CN103471980B - A kind of chip type blood cell analyzer and method - Google Patents
A kind of chip type blood cell analyzer and method Download PDFInfo
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- CN103471980B CN103471980B CN201310373222.3A CN201310373222A CN103471980B CN 103471980 B CN103471980 B CN 103471980B CN 201310373222 A CN201310373222 A CN 201310373222A CN 103471980 B CN103471980 B CN 103471980B
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Abstract
The invention belongs to blood cell analysis technical field, particularly relate to a kind of chip type blood cell analyzer and method.Chip type blood cell analyzer of the present invention comprises blood cell analysis chip, institute's blood cell analysis chip comprises leucocyte/hemoglobin analysis chip and red blood cell/plaque assay chip, described leucocyte/hemoglobin analysis chip adopts electrical impedance, high frequency conductance and laser light scattering detection technique to detect leucocyte, and described red blood cell/plaque assay chip adopts sheath Flow Technique, electrical impedance technology or floating landmark technique to count respectively red blood cell and blood platelet.The somatotype of the present invention by utilizing blood cell analysis chip to realize haemocyte, 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 particularly useful for the use of Site Detection, emergency analysis, domestic. applications and different medical unit.
Description
Technical field
The invention belongs to blood cell analysis technical field, particularly relate to a kind of chip type blood cell analyzer and method.
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, traditional cellanalyzer is bulky, expensive and 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, and 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, do not meet analytical instrument microminiaturization, the demand for development of integrated and portability, and can not village health posts be met, clinic, community or individual family etc. are compared with the user demand of subsection.Therefore conventional inspection kind equipment shortcoming should be overcome, meet the demand of different medical unit to measuring means again, current urgent need develops the cellanalyzer of portability, simple operation, just-in-time of reporting the result, and is applicable to Site Detection, emergency analysis, domestic. applications and primary care.
Summary of the invention
The invention provides a kind of chip type blood cell analyzer and method, be intended to solve bulky, the expensive and complicated operation of existing cellanalyzer, the technical matters of the user demand of less medical institutions can not be met.
Technical scheme provided by the invention is: a kind of chip type blood cell analyzer, comprise blood cell analysis chip, described blood cell analysis chip comprises leucocyte/hemoglobin analysis chip and red blood cell/plaque assay chip, described leucocyte/hemoglobin analysis chip adopts electrical impedance, high frequency conductance and laser light scattering detection technique to detect leucocyte, and described red blood cell/plaque assay chip adopts sheath Flow Technique, electrical impedance technology or floating landmark technique to count respectively red blood cell and blood platelet.
Technical scheme of the present invention also comprises: described leucocyte/hemoglobin analysis chip adopts described impedance bioelectrical measurement leucocyte volume; Adopt described high frequency conductance technology reacting cells internal structural information; Adopt described laser scattering technology analysis of cells particle information.
Technical scheme of the present invention also comprises: described leucocyte/hemoglobin analysis chip adopts the concentration of colorimetric determination haemoglobin; The detection mode of the concentration of described detection haemoglobin is: colorimetric under the specific wavelength of 530-550nm also measures the absorbance of blood sample.
Technical scheme of the present invention also comprises: described leucocyte/hemoglobin analysis chip is provided with liquid storage tank, waste liquid pool, detection zone, colorimetric pool and flow sensor, described liquid storage tank is for storing detection reagent, described waste liquid pool is for storing the blood sample through detecting, described detection zone is used for detecting the leucocyte of process, determine each subgroup character of leucocyte, described colorimetric pool is for detecting the concentration of haemoglobin in blood sample, and described flow sensor is used for making corresponding fluids in liquid storage tank quantitative.
