CN1166951C - Woundless blood cell parameter measuring method - Google Patents

Woundless blood cell parameter measuring method Download PDF

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CN1166951C
CN1166951C CNB021336725A CN02133672A CN1166951C CN 1166951 C CN1166951 C CN 1166951C CN B021336725 A CNB021336725 A CN B021336725A CN 02133672 A CN02133672 A CN 02133672A CN 1166951 C CN1166951 C CN 1166951C
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image
blood
blood cell
haemocyte
volume
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CN1399131A (en
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彭黎明
吴大可
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彭黎明
吴大可
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Abstract

The present invention relates to a quantitative woundless blood cell parameter measuring method. Optical cohere tomography (OCT) is utilized, a light source with wideband laser is used for irradiating different focal depth parts of blood vessels in the tissues of a human body for zone focusing and image fusion reconstruction so as to obtain the OCT images of the three dimensional structures of various blood cells in the blood vessels having different dislocation depth, the obtained OCT images are analyzed and calculated, and the characteristic three dimensional structure information of complete blood cells comprising red blood cells, white cells and platelets and corresponding blood cell parameters are obtained. Therefore, blood collection through puncture for measuring the complete blood cells is avoided, and diseases can be diagnosed and treated.

Description

Woundless blood cell parameter measuring method

Technical field

The invention relates to the method for haemocyte being carried out quantitative measurement in non-invasive mode.In more detail, be by with optical coherence tomography (optical coherence tomography, OCT) technology method that the haemocyte that flows in the blood vessel is in vivo carried out quantitative measurement.

Background technology

Can obtain haemocyte component content by complete blood count (CBC) and comprise red blood cell (RBC), haemoglobin (Hb), leucocyte (WBC), blood platelet (PLT), hematocrit value (Hct), mean corpuscular volume (MCV), mean corpusular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC) (MCHC), red cell distribution width (RDW), average platelet volume (MPV), thrombocytocrit (Pct), the volume of platelets dispersion of distribution (PDW), neutrophil cell (NEUT) content and percent, lymphocyte (LYM) content and percent, monocyte (MONO) content and percent, eosinophil (EOS) content and percent, parameters such as basophilic granulocyte (BASO) content and percent are one of inspection items the most frequent in the clinical trial deagnostic test, and its detection comprises anaemia to clinical disease, infect, lose blood, leukopenia, decrease of platelet, the diagnosis of acute and chronic diseases etc. and treatment and important.

Gather the inevitable many unfavorable factors of traditional approach that vein or tip blood carry out CBC mensuration for a long time clinically by puncturing by solving, at present, existing people proposes some and is used for carrying out non-invasive analysers that above-mentioned some projects detect through skin.For example, Japanese HEI-3-71135 patent report the non-invasive analyser measured of a kind of haemoglobin (Hb), it is according to measuring Hb concentration owing to project the intensity variation that pulsation caused of the light of the multi-wavelength on the live body.Similarly, the U.S. the 5th, 372, No. 136 patent reports by utilize pulsation and similarly method measure the system and method for the hematocrit value in the blood.But these are used for determining technology of absolute value problem relevant with precision that invariably accompany, because blood volume can not be determined.And, can estimate the human body parts that measured value may tied up along with sensor and change, thereby cause that Hb result's is repeated relatively poor.

1991, the U.S. the 4th, 998,533 patent reports a kind ofly carry out the non-invasive analyser that RBC and WBC measure according to the blood-stream image in the capillary, this device has adopted graphical analysis and reflecting spectrograph measuring the parameter such as the cell size of individual cells, but can not carry out quantitative test to above-mentioned project.Simultaneously, measure and only can in capillary, carry out, and need large-scale structure.1998, publication number was that the Chinese patent literature of CN1200657A has also been reported with reflected image and carried out WBC, Hb, RBC, Hct and PLT in the non-invasive analysis blood.This technology utilizes the catoptric imaging method to the blood imaging, to produce a primary reflection image from reflecting less than the degree of depth of scattering length repeatedly, then this original image is proofreaied and correct, to form a reflected image of proofreading and correct, from the reflected image of proofreading and correct, intercept a scene again, forming an analysis image, and it is analyzed, comprise WBC, Hb, RBC, Hct and PLT to obtain the part blood parameters.This technology need be used crossed polarizers, and complex structure, and can not observe and detect the form and the three-dimensional structure of various haemocytes is just measured according to the difference of the reflected image of different haemocytes.Therefore, the accuracy of testing result and authenticity are not good enough.

