from respective liquid storage tank and realize mixing, sheath fluid sample introduction is to liquid storage tank R9, treated blood sample is under the effect of quantitative sheath fluid stream, red blood cell is in line and flows into waste liquid pool W3 by detection zone A3 one by one, laser irradiates each red blood cell scanning through detection zone A3, and low angle scattered light 2.8 ° is for measuring cell volume; High angle scattered light 7 ° measures each endoerythrocytic hemoglobin concentration (MCH) and content of hemoglobin mean value MCHC, and calculates the haemoglobin dispersion of distribution (HDW) value in red blood cell.Platelet count analysis have employed two-dimentional optical triangulation method too, high angle scattered light 7 ° can measure cell refractive index (RI), low angle scattered light 2.8 ° can measure cell size, two-dimensional scattering figure draws platelet counts and correlation parameter, blood platelet of all sizes in 1-60fl can be measured, also can measure blood platelet content content.Dosing assemblies C12 to C15 is used for making corresponding fluids in each liquid storage tank quantitative.
In the blood cell analysis chip embodiment of invention, sample introduction can adopt Micropump, electrokinetic injection, positive pressure to drive the various ways such as sample introduction, Ngatively pressurized sampling, electric osmose sample introduction, and chip can by quartz, glass, monocrystalline silicon, macromolecule polymeric material as making such as polymetylmethacrylate, polydimethylsiloxane, polycarbonates; In addition, in order to alleviate or avoid chip microchannel surface to Hemadsorption, chip microchannel inside surface can through ad hoc fashion modification, or chip material modification, or adds suitable adjuvant in solution.
Referring to Fig. 5, is the structural representation of the cellanalyzer of the embodiment of the present invention.The cellanalyzer of the embodiment of the present invention comprises blood cell analysis chip, detecting unit and signal processing system.Blood cell analysis chip, detecting unit are connected successively with signal processing system, blood cell analysis chip is for carrying haemocyte, detecting unit is used for applying laser beam to detection zone and irradiates haemocyte, signal processing system applies steady current to the upper and lower two ends of aperture, detect each cell and produce the electronic impulse proportional with cell volume, according to the threshold value of setting, distinguish leucocyte bare nucleus and basophilic granulocyte, provide total white blood cells and basophilic granulocyte Effective Numerical more accurately; Or described signal processing system adopts high angle scattered light measure cell peroxidase content situation, endoerythrocytic hemoglobin concentration, content of hemoglobin mean value or measure cell refractive index, described signal processing system adopts low angle scattered light to measure cell volume or size.
Referring to Fig. 6, is the embody rule structural representation of the cellanalyzer of the embodiment of the present invention.The cellanalyzer of the embodiment of the present invention comprises collimation lens 2, condenser lens 3, spectroscope 4, diaphragm 5, photodetector 6 and blood cell analysis chip 7.Blood cell analysis chip 7 is blood cell analysis chip described in Fig. 1, for carrying haemocyte 8, light source 1 is irradiated to the haemocyte 8 of carrying on blood cell analysis chip 7 successively by collimation lens 2, condenser lens 3, then by collimation lens 2 and spectroscope 4, the other two ends of collimation lens 2 and spectroscope 4 are all connected with diaphragm 5 and photodetector 6.The cellanalyzer of the embodiment of the present invention is except being applied to medical industry, every relate to requirement measurement of species particle diameter and liquid in atomic quantity carry out quantitatively and qualitatively analyzing, can measure with this instrument, namely this instrument can develop into laser particle size analyzer, such as, in physico-chemical analysis in pure water, measure the content of its impurity and bacterium; The measurement of the degree of purity of various industrial high purity liquid, demarcate, to microelectronic component, the production of integrated circuit, quality control, pharmaceutical industry and chemical industry, Food Hygiene Surveillance etc. have realistic meaning.