Technical scheme of the present invention also comprises: described red blood cell/plaque assay chip is provided with liquid storage tank, waste liquid pool, detection zone and flow sensor, described liquid storage tank is for storing detection reagent, described waste liquid pool is for storing the blood sample through detecting, the two ends up and down of described detection zone apply steady current, cell through electrical impedance and process produces electronic impulse, height according to pulse counts respectively to red blood cell and blood platelet, and described flow sensor is used for making corresponding fluids in liquid storage tank quantitative; Wherein, described red blood cell and enumeration of thrombocytes mode are: determine cell volume by the measurement of impulse magnitude, obtain Cytometric result by the number of recording impulse; Difference according to blood platelet and erythrocyte volume sets threshold value, pulse signal higher than threshold value is defined as red blood cell, pulse signal lower than threshold value is defined as blood platelet, passes through the number of produced electronic impulse and size and carry out red blood cell and enumeration of thrombocytes and measure analysis.
Technical scheme of the present invention also comprises: described liquid storage tank input mode comprises Micropump, electrokinetic injection, positive pressure driving sample introduction, Ngatively pressurized sampling or electric osmose sample introduction.
Technical scheme of the present invention also comprises: described blood cell analysis chip material comprises quartz, glass, monocrystalline silicon or macromolecule polymeric material.
Another technical scheme provided by the invention is: a kind of chip type blood cell analysis method, comprising:
Step a: adopt electrical impedance, high frequency conductance and laser light scattering detection technique to detect the leucocyte on leucocyte/hemoglobin analysis chip;
Step b: the concentration adopting colorimetric determination haemoglobin;
Step c: adopt sheath Flow Technique, electrical impedance technology or floating landmark technique to count respectively the red blood cell on red blood cell/plaque assay chip and blood platelet.
Technical scheme of the present invention also comprises: in described a, described white blood cell detection mode comprises: adopt described impedance bioelectrical measurement leucocyte volume, adopt described high frequency conductance technology reacting cells internal structural information, adopt described laser scattering technology analysis of cells particle information.
Technical scheme of the present invention also comprises: in described step b, and the detection mode of the concentration of described detection haemoglobin is: colorimetric under the specific wavelength of 530-550nm also measures the absorbance of blood sample.
Technical scheme tool of the present invention has the following advantages or beneficial effect: technical scheme tool of the present invention has the following advantages or beneficial effect: the chip type blood cell analyzer of the embodiment of the present invention and the somatotype of method by utilizing blood cell analysis chip to realize haemocyte, 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 particularly useful for the use of Site Detection, emergency analysis, domestic. applications and different medical unit.
Accompanying drawing explanation
Accompanying drawing 1 is the structural representation of the chip type blood cell analyzer of the embodiment of the present invention;
The detection schematic diagram of the leucocyte/hemoglobin analysis chip of accompanying drawing 2 embodiment of the present invention;
The detection schematic diagram of the red blood cell/plaque assay chip of accompanying drawing 3 embodiment of the present invention;
Accompanying drawing 4 is process flow diagrams of the chip type blood cell analysis method of the embodiment of the present invention;
Accompanying drawing 5 is process flow diagrams of the white blood cell detection 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 chip type blood cell analyzer of the embodiment of the present invention.The chip type blood cell analyzer of the embodiment of the present invention comprises blood cell analysis chip, and blood cell analysis chip comprises leucocyte/hemoglobin analysis chip and red blood cell/plaque assay chip; Wherein,
Leucocyte/hemoglobin analysis chip detects leucocyte for adopting the physics detection techniques such as electrical impedance, high frequency conductance and laser light scattering, particularly,