For this reason, at medical domain, particularly in the clinical detection field, need a kind of based on the analytical approach that non-invasive CBC quantitative measurement is carried out in the three-dimensional structure image of different haemocytes and the analysis of blood transmission image, that is: provide in vivo various different haemocytes to comprise RBC, WBC, PLT form in noninvasive mode, and its form size and volume are provided the high resolving power demonstration and analyze; Can calculate the content of different haemocyte components and correlation parameter simultaneously according to the quantization characteristic of various haemocytes.This non-invasive blood cell analysis method should analyze quantitatively and be clinical detection required various live body haemocytes and correlation parameter, comprise 19 of the content of the content of the content of the content of the content of RBC, WBC, PLT, Hct, MCV, RDW, MPV, Pct, PDW, NEUT and percent, LYM and percent, MONO and percent, EOS and percent, BASO and percent etc., and the Hb content of reporting in conjunction with above-mentioned document and MCH, the MCHC etc. that can obtain thus totally 22 blood cell parameters.

Summary of the invention

According to above-mentioned situation, the present invention will provide a kind of method that adopts non-invasive mode can carry out quantitative measurement to multiple blood cell parameter.Specifically, be a kind of optical coherence tomography (optical coherencetomography that utilizes, being OCT) technology carries out non-invasive mensuration to the haemocyte that flows in the blood vessel in vivo, comprises the method for three-dimensional structure, form and the quantity of RBC, WBC, PLT being carried out quantitative test.

The OCT technology is based on the imaging art of optical fiber, its ultimate principle is similar to ultrasonic echo art (ultrasonography), utilizes the difference that the domain reflection-based mensuration art of the interference of light (optical coherence-domain reflectrometry) detection of reflected postpones between the light time or the difference of reflected light optical path.It is when organizing by wide band LASER Light Source irradiating biological, because various tissues or cell just can carry out imaging to various histiocytic microstructures to the difference of relevant (short-temporal coherence) degree in short-term of the broadband laser light source of incident.The performance parameter of decision OCT has the centre wavelength of longitudinal frame, lateral resolution, dynamic range, reference mirror translational speed and light source.Wherein, longitudinal frame is by the decision of the coherent length (coherence length) of light, wide being inversely proportional to of ripple (the Lc=λ of itself and light source 0 2/ Δ λ, wherein Lc is a longitudinal frame, λ 0 2Be lambda1-wavelength, Δ λ is the half-breadth height).At biomedical sector, OCT has been used to the microstructure imaging to the transparent or opaque biological tissue of live body original position at present.At first, OCT mainly is by the imaging to ocular tissue, is used for the pathology of retina and eye conjunctival tissue is checked.Since OCT vertically with the improving of lateral resolution, OCT has now begun also to be used for to body skin and multiple tube chamber system increasingly extensively, comprises that as the lesion tissue of alimentary canal, urogenital tract inflammation and tumour carry out histopathological examination.

Assay method of the present invention is on the basis of OCT technology, and the method that has adopted zone focusing and image co-registration to handle simultaneously again the steps include:

1) is suitable for haemocyte in the intravital blood vessel is carried out the broadband laser light source of image detection with wavelength, 5mm to biological tissue postpones in the degree of depth with the optical interference method, carry out zone focusing and obtain the OCT image of several separations at different depths of focus place with segmented mode, then each separate image that is obtained is merged and rebuild, thereby produce the coherence tomography image of the three-dimensional structure of haemocyte in the different tomography degree of depth blood vessels

2) resulting coherence tomography image of last step is carried out image signal's collection and analysis, comprise: from the image that has obtained, extract the characteristic three-dimensional structure picture signal that needs detect haemocyte, quantize the architectural feature of its image then, calculate corresponding blood cell parameter result according to this quantization characteristic again, and result of calculation is exported.