Referring to Fig. 7, is the process flow diagram of the blood cell analysis method of the embodiment of the present invention.The blood cell analysis method of the embodiment of the present invention comprises:
Step 100: utilize laser light scattering and peroxidase staining technique to classify to cell;
In step 100, utilize laser light scattering and peroxidase staining technique to carry out classification to cell to be specially: anticoagulation, dilution, clean-out system and formaldehyde isotonic liquid difference sample introduction are to liquid storage tank S1, R1, R2, whole blood carries out suitably dilution and the short time hatches, now cell is destroyed by clean-out system, leucocyte slurry endoenzyme is fixed, by liquid storage tank R5, R6 mixes with hydrogen peroxide and four chloro-naphthols and heats respectively, now intracellular peroxidation thing enzyme decomposition of hydrogen peroxide produces oxonium ion, oxonium ion makes four chloro-naphthols develop the color and precipitate and is positioned enzyme reaction position, sheath fluid sample introduction is to liquid storage tank R7, through the blood sample of chemical staining under the effect of sheath fluid stream, leucocyte is in line and flows into waste liquid pool W1 by detection zone A1 through laser beam irradiation one by one, be classified due to light scattering and cell size difference.Appears as negative reaction without black-and-blue particle in cell after dyeing, occur that the black particle of fine particle or sparse distribution is weak positive reaction, occur that the thick and intensive particle of black is strong positive reaction.
All kinds of leucocyte to the reaction of peroxidase is: early stage myeloblast is for negative, each stage that early young grain is later all contains peroxidase, and strengthen gradually along with its content of maturation of cell, eosinophil has the strongest peroxidase, neutrophil leucocyte contains stronger peroxidase, and basophilic granulocyte is not containing this enzyme; In monocyte system, except the early stage primitive stage, inmature monokaryon and monocyte there will be more weak peroxidase reaction; Lymphocyte, erythroneocytosis, megacaryocyte etc. are all peroxidase negative reaction.To sum up, in blood five kinds of leukocytic peroxidase activities put in order into: eosinophil > neutrophil leucocyte > monocyte, lymphocyte and basophilic granulocyte are all without peroxidase.Due to after enzyme reaction in intensity difference (feminine gender, the weak positive, strong positive) and cell volume difference in size, when cell passes through detection zone A1 through laser beam irradiation, produce the scattered light of varying strength, each cell produces histochemical staining result and light scattering result two signals.
At the scatter diagram that peroxidase detection chip produces, transverse axis represents the cell peroxidase content situation obtained 7 ° of laser light scattering absorptances, and peroxidase strong positive cell is positioned at right-hand member; The longitudinal axis is the cell volume size that low angle scattered light 2.8 ° is measured, and the expression scattered light signal being positioned at top is strong, and cell volume is large.Except basophilic granulocyte, the cell of each type is recorded in specific position according to their feature, eosinophil, neutrophil leucocyte, monocyte and lymphocyte can be distinguished in setting range, and calculate total white blood cells and classification in conjunction with basophilic granulocyte detection chip result.To sum up, cell is divided into five classes by peroxidase detection chip: the 1) eosinophil of the most strong positive of peroxidase; 2) neutrophilic segmented granulocyte of peroxidase strong positive; 3) comparatively large, the p+ monocyte of peroxidase of volume; 4) lymphocyte of small volume, peroxidase feminine gender; 5) volume is greater than lymphocyte and the maxicell that is unstained of peroxidase feminine gender, and this type of cell increases prompting naivety or original various types of cells may occur.