Seeing also Fig. 2, is the detection schematic diagram of the leucocyte/hemoglobin analysis chip of the embodiment of the present invention.Leucocyte/hemoglobin analysis chip is provided with liquid storage tank, waste liquid pool, detection zone, colorimetric pool and flow sensor, liquid storage tank is for storing detection reagent, waste liquid pool is for storing the blood sample through detecting, detection zone is used for carrying out one by one, simultaneously, triple VCS(volume, conductivity, scatter to the leucocyte of process, V-low frequency wave, analysis of cells volume; C-high frequency waves, analysis of cells caryogram; S-laser, the particle characteristics of analysis of cells; VCS detects and namely uses V, C, S tri-kinds of probes, in a certain site of detection zone, to the single-row leucocyte passed through carry out one by one, simultaneously, triple detections and three dimensional analysis, to determine its subgroup character) detect, and by the determination data each subgroup character of machine process determination leucocyte as calculated, colorimetric pool is for detecting the concentration of haemoglobin, and flow sensor is used for making corresponding fluids in each liquid storage tank quantitative.Wherein, the quantity of liquid storage tank and flow sensor can be arranged according to detection demand, testing process is: under the quantitative effect of flow sensor C1 to C7, by anticoagulation (being the most frequently used in visiting work and one of most important anti-coagulants and reagent), dilution (has certain pH, appropriate osmotic pressure and the balanced electrolyte solution of conductivity, dilute by a certain percentage with human body haemocyte, human body haemocyte can be made to keep normal physiological form, and dispersion is not easily reunited) and hemolytic agent (haemoglobin detected components and erythrocyte hemolysis agent, hemolytic agent is hypotonic, acid, solution containing appropriate wetting agent, red blood cell in the rapid dissolved destruction blood of energy, release haemoglobin also forms with it stable compound) distinguish sample introduction to liquid storage tank S1, R1, R2, whole blood suitably dilutes, and because of hemolytic agent osmotic pressure official post erythrocytolysis destruction, add leucocyte stabilizing agent by liquid storage tank R5 (a kind ofly highly to ooze, the solution of alkalescence, in energy and hemolytic agent, stabilized leukocyte make it to keep or close to original physiologic form) in and the effect of hemolytic agent, make leukocyte surface, endochylema and cell volume return to ortho states substantially, sheath fluid sample introduction is to liquid storage tank R6, blood sample is under the effect of sheath fluid stream, leucocyte is in line and one by one by detection zone A1, carry out one by one to the leucocyte of process, simultaneously, triple VCS detects (V-low frequency wave, analysis of cells volume, C-high frequency waves, analysis of cells caryogram, S-laser, the particle characteristics of analysis of cells, VCS detects and namely uses V, C, S tri-kinds of probes, in a certain site of detection zone, to the single-row leucocyte passed through carry out one by one, simultaneously, triple detections and three dimensional analysis, to determine its subgroup character), and by determination data as calculated machine process to determine each subgroup character of leucocyte.Because different classes of cell can present obvious difference in information characteristic such as volume, surface characteristics, inner structures, these information characteristic are defined in the three-dimensional scatter diagram formed for three-dimensional coordinate with VCS, five class leucocytes can form specific cell mass in three dimensions, accounting for the number percent of total white blood cells by calculating certain group of cell quantities, five leukocyte differential count results can be obtained; Simultaneously, due to RBC(Red Blood Cell, red blood cell count(RBC), refer to mean constant of red blood cell contained in unit volume blood) discharged haemoglobin by dissolved destruction, haemoglobin and hemolytic agent react and form stable haemoglobin compound, haemoglobin compound enters haemoglobin test macro-colorimetric pool E, colorimetric under the specific wavelength of 530-550nm also measures its absorbance, in the change of absorbance and liquid, the content of haemoglobin is proportional, can record the concentration of haemoglobin thus; In embodiments of the present invention, to leukocytic detection except five leukocyte differential counts, can also measure and analyze the parameters such as the volume of neutrophil leucocyte, caryoplasm ratio, cell granulations characteristic, for understanding the difference of bacterium perception disease and Other diseases, also there is granulophilocyte analytic function and t lymphocyte subset cluster analysis function.
Electrical impedance is detected as measures leucocyte volume according to Ku Erte theory of electrical impedance, measuring method is: certain low-frequency current is by two electrodes of sensor, because cell is non-conducting, when cell is by two electrodes, inter-electrode impedance increases instantaneously, the electric pulse that formation amplitude is directly proportional to cell volume, measures the volume of cell according to the size of pulse, the number according to pulse carries out cell count; Through to various cell produce the electronic selection of impulse magnitude, can the different types of cell of incomplete differentiation, the comparatively significant lymphocyte of effective differentiation volume difference in size and monocyte, but because pulse height that the is dissimilar but cell that volume is identical generation is identical, therefore only can not distinguish completely by cell volume, and then utilize high-frequency electrical inducing defecation by enema and suppository and laser light scattering analysis leucocyte inner structure.