Carry out the quantity of zone focusing and the separate image that obtains respectively for said in the said method at different depths of focus place with segmented mode, test shows, chooses 40~50 separate images generally speaking and can obtain satisfied effect.

Further, when adopting said method that biological tissue is carried out zone focusing and obtains several separate images respectively with segmented mode at different depths of focus place, as with detected biological tissue position between light source and image capture part by relative fixed, can make the blood cell image in the intravital blood vessel that is collected even more ideal undoubtedly.

Adopt said method of the present invention, after OCT imaging and zone focusing and image co-registration are handled, can obtain different haemocytes in the biological tissue blood vessel---wherein should comprise the three-dimensional structure image of at least a or whole haemocytes in red blood cell, leucocyte and the blood platelet at least.

By said method, after gathering, to handle and analyze by the red blood cell in the intravital blood vessel (RBC), leucocyte (WBC) and blood platelet blood cell image data such as (PLT), its main contents and scope can comprise:

A. in to graphical analysis, make the RBC counting, to determine the RBC quantity of unit volume blood;

B. the RBC volume of measuring unit's volume blood in analysis image is to determine hematocrit value (Hct);

C. by graphical analysis, determine mean corpuscular volume (MCV) to RBC;

D. to RBC quantity and measure analysis the time, determine red cell distribution width (RDW);

E. in to graphical analysis, make the PLT counting, to determine the PLT quantity in the unit volume blood;

F. to PLT quantity and measure analysis the time, determine the volume of platelets dispersion of distribution (PDW);

G. by graphical analysis, determine average platelet volume (MPV) to PLT;

H. the PLT volume of measuring unit's volume blood in analysis image is to determine thrombocytocrit (Pct);

I. in to graphical analysis, make the WBC counting, to determine the WBC quantity in the unit volume blood;

J. in WBC quantity and graphical analysis, determine neutrophil cell (NEUT) content and percent;

K. in the quantity and graphical analysis of WBC, determine to comprise the leukocytic content of other types and the percent of lymphocyte (LYM), monocyte (MONO), eosinophil (EOS) and basophilic granulocyte (BASO).

The detected object that said determination method of the present invention is suitable for can comprise the live bodies of various animals, particularly comprises people's various mammals; Detected biological tissue part also should be this tissue part when being organized in the live body, for example finger or ear-lobe.On the other hand, the blood vessel that is comprised in the surveyed area is not particularly limited, but, in order to obtain the good result of reappearance, the preferably capillary of as close as possible skin or little artery and vein.In addition, the haemocyte information that is obtained by capillary or little artery and vein can convert the haemocyte information of big-and-middle artery and vein to.

For this reason, when adopting assay method of the present invention in employed instrument or the equipment, at least should include: a light source that is used to shine intravital blood and tissue, a harvester that is used to obtain the optical coherence tomography of haemocyte three-dimensional structures such as RBC, WBC, PLT, one is used to analyze the analysis part that institute's pickup image comprises the OCT image, wherein analysis part is analyzed the OCT image of haemocytes such as the RBC that gathers, WBC, PLT, thereby calculates haemocyte component content and output result of calculation.

Adopt OCT technology and zone focusing and image co-registration mode when obtaining the coherence tomography image of required haemocyte three-dimensional structure in the different tomography degree of depth blood vessels in the inventive method, employed light source generally can be LASER Light Source, wherein is suitable in vivo the broadband nanosecond laser source wavelength scope of RBC, WBC and PLT image detection and generally can selects in the scope of 600~1300 nanometers (nm).

The coherence tomography image that adopts haemocyte three-dimensional structure in the blood vessel that zone focusing and image co-registration mode obtained is analyzed and handled, should comprise at least being used to gather the extraction part of the optical coherence tomography of haemocytes such as RBC, WBC and PLT as its three-dimensional structure image; Be used to quantize the OCT characteristics of image of RBC, WBC and haemocytes such as PLT to calculate the calculating section of relevant blood component content; Be used for according to RBC, WBC and the OCT characteristics of image calculating of haemocytes such as PLT and the calculating section of the relevant blood component of haemocyte content such as RBC, WBC and PLT; And the efferent that is used to export result of calculation grades.

