CN106526154A - Measurement method of erythrocyte shear modulus, and measurement method of oxygen carrying capacity of blood - Google Patents
Measurement method of erythrocyte shear modulus, and measurement method of oxygen carrying capacity of blood Download PDFInfo
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- CN106526154A CN106526154A CN201611087788.XA CN201611087788A CN106526154A CN 106526154 A CN106526154 A CN 106526154A CN 201611087788 A CN201611087788 A CN 201611087788A CN 106526154 A CN106526154 A CN 106526154A
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/483—Physical analysis of biological material
- G01N33/487—Physical analysis of biological material of liquid biological material
- G01N33/49—Blood
Abstract
The invention provides a measurement method of an erythrocyte shear modulus, and a measurement method of the oxygen carrying capacity of blood. The measurement method of the erythrocyte shear modulus comprises the following steps: adding an incubation solution into a sample cell, wherein the incubation solution contains erythrocytes and dielectric microspheres, and the dielectric microspheres comprise first dielectric microspheres adhered to two opposite sides of the erythrocytes and second dielectric microspheres not adhered to the erythrocytes; capturing the second dielectric microspheres by using scanning tweezers, and calibrating the optical trap stiffness of the scanning optical tweezers; and capturing the first dielectric microspheres at any side of the erythrocytes by using the scanning tweezers to obtain a relationship curve between the relative elongation of the erythrocytes and the optical trapping force, and substituting the gradient of the curve into an erythrocyte shear modulus calculating expression in order to obtain the erythrocyte shear modulus. The measurement methods have the advantages of simplicity, few operation steps, great shortening of the measurement time, high measurement efficiency, and high measurement result accuracy. The oxygen carrying capacity of blood can be obtained through measuring the erythrocyte shear modulus of blood to be measured.
Description
Technical field
The present invention relates to biological technical field, and the measuring method and blood of more particularly to a kind of erythrocyte modulus of shearing are defeated
Oxygen ability measurement method.
Background technology
Erythrocyte major function is for biological tissue and conveyed oxygen, while research shows that erythrocyte has immune work(
Can, erythrocyte is played an important role in the normal operation of human physiological functions.In blood circulation, erythrocyte need to be by straight
Blood capillary of the footpath less than own dimensions, therefore deformability is significant for erythrocyte, becomes and examines biology, medical science
Examine the important indicator of blood oxygen carrying capacity.It is existing measurement erythrocyte membrane elasticity technology include laser diffractometry, viscosimetry with
And micropore screen method, these methods can only be used to the somatic elasticity of phenon, be capable of achieving the micro pipette method that individual cells are manipulated
Then because operation difficulty is greatly not with practicality.For the measurement of single celled elastic modelling quantity cannot be realized always.It is right at present
Optical tweezer technology is typically adopted in the measurement of individual cells film elasticity, but the operation of erythrocyte membrane elastic modelling quantity is measured using optical tweezer
Journey is relatively complicated.In the pretreatment of sample, for the modification of handle microsphere is constantly subjected to the favor of researchers.
Inventor's research finds that functionalized reagent modification handle microsphere is unknown to the impact for measuring, and microsphere modification consumption
When it is more, costly；In measurement operation, existing optical tweezer measuring method is the stress strain curve for measuring single erythrocyte, is needed
Erythrocyte is stretched with different power repeatedly, continually mobile example platform；Shortcoming above has all had a strong impact on erythrocyte membrane bullet
The measurement efficiency of property modulus.
The content of the invention
It is an object of the invention to provide a kind of measuring method of erythrocyte modulus of shearing, this measuring method measurement process behaviour
Make simple, measurement efficiency height and certainty of measurement is high.
Another object of the present invention is to a kind of measuring method of blood oxygen carrying capacity is provided, it is red thin in blood by measuring
The modulus of shearing of born of the same parents, obtains the oxygen carrying capacity of blood, and using value is big.
The present invention solves its technical problem and employs the following technical solutions to realize.