High frequency conductance technology is used for reacting cells internal structural information, and concrete reactive mode is: although cell membrane can not make low-frequency current pass through, by high-frequency current; Electrode adds radio frequency (high frequency) generator, cell membrane has conduction to high-frequency current, when electric current forms alternating electromagnetic field around by during cell, form electromagnetic wave, because the conduction parameter of difference to high-frequency current between the ratio of nucleus, slurry, the inner structure such as particle and nucleocytoplasmic ratio is distinguished to some extent, the electromagnetic field formed around it is not identical yet, can reflect nuclear internal structural information by cell to the conduction parameter of high-frequency current and the variable quantity of electromagnetic field; And leaflet core and non-segmented cell can be distinguished, for distinguishing the cell colony that volume is close and inner structure is different, as lymphocyte and basophilic granulocyte.
Laser scattering technology is used for analysis of cells particle information: concrete analysis mode is: scanned each cell by one-wavelength laser, analysis of cells film surface characteristics and inner structure, detect grain pattern and density case in intracellular nucleic leaflet situation and endochylema, detect the light scattering characteristic of cell 10 ° of-70 ° of wide range, and distinguish the different cell colony of particle characteristics according to the light scattering characteristic of cell, the scattered light thinner than particle that such as intracellular granular is thick is stronger, distinguish the neutrophilia in granulocyte whereby, acidophilia and basophilla three kinds of cells.
Red blood cell/plaque assay chip counts red blood cell and blood platelet respectively for adopting sheath Flow Technique, electrical impedance technology or floating landmark technique; Seeing also Fig. 3, is the detection schematic diagram of the red blood cell/plaque assay chip of the embodiment of the present invention.Red blood cell/plaque assay chip is provided with liquid storage tank, waste liquid pool, detection zone and flow sensor, liquid storage tank is for storing detection reagent, waste liquid pool is for storing the blood sample through detecting, the two ends up and down of detection zone apply steady current, through electrical impedance technology detect make through cell produce electronic impulse, height according to pulse counts respectively to red blood cell and blood platelet, and flow sensor is used for making corresponding fluids in each liquid storage tank quantitative; Wherein, the quantity of liquid storage tank and flow sensor can be arranged according to detection demand.Concrete testing process is: under the quantitative effect of flow sensor C8 to C13, anticoagulation sample introduction is to liquid storage tank S2, dilution sample introduction is to liquid storage tank R5, quantitative whole blood sample dilutes by proper proportion through quantitative dilution, first sheath fluid (dilution) sample introduction is to R6 liquid storage tank, second sheath fluid (dilution) sample introduction is to R7 liquid storage tank, blood sample is under the effect of quantitative sheath fluid stream, cell is that single arrangement is also one by one through the aperture of detection zone A2 middle and lower reaches, the two ends up and down of detection zone A2 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, red blood cell and enumeration of thrombocytes mode are: determine cell volume by the measurement of impulse magnitude, obtain Cytometric result by the number of recording impulse, due to blood platelet (blood platelet, PLT) obvious difference is had with erythrocyte volume, difference according to blood platelet and erythrocyte volume sets threshold value, pulse signal higher than threshold value is defined as red blood cell, pulse signal lower than threshold value is defined as blood platelet, thus carries out red blood cell and enumeration of thrombocytes and measure analysis by the number of produced electronic impulse and size, wherein, quantity of leucocyte impact is negligible.The present invention accelerates the speed of stream of cells by detection zone by adopting the mode of secondary sheath stream, improve and detect cell quantity, accurate chip structure and pressure equilibrium simultaneously controls, the diameter of stream of cells is made to be stabilized in the width be close with haemocyte, guarantee that cell accepts electrical impedance and detects under hydrodynamic action in single arrangement, ensure to carry out measuring one by one, accurately and rapidly to a large amount of cell, and shorten the time of pattern detection.