In order correctly to discern the different types of haemocyte of explanation, in analysis part, also should have the automatic identification equipment of the different haemocyte OCT images of identification according to the various haemocyte OCT images that absorbed.This device can carry out the comparison of cell size, nuclear/slurry form, nuclear/slurry ratio, chromatin Structure and particle etc. to haemocyte OCT image and the given normal plasma cell OCT reference picture that the test section obtains, and differentiates the OCT image of different haemocytes then according to comparative result.These work are easy to finish by computing machine, needed just according to the needs that detect and/or differentiate, the normal data relevant with detecting content imported computing machine in advance, the reference data standard of when analyzing and/or relatively differentiate, using as computing machine.

In above-mentioned analysis part,, utilize the pick-up unit of blood cell image the haemocyte that detects can be counted by variety classes by the OCT camera head of haemocyte is gathered the OCT image of haemocyte with period demand to same surveyed area; Simultaneously, can calculate the displacement of blood cell image, go out the translational speed of haemocyte, and then calculate the blood cell count of unit volume according to the translational speed of haemocyte according to its displacement and Image Acquisition computation of Period.

Image capture partly comprises the 3-D view of haemocytes such as picked-up RBC, WBC, PLT, the image capture part can be made of an optical system respectively, for example, it can comprise lens and image-pickup device, as the charge-coupled device (CCD) or the other forms of video camera of at present existing more use, haemocyte three-dimensional structure coherence tomography image in the picked-up blood vessel.The optical system of image capture part can be made of as adopting the equipment of being produced by COSMICAR company such as BD1214D TV lens.

As above-mentioned, assay method of the present invention adopts the broadband laser light source with the optical interference method the different depths of focus of biological tissue place to be carried out zone focusing and image is merged reconstruction, to obtain the coherence tomography image of various haemocyte three-dimensional structures in the different tomography degree of depth blood vessels, again two parts are analyzed and calculated to resulting optical coherence tomography image and form.By resulting optical coherence tomography image is analyzed, can obtain the characteristic three-dimensional structure information and the quantization characteristic of required detection haemocyte, calculate the parameter result of corresponding haemocyte.After image acquiring device carries out image acquisition to the same surveyed area with laser radiation, analysis part uses in several images of being gathered to form benchmark image earlier, calculate the poor of each Pixel Information to one in several images that obtained with benchmark image then, and form with the difference that calculates subtraction image as each Pixel Information.The result just can make that the background of image is eliminated in subtraction image, and the image that has only haemocyte is for observing and counting.

Test findings shows, non-invasive full blood cell analysis method of the present invention can above-mentioned 19 the clinical required blood cell parameters of quantitative measurement, for example can comprise content and the percent of the content of the content of the content of the content of RBC, WBC, PLT, Hct, MCV, RDW, MPV, Pct, PDW, NEUT and percent, LYM and percent, MONO and percent, EOS and percent, BASO.On this basis,, other relevant erythrocyte parameters be can also further obtain, mean corpusular hemoglobin (MCH) and mean corpuscular hemoglobin concentration (MCHC) (MCHC) comprised in conjunction with the Hb Determination on content data that obtain in the aforementioned documents mode.Wherein, MCH and MCHC can obtain by following computing formula:

MCH=Hb/RBC;

MCHC=Hb/Hct。

Like this, the whole blood cell analysis parameter can reach 22, makes resulting blood cell parameter comprehensive more, reasonable and practical.

Test findings shows, the non-invasive complete blood cell detection method that the present invention is above-mentioned is utilized advanced OCT optical coherence imaging technique, the resolution height, and clear picture can present imaging clearly to different cells, tissue simultaneously.Adopt the inventive method to detecting the picture depth of biological tissue, promptly longitudinal frame can be 3~5mm, and simultaneously, lateral resolution can be 10~15 μ m, and dynamic range is 150~200dB, and the degree of depth that sees through image of detection blood is not limit in tissue.By utilizing the optical coherence tomography feature of haemocytes such as RBC, WBC, PLT, promptly can determine the correlation parameter of RBC, WBC, PLT, Hct and other haemocytes of unit volume blood quantitatively.

