CN103472034A - Blood cell analysis chip, analysis meter and analysis method - Google Patents
Blood cell analysis chip, analysis meter 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, analysis meter and analysis method. The blood cell analysis chip provided by the invention comprises a white blood cell analysis chip, a red blood cell, blood platelet or hemoglobin analysis chip, wherein the white blood cell analysis chip comprises a peroxidase detection chip and a basophilic granulocyte detection chip; the peroxidase detection chip performs cell classification by utilizing laser scattering and peroxidase staining technology; the basophilic granulocyte detection chip detects basophilic granulocytes by adopting the biological dissolving and electrical impedance method; the red blood cell, blood platelet or hemoglobin analysis chip detects red blood cells and blood platelets by adopting the double-angle laser method, and detects hemoglobin by adopting the red blood cell optical scattering method. The blood cell analysis chip disclosed by the invention has the advantages that the structure is simple, the size is small, the cost is low, the operation is convenient, the maintenance operation is easy, the easy transportation is realized, the chip can be discarded after being used, and the like.
Description
Technical field
The invention belongs to the blood cell analysis technical field, relate in particular to a kind of blood cell analysis chip, analysis and analytical approach.
Background technology
The information that blood cell analysis obtains can contribute to diagnosis, the antidiastole disease 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 check (being routine blood test) becomes first of three large routine inspections in clinical examination (routine blood test, routine urinalysis, just conventional), 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 analysers are come out one after another, and the blood cell analysis technology is developed rapidly.
For blood cell analysis, generally by commercially available cellanalyzer, through the specific people, carry out under given conditions at present.Existing cellanalyzer is bulky, expensive, complicated operation, need the 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 more concentrated hospital inspection section office of test samples, for different medical units such as village's clinics, often sample size is little, and sample disperses very much on time dimension, the situation that significant discomfort occurs, therefore should overcome tradition check kind equipment shortcoming, meet again the demand of different medical unit to measuring means, be badly in need of developing at present portability, simple to operateization, the cellanalyzer of the 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, the technical matters that needs the special messenger to use and carry out 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 angle laser methods 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: be provided with liquid storage tank, waste liquid pool, detection zone and quantitative assembly on described peroxidase detection chip, 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 while for the leucocyte that makes blood sample, passing through detection zone, by light scattering and cell size difference, the blood sample detected is classified, described quantitative assembly is for making the interior corresponding fluids of each liquid storage tank quantitative.
Technical scheme of the present invention also comprises: be provided with liquid storage tank, waste liquid pool, detection zone and quantitative assembly on described basophilic granulocyte detection chip, described liquid storage tank is for storage of blood sample and dilution, described waste liquid pool is for storing the blood sample through detecting, described detection zone adopts electrical impedance technology to be detected according to cell size for the cell of blood sample during through detection zone, calculate the Effective Numerical of leucocyte bare nucleus and basophilic granulocyte by electronic impulse size and number, described quantitative assembly is for making the interior corresponding fluids of each liquid storage tank quantitative.
Technical scheme of the present invention also comprises: be provided with liquid storage tank, waste liquid pool, detection zone and quantitative assembly on described red blood cell, blood platelet or haemoglobin detection chip, described liquid storage tank is 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 while for the leucocyte that makes blood sample, passing through detection zone, by light scattering and cell size difference, the blood sample detected is classified, described quantitative assembly is for making the interior corresponding fluids of 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 irradiated haemocyte for detection zone is applied to laser beam, described signal processing system applies steady current to the upper and lower two ends of detection zone, detect each cell and produce electronic impulse proportional to 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 the 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 the 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 comprises:
Step a: utilize laser light scattering and peroxidase staining technique to be classified to cell;
Step b: adopt biological dissolution technology and electrical impedance method to detect basophilic granulocyte;
Step c: adopt two angle laser methods 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 tetrachloro-naphthols respectively through liquid storage tank R5, R6, cell endoperoxide enzyme decomposition of hydrogen peroxide produces oxonium ion, and oxonium ion develops the color tetrachloro-naphthols and precipitate to be positioned the enzyme reaction position; The 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, classified according to light scattering and cell size difference.