Sheath Flow Technique is specially: in testing process, cell occurs side by side or lateral flow district after testing, and the phenomenon such as cell backflow, turbulent flow or eddy current brings metrical error, adopt sheath Flow Technique, namely cell suspension sample is under the effect of swiftly flowing sheath fluid lateral compression effect, formed similar enter the compression flow form of sheath shape, ensure that the bag of sample cell at sheath fluid is by the stream of cells of the single arrangement of lower formation, passes through detection zone successively; Sheath Flow Technique can be applicable to two kinds of cell count principles: one is theory of electrical impedance, and cell count is carried out in the sensitizing range that sheath flows through detection zone; Another kind is laser counting principle, and cell liquid stream room is longer, crossing with laser vertical, and laser beam produces light scattering to after each cell irradiation flowed through, and utilizes this principle to carry out cell count; Floating landmark technique is: because various intercellular boundary, therefore can be called floating landmark technique left with cell actual size or move right; In normal specimen, red blood cell differs comparatively large with volume of platelets, and generally red blood cell and hematoblastic boundary are scheduled 35fl, large is red blood cell, and little is blood platelet, what also to have with 30fl or 20fl be boundary; But large platelet may be had more than 35fl boundary under some pathologic condition, cause blood platelet to leak meter and make Lower result; Otherwise, if erythrocyte volume (as hypoferric anemia or thalass emia) less than normal, then may be blood platelet by part microcyte error count, make platelet count higher; In order to result is accurate, calculating instrument utilizes computing machine between 5-35fl, find histogram minimum point, be decided to be red blood cell and hematoblastic boundary with this, histogrammic boundary mark can make corresponding adjustment automatically according to the change of cell mass, and counted numerical value can be made thus to tally with the actual situation.
In embodiments of the present invention, liquid storage tank sample introduction adopts Micropump, electrokinetic injection, positive pressure to drive the various ways such as sample introduction, Ngatively pressurized sampling or electric osmose sample introduction, blood cell analysis chip can be made by materials such as quartz, glass, monocrystalline silicon, high molecular polymerizations, such as polymetylmethacrylate, polydimethylsiloxane or polycarbonate etc.; By adding the modes such as suitable adjuvant in chip microchannel modifying inner surface, chip material modification or solution to alleviate or to avoid chip microchannel surface to Hemadsorption; Simultaneously, the present invention, except being applied to medical industry, being equally applicable to relate to atomic quantity in the diameter of measurement of species particle or liquid and carrying out quantitatively and the industry such as analysis qualitatively, such as, in physico-chemical analysis in pure water, measure the content of its impurity and bacterium; Or the measurement of the degree of purity of various industrial high purity liquid or demarcation etc.
Referring to Fig. 4, is the process flow diagram of the chip type blood cell analysis method of the embodiment of the present invention.The chip type blood cell analysis method of the embodiment of the present invention comprises:
Step 400: detect the leucocyte on leucocyte/hemoglobin analysis chip by anticoagulation, dilution, hemolytic agent, leucocyte stabilizing agent and sheath fluid etc., and determine each subgroup character of leucocyte;
In step 400, white blood cell detection process is: by anticoagulation, dilution and hemolytic agent difference sample introduction are to liquid storage tank S1, R1, R2, whole blood suitably dilutes, and because of hemolytic agent osmotic pressure official post erythrocytolysis destruction, the effect with hemolytic agent in leucocyte stabilizing agent is added by liquid storage tank R5, make leukocyte surface, endochylema and cell volume return to ortho states substantially, sheath fluid sample introduction is to R6 liquid storage tank, blood sample is under the effect of sheath fluid stream, leucocyte is in line and one by one by detection zone A1, carry out one by one to the leucocyte of process, simultaneously, triple VCS detects, and by determination data as calculated machine process to determine