In addition, advantage of the present invention also can show:

1. do not have wound, the wound of promptly having avoided puncture to cause has been cancelled by what blood was gathered in puncture the wound technology has been arranged.Like this, just eliminated from patients such as neonate, children, old age and burns and gathered blood for the misery of coming and difficult by puncture on one's body, simultaneously, got rid of the danger of patients with infectious diseases blood such as contact AIDS, hepatitis.

2. detect real-time, do not measure the CBC parameter with promptly can having wound fast and reach 22, and instant result can well be provided, therefore can be used as the other detection of bed for clinical use.

3. save cost, the needed detectable of wound technology is arranged by having saved tradition, and required sample transmission, processing and separating when detecting, thereby the total cost that detects saved.

4. testing result is accurate, accurate, utilizes optical coherence tomography to provide precondition for accuracy and the precision that improves CBC result's detection.

But 5. showed cell image can manifest the true three-dimension three-dimensional structure diagram of different haemocytes, in real time for analysis.

6. easy and simple to handle, need not the technical skill training.

7. purposes is wide, can be suitable for often monitoring the patient of CBC such as the patients such as neonate, children, old age and burn of tumour, hyperthyroidism, anaemia and blood sampling difficulty, also can be used for rescuing fast in extensive disaster and wartime.

According to foregoing, under the prerequisite that does not break away from the above-mentioned basic fundamental thought of the present invention,, can also make modification, replacement or the change of various ways easily to these contents according to the ordinary skill knowledge and the customary means of this area.

Below in conjunction with embodiment, foregoing of the present invention is described in further detail again by the accompanying drawing illustrated embodiment.But this should be interpreted as that the scope of the above-mentioned theme of the present invention only limits to following example.All technology that realizes based on foregoing of the present invention all belong to scope of the present invention.

Description of drawings

Fig. 1 is a kind of mode of operation synoptic diagram of assay method of the present invention,

Fig. 2 is the cross-sectional schematic of test section internal state among reflection Fig. 1.

Fig. 3 is the inventive method operating process synoptic diagram.

Fig. 4 is a kind of analyzing and processing schematic flow sheet to the optical coherence tomography image that is obtained among Fig. 3.

Fig. 5 is the synoptic diagram of reflection assay method analyzing and processing principle of the present invention.

Embodiment

As shown in Figure 1 be a kind of mode of operation of assay method of the present invention, the mensuration process can be divided into test section 1 and 2 two parts of carrying out continuously of analyzing and processing part.In test section 1, will be used to detect the live body part---finger 4 as shown in FIG. places the respective cavities 9 of the device that can be fixed, as shown in Figure 2.Be used for broadband nanosecond LASER Light Source 5 that finger tissue is shone and the scanning mirror 6 that is used for the blood vessel of finger tissue is sought the location by being arranged in the one side, and the image-pickup device 8 that has lens 7 on the other side, formed the image acquisition part, in order to obtain the OCT image of various haemocytes in the intravital blood vessel.Wherein used LASER Light Source for example can adopt the broadband nanosecond laser instrument of wavelength coverage between 600~1300 nanometers of Hamamatsu Photonics company.Image-pickup device 8 can be made up of two CCD, is used for the collection of haemocyte OCT image.When finger inserted in the cavity of opening 9, the capillary of light source 5 and blood vessel location scanning mirror 6 irradiation fingers 4 made the OCT image that different haemocytes produce in the blood vessel be absorbed by image-pickup device 8 scioptics 7.Light source 5 in the test section 1, seek the scanning mirror 6 and the image-pickup device 8 of location, can link to each other with analysis part 2 by cable 3 to organizing medium vessels.Analysis part 2 can be made up of personal computer.

OCT imaging analysis process to haemocytes such as RBC, WBC, PLT is: under the irradiation of broadband Bo microsecond LASER Light Source 5, the tissue blood vessel at 6 pairs of positions such as finger of scanning lens carries out horizontal and vertical scanning, obtains the OCT image of different haemocytes in the tissue blood vessel.At this moment, also can adjust and handle aberration by the reference mirror systems such as reference mirror, condenser lens and prism that comprise in the OCT device again, to obtain high-quality and distinct image.Like this, the OCT image of different haemocytes is gathered by CCD capturing apparatus 8, and picture signal enters next step analysis part 2 after amplification and opto-electronic conversion.In analysis part 2, to obtain that various haemocyte OCT image informations in the intravital blood vessel that comprises RBC, WBC, PLT etc. are gathered, image to having extracted, the feature that comprises the OCT image that RBC, WBC, PLT produce is carried out quantification treatment, is calculated various blood cell parameters according to the digitalized signature that quantizes gained, and for example 19 of aforesaid haemocyte parameters are calculated and result of calculation is exported through equipment such as CRT or printers.