Step 200: adopt biological dissolution technology and electrical impedance method to detect basophilic granulocyte;
In step 200, employing biological dissolution technology and electrical impedance method detect basophilic granulocyte detailed process: anticoagulation sample introduction is to liquid storage tank S2, special basophilic granulocyte dilution sample introduction is to liquid storage tank R3, quantitative anticoagulation and quantitative dilution flow out from respective liquid storage tank and realize mixing, special basophilic granulocyte reagent is by the leucocyte removing cell membrane except basophilic granulocyte, make its bare nucleus and smaller volume, and erythrocytolysis, only basophilic granulocyte keeps original state, volume is obviously greater than the leucocyte of other types, sheath fluid sample introduction is to liquid storage tank R8, treated blood sample is under the effect of quantitative sheath fluid stream, leucocyte is in line and one by one by detection zone A2, A2 upper and lower two ends in detection zone apply steady current, detect (namely each cell produces the electronic impulse proportional with cell volume) through electrical impedance and flow into waste liquid pool W2, according to threshold value setting through signal processing system Treatment Analysis, distinguish leucocyte bare nucleus and basophilic granulocyte, provide total white blood cells and basophilic granulocyte Effective Numerical more accurately.
Step 300: adopt two multi-angle laser method to detect red blood cell and blood platelet, and adopt red blood cell optical scattering method to detect haemoglobin.
In step 300, low angle scattered light (1 °-3 °) is for measuring erythrocyte volume, high angle scattered light (5 °-15 °) measures each endoerythrocytic hemoglobin concentration (MCH) and content of hemoglobin mean value MCHC, and calculates the haemoglobin dispersion of distribution (HDW) value in red blood cell.In embodiments of the present invention, adopt low angle scattered light 2.8 ° for measuring cell volume; High angle scattered light 7 ° measures each endoerythrocytic hemoglobin concentration (MCH) and content of hemoglobin mean value MCHC, and calculates the haemoglobin dispersion of distribution (HDW) value in red blood cell.Platelet count analysis have employed two-dimentional optical triangulation method too, high angle scattered light (5 °-15 °) can measure cell refractive index (RI), low angle scattered light (1 °-3 °) can measure cell size, in embodiments of the present invention, adopt high angle scattered light 7 ° to measure cell refractive index (RI), low angle scattered light 2.8 ° can measure cell size.
The blood cell analysis chip of the embodiment of the present invention, analyser and analytical approach carry out somatotype to haemocyte in anticoagulation, have that structure is simple, volume is little, cost is low, easy to operate, easy care, easily transport, chip mistake and the advantage such as discardable, meet the demand for development of analytical instrument microminiaturization, integrated and portability, be applicable to the uses such as hospital, clinic, community and individual family.
The foregoing is only preferred embodiment of the present invention, not in order to limit the present invention, all any amendments done within the spirit and principles in the present invention, equivalent replacement and improvement etc., all should be included within protection scope of the present invention.
Claims (8)
1. a blood cell analysis chip, it is characterized in that, comprise: leukocyte analysis chip and red blood cell, blood platelet or hemoglobin analysis chip, described leukocyte analysis chip comprises peroxidase detection chip and basophilic granulocyte detection chip, described peroxidase detection chip utilizes laser light scattering and peroxidase staining technique to carry out cell classification, described basophilic granulocyte detection chip adopts biological dissolution and electrical impedance method to detect basophilic granulocyte, described red blood cell, blood platelet or hemoglobin analysis chip adopt two multi-angle laser method to detect red blood cell and blood platelet, and adopt red blood cell optical scattering method to detect haemoglobin,
Peroxidase detection chip is provided with liquid storage tank, waste liquid pool W1, detection zone and multiple dosing assemblies, described liquid storage tank comprises liquid storage tank S1, liquid storage tank R1, liquid storage tank R2, liquid storage tank R5, liquid storage tank R6 and liquid storage tank R7, liquid storage tank S1 is for storing anticoagulation, liquid storage tank R1 is used for store washing liquid, liquid storage tank R2 is for storing formaldehyde isotonic liquid, liquid storage tank R5 is for storing hydrogen peroxide, liquid storage tank R6 is for storing four chloro-naphthols, liquid storage tank R7 is for storing sheath fluid, described peroxidase detection chip is also provided with first fluid passage, second fluid passage, 3rd fluid passage, 4th fluid passage, 5th fluid passage, 6th fluid passage and sense channel, liquid storage tank S1 is