The present invention proposes a kind of measuring method of erythrocyte modulus of shearing, and which includes：
Addition step：Hatching solution is added into sample cell, in hatching solution, contains erythrocyte and electrolyte microsphere, electrolyte
Microsphere includes the first electrolyte microsphere for adhering to erythrocyte opposite sides and not micro- with the second electrolyte of red blood cell adhesion
Ball；
Optical Trap Stiffness demarcating steps：The second electrolyte microsphere is caught using scanning optical tweezer, the Optical Trap Stiffness to scanning optical tweezer
Demarcated；
Stretching step：Using the first electrolyte microsphere of any side of scanning optical tweezer capture erythrocyte, fitting obtains red thin
The relative elongation of born of the same parents and the relation curve of trapping stiffness, the slope of relation curve is k, wherein, relative elongation is erythrocyte quilt
The ratio of the diameter of extended length and erythrocyte after stretching, the slope k of relation curve are the ratio of relative elongation and trapping stiffness
Value；
Calculation procedure：The modulus of shearing of erythrocyte is calculated, computing formula is as follows：
Wherein, modulus of shearing of the H for erythrocyte, slopes of the k for relation curve, diameters of the d for erythrocyte, B are cut for distortion
Shear modulu, B=2 × 10-19Nm。
The present invention proposes a kind of blood oxygen carrying capacity measuring method, and which includes the measurement using above-mentioned erythrocyte modulus of shearing
Method measures the erythrocyte modulus of shearing of blood to be measured.
The invention has the beneficial effects as follows：
Using scanning optical tweezer technology, continuously elongated operation is carried out to erythrocyte, it is not necessary to repeatedly erythrocyte is stretched
Operation, frequent movable operating platform can simplify operating procedure, greatly shorten the time of experimental implementation.Demarcate scanning optical tweezer
Optical Trap Stiffness, then by the continuously elongated relative elongation for obtaining erythrocyte to erythrocyte and the relation curve of trapping stiffness, root
The modulus of shearing of erythrocyte can be calculated according to the slope k value of relation curve.The measuring method of the erythrocyte modulus of shearing is surveyed
Amount process is simple, easily operated, the efficiency high of measurement, and the high precision of measurement result.
Meanwhile, the modulus of shearing of blood rbc to be measured can simply, be quickly measured using above-mentioned measuring method, with health
The erythrocyte modulus of shearing of blood compares, and modulus of shearing is bigger, and the deformability of blood rbc to be measured is less, red
Cell is lower by the ability cured by oxygen therapy by blood capillary.Blood to be measured is obtained by the modulus of shearing of blood rbc to be measured
The size of liquid oxygen carrying capacity.
Description of the drawings
In order to be illustrated more clearly that the technical scheme of the embodiment of the present invention, below by to be used attached needed for embodiment
Figure is briefly described, it will be appreciated that the following drawings illustrate only certain embodiments of the present invention, thus be not construed as it is right
The restriction of scope, for those of ordinary skill in the art, on the premise of not paying creative work, can be with according to this
A little accompanying drawings obtain other related accompanying drawings.
Schematic diagrams of the Fig. 1 for the AOD scanning optical optical tweezers systems of the embodiment of the present invention；
Schematic diagrams of the Fig. 2 for the measuring method of the embodiment of the present invention, wherein Fig. 2 (a) are the schematic diagram before erythrocyte stretching；
Fig. 2 (b) is the schematic diagram after erythrocyte stretching；
Optical Trap Stiffness calibration process figures of the Fig. 3 for the embodiment of the present invention；
Relation curves of the Fig. 4 for the erythrocyte relative elongation and trapping stiffness of the embodiment of the present invention.
Icon：11- lasing light emitters；12- acousto-optic deflection devices；13- lens；14- dichroic mirrors；15- inverted microscopes；16-CMOS
Image pick-up device；17- ligh traps are moved；18- samples；19- illumination light；1- silicon dioxide microspheres；2- erythrocyte；3- optical tweezers；4- sample cells
Bottom.
Specific embodiment
To make purpose, technical scheme and the advantage of the embodiment of the present invention clearer, below will be in the embodiment of the present invention
Technical scheme be clearly and completely described.In embodiment, unreceipted actual conditions person, builds according to normal condition or manufacturer
The condition of view is carried out.Agents useful for same or the unreceipted production firm person of instrument, are the conventional product that can pass through that commercially available purchase is obtained
Product.
Below the measuring method of the erythrocyte modulus of shearing of the embodiment of the present invention is specifically described.
A kind of measuring method of erythrocyte modulus of shearing provided in an embodiment of the present invention.
Before experiment, the blood and electrolyte microsphere of healthy donor are gathered as laboratory sample, electricity using vacuum test tube
Medium microsphere is preferably the silicon dioxide microsphere of 2～4 μm of diameter.