Fig. 3 is the operating process signal that the non-invasive determination of blood cell method of the present invention detects haemocyte.As shown in Figure 3, in test section 1,, seek subcutaneous capillary (step C2), and set best analysis position (step C3) with focus lens system through the irradiation (step C1) of broadband laser device; Then, with image acquiring device CCD the same surveyed area of setting is carried out the OCT imaging and gathers (step C4) endovascular haemocyte.In analysis part 2, the haemocyte OCT image of being gathered is extracted and analyzes, comprise form, nuclear/slurry ratio, STRUCTURE OF CHROMATIN and the intracellular particle (step C5) of cell size, nuclear and slurry; Then, with in the computer the various haemocyte above-mentioned parameters of normal person stored of input in advance compare analysis (step C6), to determine the kind and the intensity of anomaly (step C7) of various haemocytes, count the content (step C8) of various haemocytes, and calculate the volume (step C9) of various haemocytes.

Fig. 4 is the flow process signal to a kind of analyzing and processing of the optical coherence tomography image that is obtained among Fig. 3.As shown in Figure 4, image-pickup device CCD is in the irradiation through broadband nanosecond laser instrument, after condenser lens is sought blood vessel and determined the optimized analysis position, begin to gather the OCT image of various haemocytes in the blood vessel, and this blood cell image gatherer process is continuous.Wherein, gathering the OCT image of haemocyte in the blood vessel first time is A (0)(step S1), then, the OCT image of gathering second haemocyte is A (1)(step S2).Then, measure image A (1)And A (0)Each pixel data poor forms with this and differs from subtraction image (step S3) as pixel data.This subtraction image is adjusted the processing (step S4) of contrast and the filtering processing (step S5) of elimination noise contribution.Threshold value with given different haemocytes is carried out binaryzation (step S6) with the pixel of such processed images, detects the haemocyte picture, the different haemocyte of identification from detect the haemocyte picture.For this reason, blood cell image and the variety classes that proposes normally compared (step S7) with reference to the OCT image of haemocyte.Then, check that degree (correlation) R of the two OCT picture registration is at certain value R 0Above person (step S8) is considered as corresponding haemocyte.As R at R 0Below, then can be judged as and be not and the corresponding haemocyte of haemocyte relatively, need carry out OCT graphical analysis (from step S2) again.Haemocyte as the mistake discerned in the haemocyte discerned and the previous subtraction image is same haemocyte, obtains distance, delta L between the two, and calculate translational speed V (step S9, S10, S11).If this haemocyte is new identification, then will count K and be made as K+1, and write down its position (step S12, S13).K is the blood count that is identified.The later step of analyzing with the defined of necessity (step S14) of OCT amount of images repeating step S2.Then, the mean value Va of use gained K and V by formula calculates the content of haemocyte (RBC, WBC, PLT), and its computing formula is:

Haemocyte (RBC, WBC, PLT)=AK/Va

Wherein A is a constant.Simultaneously, can use given function that the blood cell count in the capillary is converted into the blood cell count of corresponding big-and-middle artery and vein, this is because the blood cell count of unit volume blood is constant in big, medium vessels.

Fig. 5 has reflected the assay determination principle of assay method of the present invention, expression be the change procedure of haemocyte location during haemocyte detects in the methods of the invention.As shown in Figure 5, in a given time, as in 1/30 second time, some haemocytes move to position B from position A.Then, survey distance, and calculate the flow velocity V of this haemocyte from position A to B.Simultaneously, blood cell count is carried out above-mentioned K counting (step S12), can calculate actual blood cell count by these numerical value again.