Technical scheme of the present invention also comprises: described step b comprises: the anticoagulation sample introduction is to liquid storage tank S2, special basophilic granulocyte dilution sample introduction is to liquid storage tank R3, quantitatively anticoagulation and quantitatively dilution flow out and mix from liquid storage tank separately, the sheath fluid sample introduction is to liquid storage tank R8, treated blood sample is under quantitative sheath fluid stream effect, leucocyte is in line and passes through successively detection zone A2, the upper and lower two ends of detection zone A2 apply steady current, detect and flow into waste liquid pool W2 through electrical impedance, according to Threshold through the 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 angle laser method detects and comprises use low angle scattered light and high angle scattered light, wherein, described low angle scattered light is for measuring erythrocyte volume, the high angle scattered light is measured 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 of the present invention has following advantage or beneficial effect: blood cell analysis chip, analyser and the analytical approach of the embodiment of the present invention are carried out somatotype to haemocyte in anticoagulation, have simple in structure, volume is little, cost is low, easy to operate, easy care, easily transportation, chip are the advantage such as discardable by mistake, the demand for development that meets analytical instrument microminiaturization, integrated and portability, be applicable to the uses such as hospital, clinic, community and individual family.
The 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 peroxidase detection chip of the blood cell analysis chip of the embodiment of the present invention;
Accompanying drawing 3 is structural representations of basophilic granulocyte detection chip of the blood cell analysis chip of the embodiment of the present invention;
Accompanying drawing 4 is structural representations of red blood cell, blood platelet or the haemoglobin detection chip of the blood cell analysis chip of the embodiment of the present invention;
Accompanying drawing 5 is structural representations of the cellanalyzer of the embodiment of the present invention;
Accompanying drawing 6 is concrete application structure schematic diagram 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 purpose of the present invention, technical scheme and advantage clearer, 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, is 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, the leukocyte analysis chip comprises: peroxidase detection chip and basophilic granulocyte detection chip.Refer to Fig. 2, Fig. 2 is the structural representation of peroxidase detection chip of the blood cell analysis chip of the embodiment of the present invention.Be provided with liquid storage tank, waste liquid pool, detection zone and quantitative assembly on the peroxidase detection chip.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 while for the leucocyte that makes blood sample, passing through detection zone, thereby by light scattering and the different blood samples to detection of cell size, classified, quantitatively assembly is for making the interior corresponding fluids of each liquid storage tank quantitative.In embodiments of the present invention, liquid storage tank arranges at least one, its can be used for the storage cell is cleaned and is destroyed detection reagent, make cell carry out the zymolytic detection reagent of superoxide and sheath fluid.The peroxidase detection chip utilizes laser light scattering and peroxidase staining technique to carry out cell classification, the peroxidase detection chip is carried out the 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 of clinical or Physiological Experiment use, its osmotic pressure equates with plasma osmolarity is called isotonic solution, whole blood carries out suitably dilution and the short time hatches, now cell is cleaned agent destruction, leucocyte slurry endoenzyme is fixed, by liquid storage tank R5, R6 mixes and heats with hydrogen peroxide and tetrachloro-naphthols respectively, now cell endoperoxide enzyme decomposition of hydrogen peroxide produces oxonium ion, oxonium ion develops the color tetrachloro-naphthols and precipitates to be positioned the enzyme reaction position, the sheath fluid sample introduction is to the 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, because light scattering and cell size are different, be classified, quantitatively assembly C1 to C7 is for making in each liquid storage tank corresponding fluids quantitative.Appear 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.The reaction of all kinds of leucocytes to peroxidase: early stage myeloblast is negative, early the children is contained peroxidase at each later stage of grain, and along with its content of maturation of cell strengthens gradually, eosinophil has the strongest peroxidase, neutrophil leucocyte contains stronger peroxidase, and basophilic granulocyte is not containing this enzyme; In the monocyte system, except the early stage primitive stage, inmature monokaryon and monocyte there will be weak peroxidase reaction; Lymphocyte, inmature red blood cell, megacaryocyte etc. are all the peroxidase negative reaction.To sum up, in blood, five kinds of leukocytic peroxidase activities put in order as eosinophil > neutrophil leucocyte > monocyte, lymphocyte and basophilic granulocyte are all without peroxidase.Owing to being intensity difference (feminine gender, the weak positive, strong positive) and cell volume difference in size after enzyme reaction, when cell passes through detection zone A1 through laser beam irradiation, produce scattered light, each cell generation group coloration result and two signals of light scattering result of varying strength.