each subgroup character of leucocyte.Because different classes of cell can present obvious difference in information characteristic such as volume, surface characteristics, inner structures, these information characteristic are defined in the three-dimensional scatter diagram formed for three-dimensional coordinate with VCS, five class leucocytes can form specific cell mass in three dimensions, accounting for the number percent of total white blood cells by calculating certain group of cell quantities, five leukocyte differential count results can be obtained; In embodiments of the present invention, to leukocytic detection except five leukocyte differential counts, can also measure and analyze the parameters such as the volume of neutrophil leucocyte, caryoplasm ratio, cell granulations characteristic, for understanding the difference of bacterium perception disease and Other diseases, also there is granulophilocyte analytic function and t lymphocyte subset cluster analysis function; Wherein, VCS is detected as: V-low frequency wave, analysis of cells volume; C-high frequency waves, analysis of cells caryogram; S-laser, the particle characteristics of analysis of cells; VCS detects and namely uses V, C, S tri-kinds of probes, in a certain site of detection zone, to the single-row leucocyte passed through carry out one by one, simultaneously, triple detections and three dimensional analysis, to determine its subgroup character; Specifically seeing also Fig. 5, is the process flow diagram of the white blood cell detection method of the embodiment of the present invention.The white blood cell detection method of the embodiment of the present invention comprises the following steps:
Step 401: adopt impedance bioelectrical measurement leucocyte volume;
In step 401, leucocyte volume measuring method is: certain low-frequency current is by two electrodes of sensor, because cell is non-conducting, when cell is by two electrodes, inter-electrode impedance increases instantaneously, the electric pulse that formation amplitude is directly proportional to cell volume, measures the volume of cell according to the size of pulse, the number according to pulse carries out cell count; Through to various cell produce the electronic selection of impulse magnitude, can the different types of cell of incomplete differentiation, the comparatively significant lymphocyte of effective differentiation volume difference in size and monocyte, but because pulse height that the is dissimilar but cell that volume is identical generation is identical, therefore only can not distinguish completely by cell volume, and then utilize high-frequency electrical inducing defecation by enema and suppository and laser light scattering analysis leucocyte inner structure.
Step 402: by high frequency conductance technology reacting cells internal structural information;
In step 402, concrete reactive mode is: although cell membrane can not make low-frequency current pass through, by high-frequency current; Electrode adds radio frequency (high frequency) generator, cell membrane has conduction to high-frequency current, when electric current forms alternating electromagnetic field around by during cell, form electromagnetic wave, because the conduction parameter of difference to high-frequency current between the ratio of nucleus, slurry, the inner structure such as particle and nucleocytoplasmic ratio is distinguished to some extent, the electromagnetic field formed around it is not identical yet, can reflect nuclear internal structural information by cell to the conduction parameter of high-frequency current and the variable quantity of electromagnetic field; And leaflet core and non-segmented cell can be distinguished, for distinguishing the cell colony that volume is close and inner structure is different, as lymphocyte and basophilic granulocyte.
Step 403: by laser scattering technology analysis of cells particle information;
In step 403, concrete analysis mode is: scanned each cell by one-wavelength laser, analysis of cells film surface characteristics and inner structure, detect grain pattern and density case in intracellular nucleic leaflet situation and endochylema, detect the light scattering characteristic of cell 10 ° of-70 ° of wide range, and distinguish the different cell colony of particle characteristics according to the light scattering characteristic of cell, the scattered light thinner than particle that such as intracellular granular is thick is stronger, distinguishes neutrophilia, acidophilia and the basophilla three kinds of cells in granulocyte whereby.