communicated with waste liquid pool W1 with sense channel by first fluid passage, liquid storage tank R1 is communicated with waste liquid pool W1 with sense channel by second fluid passage, liquid storage tank R2 is communicated with waste liquid pool W1 with sense channel by the 3rd fluid passage, second fluid passage and the 3rd fluid passage and sense channel cross at same position, liquid storage tank R5 is communicated with waste liquid pool W1 with sense channel by the 4th fluid passage, liquid storage tank R6 is communicated with waste liquid pool W1 with sense channel by the 5th fluid passage, 4th fluid passage and the 5th fluid passage and sense channel cross at same position, liquid storage tank R7 is communicated with waste liquid pool W1 with sense channel by two article of the 6th fluid passage, article two, the 6th fluid passage and sense channel cross at same position, second fluid passage and the 3rd fluid passage are in the position that crosses of sense channel, 4th fluid passage and the 5th fluid passage are in the position that crosses of sense channel, article two, the 6th fluid passage is successively set on sense channel in the position that crosses of sense channel, and second fluid passage and the 3rd fluid passage in the position that crosses of sense channel near liquid storage tank S1, article two, the 6th fluid passage in the position that crosses of sense channel near waste liquid pool W1, multiple dosing assemblies is located at described first fluid passage respectively, second fluid passage, 3rd fluid passage, 4th fluid passage, on 5th fluid passage and the 6th fluid passage, described waste liquid pool is for storing the blood sample through detecting, described detection zone carries out laser beam irradiation for making the leucocyte of blood sample through detection zone, classified by light scattering and the different blood sample to detecting of cell size, described dosing assemblies is used for making corresponding fluids in each liquid storage tank quantitative.
2. blood cell analysis chip according to claim 1, it is characterized in that, described basophilic granulocyte detection chip is provided with liquid storage tank, waste liquid pool, detection zone and dosing assemblies, described liquid storage tank is used for storage of blood sample and dilution, described waste liquid pool is for storing the blood sample through detecting, the cell that described detection zone is used for blood sample adopts electrical impedance technology to detect according to cell size through detection zone, the Effective Numerical of leucocyte bare nucleus and basophilic granulocyte is calculated by electronic impulse size and number, described dosing assemblies is used for making corresponding fluids in each liquid storage tank quantitative.
3. blood cell analysis chip according to claim 1, it is characterized in that, described red blood cell, blood platelet or haemoglobin detection chip are provided with liquid storage tank, waste liquid pool, detection zone and dosing assemblies, described liquid storage tank is used for storage of blood sample and dilution, described waste liquid pool is for storing the blood sample through detecting, described detection zone carries out laser beam irradiation for making the leucocyte of blood sample through detection zone, classified by light scattering and the different blood sample to detecting of cell size, described dosing assemblies is used for making corresponding fluids in each liquid storage tank quantitative.
4. the blood cell analysis chip according to any one of claims 1 to 3, is characterized in that, described leukocyte analysis chip and red blood cell, blood platelet or hemoglobin analysis chip adopt material to comprise quartz, glass, monocrystalline silicon or macromolecule polymeric material; Described liquid storage tank sample introduction adopts Micropump, electrokinetic injection, positive pressure to drive sample introduction, Ngatively pressurized sampling or electric osmose sample introduction various ways.
5. a cellanalyzer, it is characterized in that, comprise blood cell analysis chip as claimed in claim 1, detecting unit and signal processing system, described blood cell analysis chip, detecting unit is connected successively with signal processing system, described blood cell analysis chip is for carrying haemocyte, described detecting unit is used for applying laser beam to detection zone and irradiates haemocyte, described signal processing system applies steady current to upper and lower two ends, detection zone, detect each cell and produce the electronic impulse proportional with cell volume, according to the threshold value of setting, distinguish leucocyte bare nucleus and basophilic granulocyte, provide total white blood cells and basophilic granulocyte Effective Numerical accurately, or described signal processing system adopts high angle scattered light to measure cell peroxidase content situation, endoerythrocytic hemoglobin concentration, content of hemoglobin mean value or measurement cell refractive index, described signal processing system adopts low angle scattered light to measure cell volume or size.