Above-mentioned haemocyte can be meant RBC, WBC and PLT.After calculating the WBC sum, discern and count resulting various leucocyte again with by the step C5 among Fig. 3, C6, C7, the percent that comprises NEUT, LYM, MONO, EOS, BASO multiplies each other, and can obtain the quantity that the interior various leucocytes of unit volume comprise NEUT, LYM, MONO, EOS, BASO.

By step C5~C9 of Fig. 3 and step S2~S15 of Fig. 4, can determine simultaneously that RBC quantity and the hematocrit value in the unit volume is Hct.The formula that calculates Hct is:

Hct=A R·∑V R/Va

Wherein, A RBe constant, ∑ V RBe the summation of the erythrocyte volume of counting (f1).

Meanwhile, by step C5~C9 of Fig. 3 and step S2~S15 of Fig. 4, can also determine simultaneously that PLT quantity and the thrombocytocrit in the unit volume is Pct.The formula that calculates Pct is:

Pct=A P·∑V P/Va

Wherein, A PBe constant, ∑ V PIt is the summation of institute's platelet Counting volume (f1).

Step C5~C9 by Fig. 3 and step S2~S15 of Fig. 4 can count RBC quantity, but also the volume of measure R BC are Hct to the OCT graphical analysis of RBC, thereby determine MCV.Its formula that calculates MCV is:

MCV=Hct/RBC

Meanwhile, the step C5~C9 by Fig. 3 and step S2~S15 of Fig. 4 can count PLT quantity to the OCT graphical analysis of PLT, and the volume that also can measure PLT is Pct, thereby determine MPV.The formula that calculates MPV is:

MPV=Pct/PLT

By step C5~C9 of Fig. 3 and step S2~S15 of Fig. 4 the OCT image of RBC is analyzed, when counting RBC and its stereometry, its corresponding volume of RBC number according to measuring can calculate RDW, and its computing formula is:

RDW-SD=[∑(X R-meanX R) 2/(n-1)] 1/2

RDW-CV=SD/meanX R×100%

Wherein, SD is the standard deviation of RBC volume, X RBe the volume of single RBC, meanX RBe the average external volume of RBC, ∑ (X R-meanX R) 2Be the quadratic sum of the difference of the volume of all RBC and RBC average external volume, CV is the variation percent of RBC volume.

By step C5~C9 of Fig. 3 and step S2~S15 of Fig. 4 the OCT image of PLT is analyzed, when counting PLT and its stereometry, its corresponding volume of PLT number according to measuring can calculate PDW.Its computing formula is:

PDW-SD=[∑(X P-meanX P) 2/(n-1)] 1/2

PDW-CV=SD/meanX P×100%

Wherein, SD is the standard deviation of PLT volume, X RBe the volume of single PLT, meanX PBe the average external volume of PLT, ∑ (X P-meanX P) 2Be the quadratic sum of the difference of the volume of all PLT and PLT average external volume, CV is the variation percent of PLT volume.

In addition, measure resulting Hb by the non-invasive detection method that obtains by the aforementioned documents method, RBC and two parameters of Hct with above-mentioned can also further calculate other 2 erythrocyte parameters that comprise MCH and MCHC.Its computing formula is respectively:

MCH=Hb/RBC

MCHC=Hb/Hct

Thus, in this embodiment method of the present invention, reach the correlation computations of further carrying out, can finish quantitative measurement 19 parameters relevant with haemocyte by direct mensuration.Wherein, totally 4 of the parameters relevant with RBC comprise RBC, Hct, MCV and RDW; Totally 11 of the parameters relevant with WBC comprise WBC, NEUT (percent and content), LYM (percent and content), MONO (percent and content), EOS (percent and content) and BASO (percent and content); Totally 4 of the parameters relevant with PLT comprise PLT, Pct, MPV, PDW.In these parameters, 4 parameters relevant with RBC are to determine as follows:

RBC---directly measure,

Hct---directly measure,

MCV——Hct/RBC,

RDW-CV——SD/meanX R

11 parameters relevant with WBC are to determine as follows:

WBC---directly measure,

NEUT (%)---directly measure,

NEUT content---NEUT (%) * WBC,

LYM (%)---directly measure,

LYM content---LYM (%) * WBC,

MONO (%)---directly measure,

MONO content---MONO (%) * WBC,

EOS (%)---directly measure,

EOS content---EOS (%) * WBC,

BASO (%)---directly measure,

BASO content---BASO (%) * WBC;

4 parameters relevant with PLT are to determine as follows:

PLT---directly measure,

Pct---directly measure,

MPV——Pct/PLT,

PDW——SD/meanX P

By these parameters, will more help Clinics and Practices to disease with the complete blood cell of non-invasive mode quantitative measurement.