The scatter diagram produced in the peroxidase detection chip, transverse axis is illustrated in the cell peroxidase content situation that 7 ° of laser light scattering absorptances obtain, and peroxidase strong positive cell is positioned at right-hand member; The longitudinal axis is the cell volume size of 2.8 ° of measurements of low angle scattered light, and the expression scattered light signal that is 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 characteristics, 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, the peroxidase detection chip is divided into five classes by cell: the 1) eosinophil of the strong positive of peroxidase; 2) neutrophilic segmented granulocyte of peroxidase strong positive; 3) large, the p+ monocyte of peroxidase of volume; 4) lymphocyte of small volume, peroxidase feminine gender; 5) volume is greater than the maxicell that is unstained of lymphocyte and peroxidase feminine gender, and this type of cell increases the 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.Be provided with liquid storage tank, waste liquid pool, detection zone and quantitative assembly on the basophilic granulocyte detection chip of the embodiment of the present invention.Liquid storage tank is 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 embodiment of the present invention, basophilic granulocyte detection chip used diluent is available reagent.The detailed process that the basophilic granulocyte detection chip is detected is: the anticoagulation sample introduction is to the S2 liquid storage tank, special basophilic granulocyte dilution sample introduction is to the R3 liquid storage tank, quantitatively anticoagulation and quantitatively dilution flow out and realize mixing from liquid storage tank separately, special-purpose basophilic granulocyte reagent is removed cell membrane by the leucocyte except basophilic granulocyte, make its bare nucleus smaller volume, and erythrocytolysis, only basophilic granulocyte keeps original state, volume obviously is greater than the leucocyte of other types, the sheath fluid sample introduction is to the R8 liquid storage tank, treated blood sample is under quantitative sheath fluid stream effect, leucocyte is in line and passes through one by one detection zone A2, the upper and lower two ends of detection zone A2 apply steady current, detect (being that each cell produces electronic impulse proportional to cell volume) and flow into waste liquid pool W2 through electrical impedance, according to Threshold through the 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 embodiment of the present invention, the basophilic granulocyte detection chip adopts biological dissolution technology and electrical impedance method to detect basophilic granulocyte, further improve and detect performance, can also adopt optical detection to be detected, quantitatively assembly C8 to C11 is for making in each liquid storage tank corresponding fluids quantitative.
Referring to Fig. 4, is the structural representation of red blood cell, blood platelet or the haemoglobin detection chip of the blood cell analysis chip of the embodiment of the present invention.Be provided with liquid storage tank, waste liquid pool, detection zone and quantitative assembly on red blood cell, blood platelet or haemoglobin detection chip.Liquid storage tank is for storage of blood sample and dilution, and waste liquid pool is for storing the blood sample through detecting, and detection zone carries out Ear Mucosa Treated by He Ne Laser Irradiation while being used for making red blood cell by 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 angle laser methods, microcyte, bib and blood platelet are more easily differentiated, greatly improve accuracy and precision that blood platelet is measured, and adopt the direct Measuring hemoglobin of red blood cell scattered light.The detailed process that the red analysis chip of red blood cell and blood platelet is detected is: the anticoagulation sample introduction is to liquid storage tank S3, special dilution sample introduction is to liquid storage tank R4, quantitatively anticoagulation and quantitatively dilution flow out and realize mixing from liquid storage tank separately, the sheath fluid sample introduction is to liquid storage tank R9, treated blood sample is under quantitative sheath fluid stream effect, red blood cell is in line and flows into waste liquid pool W3 by detection zone A3 one by one, Ear Mucosa Treated by He Ne Laser Irradiation scanning is by each red blood cell of detection zone A3, and 2.8 ° of low angle scattered lights are for measuring cell volume; 7 ° of each endoerythrocytic hemoglobin concentration (MCH) of measurement of high angle scattered light and content of hemoglobin mean value MCHC, and calculate the haemoglobin dispersion of distribution (HDW) value in red blood cell.Adopted too two-dimentional light-scattering analysis method in the platelet count analysis, 7 ° of high angle scattered lights can be measured cell refractive index (RI), 2.8 ° of low angle scattered lights can be measured cell size, draw platelet counts and correlation parameter on two-dimensional scattering figure, can measure blood platelet of all sizes in 1-60fl, also can be measured blood platelet content content.Quantitatively assembly C12 to C15 is for making in each liquid storage tank corresponding fluids 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 be by quartz, glass, monocrystalline silicon, macromolecule polymeric material as making such as polymetylmethacrylate, polydimethylsiloxane, polycarbonates; In addition, in order to alleviate or to avoid the chip microchannel surface to adsorb haemocyte, the chip microchannel inside surface can be through the ad hoc fashion modification, or the 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 and signal processing system are connected successively, the blood cell analysis chip is for carrying haemocyte, detecting unit is irradiated haemocyte for detection zone is applied to laser beam, signal processing system applies steady current to the upper and lower two ends of aperture, detect each cell and produce electronic impulse proportional to 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 measurement cell peroxidase content situation, endoerythrocytic hemoglobin concentration, content of hemoglobin mean value or measures the cell refractive index, described signal processing system employing low angle scattered light measurement cell volume or size.