6. adopt blood cell analysis chip as claimed in claim 1 to carry out a blood cell analysis method for blood cell analysis, it is characterized in that, comprising:
Step a: utilize laser light scattering and peroxidase staining technique to classify to cell;
Step b: adopt biological dissolution technology and electrical impedance method to detect basophilic granulocyte;
Step c: adopt two multi-angle laser method to detect red blood cell and blood platelet, and adopt red blood cell optical scattering method to detect haemoglobin;
Described step a comprises: sample introduction is to liquid storage tank S1, R1, R2 respectively for anticoagulation, dilution, clean-out system and formaldehyde isotonic liquid, and whole blood carries out suitably dilution and the short time hatches; Mix and heat with hydrogen peroxide and four chloro-naphthols through liquid storage tank R5, R6 respectively, intracellular peroxidation thing enzyme decomposition of hydrogen peroxide produces oxonium ion, and oxonium ion makes four chloro-naphthols develop the color and precipitate and is positioned enzyme reaction position; Sheath fluid sample introduction is to liquid storage tank R7, and through the blood sample of chemical staining under the effect of sheath fluid stream, leucocyte is in line and successively by detection zone A1; Flow into waste liquid pool W1 through laser beam irradiation, classify according to light scattering and cell size difference.
7. blood cell analysis method according to claim 6, it is characterized in that, described step b comprises: anticoagulation sample introduction is to liquid storage tank S2, special basophilic granulocyte dilution sample introduction is to liquid storage tank R3, quantitative anticoagulation and quantitative dilution flow out from respective liquid storage tank and mix, sheath fluid sample introduction is to liquid storage tank R8, treated blood sample is under the effect of quantitative sheath fluid stream, leucocyte is in line and successively by count detection district A2, A2 upper and lower two ends in detection zone apply steady current, detect through electrical impedance and flow into waste liquid pool W2, according to threshold value setting through signal processing system Treatment Analysis, distinguish leucocyte bare nucleus and basophilic granulocyte, provide total white blood cells and basophilic granulocyte Effective Numerical accurately.
8. blood cell analysis method according to claim 6, it is characterized in that, in described step c, described pair of multi-angle laser method detects to comprise and uses low angle scattered light and high angle scattered light, and wherein, described low angle scattered light is for measuring erythrocyte volume, high angle scattered light measures each endoerythrocytic hemoglobin concentration and content of hemoglobin mean value, on platelet count is analyzed, high angle scattering optical power measurement cell refractive index, low angle scattering optical power measurement cell size.
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| CN106845622B (en) * | 2017-01-22 | 2019-04-23 | 江西特康科技有限公司 | Platelet count method and system |
| CN110226083B (en) | 2017-02-17 | 2022-12-20 | 深圳迈瑞生物医疗电子股份有限公司 | Erythrocyte fragment recognition method and device, blood cell analyzer and analysis method |
| CN107219195B (en) * | 2017-05-23 | 2019-07-23 | 山东中医药大学附属医院 | A kind of blood leucocyte detection device and method |
| WO2019206310A1 (en) | 2018-04-28 | 2019-10-31 | 深圳迈瑞生物医疗电子股份有限公司 | Blood analysis method, blood analysis system, and storage medium |
| CN111656161B (en) * | 2018-04-28 | 2024-02-20 | 深圳迈瑞生物医疗电子股份有限公司 | Sample analyzer abnormality alarming method, system and storage medium |
| CN111801568B (en) * | 2018-04-28 | 2024-05-24 | 深圳迈瑞生物医疗电子股份有限公司 | Method and system for measuring platelet concentration |
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