Step 500: the concentration adopting COLORIMETRY METHOD IN HEMOGLOBIN DETERMINATION;
In step 500, because red blood cell is discharged haemoglobin by dissolved destruction, haemoglobin and hemolytic agent react and form stable haemoglobin compound, haemoglobin compound enters haemoglobin test macro-colorimetric pool E, colorimetric under the specific wavelength of 530-550nm also measures its absorbance, in the change of absorbance and liquid, the content of haemoglobin is proportional, can record the concentration of haemoglobin thus;
Step 600: adopt sheath Flow Technique, electrical impedance technology or floating landmark technique to count respectively the red blood cell on red blood cell/plaque assay chip and blood platelet;
In step 600, red blood cell and platelet count process are: anticoagulation sample introduction is to liquid storage tank S2, dilution sample introduction is to liquid storage tank R5, quantitative whole blood sample dilutes by proper proportion through quantitative dilution, first sheath fluid sample introduction is to R6 liquid storage tank, second sheath fluid sample introduction is to R7 liquid storage tank, blood sample is under the effect of quantitative sheath fluid stream, cell is that single arrangement is also one by one through the aperture of detection zone A2 middle and lower reaches, the two ends up and down of detection zone A2 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, C8 to C13 is flow sensor, for making corresponding fluids in each liquid storage tank quantitative, red blood cell and enumeration of thrombocytes mode are: determine cell volume by the measurement of impulse magnitude, obtain Cytometric result by the number of recording impulse, due to blood platelet (bloodplatelet, PLT) obvious difference is had with erythrocyte volume, difference according to blood platelet and erythrocyte volume sets threshold value, pulse signal higher than threshold value is defined as red blood cell, pulse signal lower than threshold value is defined as blood platelet, thus carries out red blood cell and enumeration of thrombocytes and measure analysis by the number of produced electronic impulse and size, wherein, quantity of leucocyte impact is negligible.
Technical scheme tool of the present invention has the following advantages or beneficial effect: the chip type blood cell analyzer of the embodiment of the present invention and the somatotype of method by utilizing blood cell analysis chip to realize haemocyte, 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 particularly useful for the use of Site Detection, emergency analysis, domestic. applications and different medical unit.
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 (6)
1. a chip type blood cell analyzer, it is characterized in that, comprise blood cell analysis chip, described blood cell analysis chip comprises leucocyte/hemoglobin analysis chip and red blood cell/plaque assay chip, described leucocyte/hemoglobin analysis chip adopts electrical impedance, high frequency conductance and laser light scattering detection technique to detect leucocyte, and described red blood cell/plaque assay chip adopts sheath Flow Technique, electrical impedance technology or floating landmark technique to count respectively red blood cell and blood platelet;
Described leucocyte/hemoglobin analysis chip is provided with liquid storage tank, waste liquid pool, detection zone, colorimetric pool and multiple flow sensor, described liquid storage tank comprises liquid storage tank S1, liquid storage tank R1, liquid storage tank R2, liquid storage tank R5 and liquid storage tank R6, described liquid storage tank S1 is for storing anticoagulation, described liquid storage tank R1 is for storing dilution, described liquid storage tank R2 is for storing hemolytic agent, described liquid storage tank R5 is for storing leucocyte stabilizing agent, described liquid storage tank R6 is for storing sheath fluid, described leucocyte/hemoglobin analysis chip is also provided with first fluid passage, second fluid passage, 3rd fluid passage, 4th fluid passage, 5th fluid passage and sense channel, described liquid storage tank S1 is communicated with described waste liquid pool with described sense channel by described first fluid passage, described liquid storage tank R1 is communicated with described waste liquid pool with described sense channel by described second fluid passage, described liquid storage tank R2 is communicated with described waste liquid pool with described sense channel by described 3rd fluid passage, described second fluid passage and described 3rd fluid passage and described sense channel cross at same position, described liquid storage tank R5 is communicated with described waste liquid pool with described sense channel by two articles of described 4th fluid passages, article two, described 4th fluid passage and described sense channel cross at same position, described liquid storage tank R6 is communicated with described waste liquid pool with described sense channel by two articles of described 5th fluid passages, article two, described 5th fluid passage and described sense channel cross at same position, described second fluid passage and described 3rd fluid passage are in the position that crosses of described sense channel, article two, described 4th fluid passage is in the position that crosses of described sense channel, article two, described 5th fluid passage is in the position that crosses of described sense channel, described detection zone and described colorimetric pool are successively set on sense channel, and described second fluid passage and described 3rd fluid passage in the position that crosses of described sense channel near described liquid storage tank R1, described colorimetric pool is near described waste liquid pool, described first fluid passage is located at respectively by multiple described flow sensor, second fluid passage, 3rd fluid passage, on 4th fluid passage and the 5th fluid passage, described waste liquid pool is for storing the blood sample through detecting, described detection zone is used for detecting the leucocyte of process, determine each subgroup character of leucocyte, described colorimetric pool is for detecting the concentration of haemoglobin in blood sample, described flow sensor is used for making corresponding fluids in liquid storage tank quantitative.