Referring to Fig. 6, is the concrete application structure schematic diagram 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 the described blood cell analysis chip of Fig. 1, for carrying haemocyte 8, light source 1 shines 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 all are connected with diaphragm 5 and photodetector 6.The cellanalyzer of the embodiment of the present invention is except being applied to medical industry, in every diameter that relates to requirement measurement of species particle and liquid, atomic quantity carries out quantitatively and qualitatively analyzing, can measure with this instrument, be that this instrument can develop into laser particle size analyzer, for example, 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 etc., to microelectronic component, and the production of integrated circuit, quality control, pharmaceutical industry and chemical industry, Food Hygiene Surveillances 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 be classified to cell;
In step 100, utilize laser light scattering and peroxidase staining technique to classify and be specially cell: 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 cleaned agent destruction, leucocyte slurry endoenzyme is fixed, by liquid storage tank R5, R6 mixes and heats with hydrogen peroxide and tetrachloro-naphthols respectively, now cell endoperoxide enzyme decomposition of hydrogen peroxide produces oxonium ion, oxonium ion develops the color tetrachloro-naphthols and precipitates to be positioned the enzyme reaction position, the 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, because light scattering and cell size are different, be classified.Appear 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 leucocytes to the reaction of peroxidase are: early stage myeloblast is negative, early the children is contained peroxidase at each later stage of grain, and along with its content of maturation of cell strengthens gradually, eosinophil has the strongest peroxidase, neutrophil leucocyte contains stronger peroxidase, and basophilic granulocyte is not containing this enzyme; In the monocyte system, except the early stage primitive stage, inmature monokaryon and monocyte there will be weak peroxidase reaction; Lymphocyte, inmature red blood cell, megacaryocyte etc. are all the peroxidase negative reaction.To sum up, in blood, five kinds of leukocytic peroxidase activities put in order as eosinophil > neutrophil leucocyte > monocyte, lymphocyte and basophilic granulocyte are all without peroxidase.Owing to being intensity difference (feminine gender, the weak positive, strong positive) and cell volume difference in size after enzyme reaction, when cell passes through detection zone A1 through laser beam irradiation, produce scattered light, each cell generation group coloration result and two signals of light scattering result of varying strength.
The scatter diagram produced in the peroxidase detection chip, transverse axis is illustrated in the cell peroxidase content situation that 7 ° of laser light scattering absorptances obtain, and peroxidase strong positive cell is positioned at right-hand member; The longitudinal axis is the cell volume size of 2.8 ° of measurements of low angle scattered light, and the expression scattered light signal that is 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 characteristics, 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, the peroxidase detection chip is divided into five classes by cell: the 1) eosinophil of the strong positive of peroxidase; 2) neutrophilic segmented granulocyte of peroxidase strong positive; 3) large, the p+ monocyte of peroxidase of volume; 4) lymphocyte of small volume, peroxidase feminine gender; 5) volume is greater than the maxicell that is unstained of lymphocyte and peroxidase feminine gender, and this type of cell increases the 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, adopt biological dissolution technology and electrical impedance method detection basophilic granulocyte detailed process to be: the anticoagulation sample introduction is to liquid storage tank S2, special basophilic granulocyte dilution sample introduction is to liquid storage tank R3, quantitatively anticoagulation and quantitatively dilution flow out and realize mixing from liquid storage tank separately, special-purpose basophilic granulocyte reagent is removed cell membrane by the leucocyte except basophilic granulocyte, make its bare nucleus smaller volume, and erythrocytolysis, only basophilic granulocyte keeps original state, volume obviously is greater than the leucocyte of other types, the sheath fluid sample introduction is to liquid storage tank R8, treated blood sample is under quantitative sheath fluid stream effect, leucocyte is in line and passes through one by one detection zone A2, the upper and lower two ends of detection zone A2 apply steady current, detect (being that each cell produces electronic impulse proportional to cell volume) and flow into waste liquid pool W2 through electrical impedance, according to Threshold through the 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 angle laser methods 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 °) is measured 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 embodiment of the present invention, adopt 2.8 ° of low angle scattered lights for measuring cell volume; 7 ° of each endoerythrocytic hemoglobin concentration (MCH) of measurement of high angle scattered light and content of hemoglobin mean value MCHC, and calculate the haemoglobin dispersion of distribution (HDW) value in red blood cell.Adopted too two-dimentional light-scattering analysis method in the platelet count analysis, high angle scattered light (5 °-15 °) can be measured cell refractive index (RI), low angle scattered light (1 °-3 °) can be measured cell size, in embodiment of the present invention, adopt 7 ° of high angle scattered lights to measure cell refractive indexes (RI), 2.8 ° of low angle scattered lights can be measured cell size.