Claims (6)

1. Woundless blood cell parameter measuring method is characterized in that adopting the optical coherence tomography technology, the steps include:
1) with wavelength coverage is the broadband nanosecond LASER Light Source of 600~1300nm, 5mm to biological tissue postpones in the degree of depth with the optical coherence tomography method, carry out the optical coherence tomography image that the haemocyte of several separations is also obtained in zone focusing respectively at different depths of focus place with segmented mode, then each separate image that is obtained is merged and rebuild, thereby produce the coherence tomography image of the three-dimensional structure of haemocyte in the different tomography degree of depth blood vessels
2) the optical coherence tomography image that the last step was produced carries out image signal's collection and analysis, comprise: the characteristic three-dimensional structure picture signal of from the image that has obtained, extracting at least a haemocyte in red blood cell, leucocyte and the blood platelet, quantize its picture structure feature then, export again according to this quantization characteristic calculating other blood cell parameters correspondingly, and with result of calculation.
2. Woundless blood cell parameter measuring method as claimed in claim 1 is characterized in that carrying out zone focusing at different depths of focus place and obtaining 40~50 separate images respectively with segmented mode.
3. Woundless blood cell parameter measuring method as claimed in claim 1, it is characterized in that biological tissue is carried out zone focusing with segmented mode and when obtaining the image of several separations respectively at different depths of focus place, should detected biological tissue position light source and image capture partly between by relative fixed.
4. Woundless blood cell parameter measuring method as claimed in claim 1, it is characterized in that the optical coherence tomography image that focuses on and obtain respectively the haemocyte of several separations with segmented mode at different depths of focus place through the zone merges when rebuilding, use one in several images of being gathered to form benchmark image earlier, calculate the poor of each Pixel Information to one in several images that obtained with benchmark image then, and form with the difference that calculates subtraction image as each Pixel Information, image background in subtraction image is eliminated, and the image that only keeps haemocyte is for observing and counting.
5. Woundless blood cell parameter measuring method as claimed in claim 1 is characterized in that handling after being gathered by the blood cell image data of obtaining in the intravital blood vessel, and other blood cell parameters that calculate correspondingly according to this quantization characteristic comprise:
A. in to graphical analysis, make red blood cell count(RBC), to determine the erythrocyte number of unit volume blood;
B. the erythrocyte volume of measuring unit's volume blood in analysis image is to determine hematocrit value;
C. by to erythrocytic graphical analysis, determine mean corpuscular volume;
D. to erythrocyte number and measure analysis the time, determine red cell distribution width;
E. in to graphical analysis, make platelet count, to determine the platelet counts in the unit volume blood;
F. to platelet counts and measure analysis the time, determine the volume of platelets dispersion of distribution;
G. by to hematoblastic graphical analysis, determine the average platelet volume;
H. the volume of platelets of measuring unit's volume blood in analysis image is to determine thrombocytocrit;
I. in to graphical analysis, make white blood cell count(WBC), to determine the quantity of leucocyte in the unit volume blood;
J. in quantity of leucocyte and graphical analysis, determine neutrophil cell content and percent;
K. in leukocytic quantity and graphical analysis, determine to comprise the leukocytic content of other types and the percent of lymphocyte, monocyte, eosinophil and basophilic granulocyte.
6. Woundless blood cell parameter measuring method as claimed in claim 1, it is characterized in that when different tomography degree of depth blood vessels are carried out zone focusing and image co-registration mode, with haemocyte three-dimensional structure coherence tomography image in the charge-coupled device video camera picked-up blood vessel.
CNB021336725A 2002-08-28 2002-08-28 Woundless blood cell parameter measuring method CN1166951C (en)

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