2. chip type blood cell analyzer according to claim 1, is characterized in that, described leucocyte/hemoglobin analysis chip adopts described impedance bioelectrical measurement leucocyte volume; Adopt described high frequency conductance technology reacting cells internal structural information; Adopt described laser scattering technology analysis of cells particle information.
3. chip type blood cell analyzer according to claim 2, is characterized in that, described leucocyte/hemoglobin analysis chip adopts the concentration of colorimetric determination haemoglobin; The detection mode of the concentration of described detection haemoglobin is: colorimetric under the wavelength of 530-550nm also measures the absorbance of blood sample.
4. chip type blood cell analyzer according to claim 1, it is characterized in that, described red blood cell/plaque assay chip is provided with liquid storage tank, waste liquid pool, detection zone and flow sensor, described liquid storage tank is for storing detection reagent, described waste liquid pool is for storing the blood sample through detecting, the two ends up and down of described detection zone apply steady current, through electrical impedance technology detect make through cell produce electronic impulse, height according to pulse counts respectively to red blood cell and blood platelet, and described flow sensor is used for making corresponding fluids in liquid storage tank quantitative; Wherein, described red blood cell and enumeration of thrombocytes mode are: determine cell volume by the measurement of impulse magnitude, obtain Cytometric result by the number of recording impulse; Difference according to blood platelet and erythrocyte volume sets threshold value, pulse signal higher than threshold value is defined as red blood cell, pulse signal lower than threshold value is defined as blood platelet, passes through the number of produced electronic impulse and size and carry out red blood cell and enumeration of thrombocytes and measure analysis.
5. the chip type blood cell analyzer according to claim 1 or 4, is characterized in that, described liquid storage tank input mode comprises Micropump, electrokinetic injection, positive pressure driving sample introduction, Ngatively pressurized sampling or electric osmose sample introduction.
6. the chip type blood cell analyzer according to any one of Claims 1-4, is characterized in that, described blood cell analysis chip material comprises quartz, glass, monocrystalline silicon or macromolecule polymeric material.
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| EP3133395B1 (en) | 2014-04-17 | 2019-12-25 | Sony Corporation | Blood condition analysis device, blood condition analysis system, blood condition analysis method, and blood condition analysis program for enabling computer to perform said method |
| JP6442858B2 (en) | 2014-04-17 | 2018-12-26 | ソニー株式会社 | Blood state analysis apparatus, blood state analysis system, blood state analysis method, and blood state analysis program for causing a computer to realize the method |
| CN105728069B (en) * | 2016-01-30 | 2021-01-19 | 深圳市安测健康信息技术有限公司 | Multi-channel micro-fluidic chip for rapidly self-checking blood |
| CN106901706A (en) * | 2017-03-10 | 2017-06-30 | 安徽通灵仿生科技有限公司 | A kind of children gloves formula remote diagnosis system |
| CN107213930B (en) * | 2017-07-27 | 2022-12-20 | 深圳中科芯海智能科技有限公司 | Microfluidic chip for particle analysis and particle analysis method |
| 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 |
| CN108627448A (en) * | 2018-06-05 | 2018-10-09 | 江苏卓微生物科技有限公司 | The method of counting micro particles |
| CN115931667B (en) * | 2022-07-26 | 2024-01-05 | 中国石油大学(华东) | Method for evaluating permeability of hydrate sediment sample based on complex conductivity parameter |
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| CN103170377B (en) * | 2011-12-20 | 2015-02-18 | 中国科学院深圳先进技术研究院 | Hemocyte analysis chip and system for using chip thereof |
| CN103175950B (en) * | 2011-12-20 | 2015-04-22 | 中国科学院深圳先进技术研究院 | Hemocyte analysis chip and system for using chip thereof |
| CN102600918B (en) * | 2012-03-09 | 2014-06-18 | 深圳联合医学科技有限公司 | Microfluidic chip for blood analysis and method for using microfluidic chip |
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