Blood cell analysis chip, analyser and the analytical approach of the embodiment of the present invention are carried out somatotype to haemocyte in anticoagulation, have simple in structure, volume is little, cost is low, easy to operate, easy care, easily transportation, chip are the advantage such as discardable by mistake, the demand for development that meets 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 modifications of doing within the spirit and principles in the present invention, be equal to and replace and improvement etc., within all should being included in protection scope of the present invention.
Claims (10)
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 angle laser methods to detect red blood cell and blood platelet, and adopt red blood cell optical scattering method to detect haemoglobin.
2. blood cell analysis chip according to claim 1, it is characterized in that, be provided with liquid storage tank, waste liquid pool, detection zone and quantitative assembly on described peroxidase detection chip, 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 while for the leucocyte that makes blood sample, passing through detection zone, by light scattering and cell size difference, the blood sample detected is classified, described quantitative assembly is for making the interior corresponding fluids of each liquid storage tank quantitative.
3. blood cell analysis chip according to claim 1, it is characterized in that, be provided with liquid storage tank on described basophilic granulocyte detection chip, waste liquid pool, detection zone and quantitative assembly, described liquid storage tank is for storage of blood sample and dilution, described waste liquid pool is for storing the blood sample through detecting, described detection zone adopts electrical impedance technology to be detected according to cell size for the cell of blood sample during through detection zone, calculate the Effective Numerical of leucocyte bare nucleus and basophilic granulocyte by electronic impulse size and number, described quantitative assembly is for making the interior corresponding fluids of each liquid storage tank quantitative.
4. blood cell analysis chip according to claim 1, it is characterized in that, described red blood cell, be provided with liquid storage tank on blood platelet or haemoglobin detection chip, waste liquid pool, detection zone and quantitative assembly, described liquid storage tank is 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 while for the leucocyte that makes blood sample, passing through detection zone, by light scattering and cell size difference, the blood sample detected is classified, described quantitative assembly is for making the interior corresponding fluids of each liquid storage tank quantitative.
5. according to the described blood cell analysis chip of claim 2 to 4 any one, it 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.
6. 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 irradiated haemocyte for detection zone is applied to laser beam, described signal processing system applies steady current to the upper and lower two ends of detection zone, detect each cell and produce electronic impulse proportional to 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 the 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 the low angle scattered light to measure cell volume or size.
7. a blood cell analysis method, is characterized in that, comprising:
Step a: utilize laser light scattering and peroxidase staining technique to be classified to cell;
Step b: adopt biological dissolution technology and electrical impedance method to detect basophilic granulocyte;
Step c: adopt two angle laser methods to detect red blood cell and blood platelet, and adopt red blood cell optical scattering method to detect haemoglobin.
8. blood cell analysis method according to claim 7, is characterized in that, 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 tetrachloro-naphthols respectively through liquid storage tank R5, R6, cell endoperoxide enzyme decomposition of hydrogen peroxide produces oxonium ion, and oxonium ion develops the color tetrachloro-naphthols and precipitate to be positioned the enzyme reaction position; The 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, classified according to light scattering and cell size difference.
9. blood cell analysis method according to claim 7, it is characterized in that, described step b comprises: the anticoagulation sample introduction is to liquid storage tank S2, special basophilic granulocyte dilution sample introduction is to liquid storage tank R3, quantitatively anticoagulation and quantitatively dilution flow out and mix from liquid storage tank separately, the sheath fluid sample introduction is to liquid storage tank R8, treated blood sample is under quantitative sheath fluid stream effect, leucocyte is in line and passes through successively count detection district A2, the upper and lower two ends of detection zone A2 apply steady current, detect and flow into waste liquid pool W2 through electrical impedance, according to Threshold through the signal processing system Treatment Analysis, distinguish leucocyte bare nucleus and basophilic granulocyte, provide total white blood cells and basophilic granulocyte Effective Numerical accurately.
10. blood cell analysis method according to claim 7, it is characterized in that, in described step c, described pair of angle laser method detects and comprises use low angle scattered light and high angle scattered light, and wherein, described low angle scattered light is for measuring erythrocyte volume, the high angle scattered light is measured 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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