Further, the silicon dioxide microsphere from a diameter of 4 μm, test result indicate that, the silicon dioxide under the diameter
Microsphere, the homogeneity of its particle diameter are higher, bigger with the sticking probability of erythrocyte.And when using warm-up movement analysis method demarcation ligh trap
During rigidity, the silicon dioxide microsphere of the particle diameter is captured with optical tweezer, the displacement that silicon dioxide microsphere deviates ligh trap center is less, ligh trap
The linear relationship of power and shift value is good, Optical Trap Stiffness high precision obtained by calibrating.
Using silicon dioxide microsphere as handle microsphere, silicon dioxide microsphere can be adhered to red with non-specific binding power
On cell, it is not necessary to modified using dressing agent, it is to avoid because dressing agent produces impact to result of the test.Simplify operation simultaneously
Step, saves operating cost.
Sample clean：
Respectively silicon dioxide microsphere, erythrocyte are cleaned.Specially：Silicon dioxide microsphere solution and blood sample point
Not Shi Yong phosphate buffer (PBS) dilution, silicon dioxide microsphere using be cleaned by ultrasonic 10～20min, 6000～8000rpm from
10～15min of the heart is collected, and is repeated 2～3 times；Erythrocyte is centrifuged 10～15min and is collected using agitation dispersion, 2000～3000rpm,
Repeat 2～3 times.
Incubation step：
The silicon dioxide microsphere for cleaning, erythrocyte are dispersed in PBS again, the concentration for adjusting erythrocyte is 1 × 105
～2 × 105Individual/μ l, and the mass concentration ratio of silicon dioxide microsphere and erythrocyte is adjusted for 2～4:1, will the two equal proportion mixing
Uniformly.30～60min of hatching is stood in 1～5 DEG C of environment to obtain hatching solution.Now, contain opposite sides in hatching solution equal
It is stained with the erythrocyte and the not silicon dioxide microsphere with red blood cell adhesion of silicon dioxide microsphere.The adhesion of erythrocyte opposite sides
Silicon dioxide microsphere is the first electrolyte microsphere, is not the second electrolyte with the silicon dioxide microsphere of red blood cell adhesion micro-
Ball.Further, the mass concentration ratio of silicon dioxide microsphere and erythrocyte is 2:1, under the mass concentration ratio, erythrocyte both sides pair
Claim the probability increase of two silicon dioxide microspheres of adhesion, it is ensured that preferably experiment condition, meet measurement demand.
In preferred embodiments of the present invention, incubation temperature is preferably 4 DEG C, and brooding time is preferably 40min.4 DEG C of environment
Under, it is ensured that the form of erythrocyte does not change, while under the brooding time, it is ensured that silicon dioxide microsphere and erythrocyte it is abundant
Adhesion.
Sample cell process step：
Dry serum protein layer is formed in sample cell bottom, preferably：Coverslip and microscope slide use 1～2% Ox blood serum
Protein solution soaks 15～30min and dries, and forms the sample cell that bottom is coated with a small amount of bovine serum albumin.In sample cell bottom shape
Into dry serum protein layer, the non-specific adhesion power of silicon dioxide microsphere and sample cell bottom can be reduced, it is red thin beneficial to controlling
Two kinds of silicon dioxide microsphere on born of the same parents adhere to the state of sample cell bottom, reduce the influence factor of measurement, it is ensured that measurement
Accuracy.
In preferred embodiments of the present invention, sample cell is dried in dustless environment, it is to avoid impurity contaminated samples pond.Sample
After product pond is dried, sample room is formed between coverslip and microscope slide using 50～100 μ m-thick plastic sheet pads.
Addition step：
The concentration of dilution hatching solution to erythrocyte is 1 × 103～2 × 103Individual/μ l, seal after being then added dropwise to sample room
Sample cell.Preferably, sample cell is closed using canada balsam, reduces impact of the air-flow to measuring, it is ensured that measurement
Accuracy and precision.
Optical Trap Stiffness demarcating steps：The nonadherent dioxy on erythrocyte is caught using the scanning optical tweezer under different luminous powers
SiClx microsphere, the Optical Trap Stiffness to scanning optical tweezer are demarcated.
When capturing to fine particle, fine particle is acted on optical tweezer by the power for pointing to ligh trap center, this
One force trapping is trapping stiffness.The displacement that trapping stiffness deviates ligh trap center to fine particle is directly proportional, i.e.,
F=-kxΔ x, (1)
In formula, F is trapping stiffness, kxFor Optical Trap Stiffness, Δ x is the displacement that fine particle deviates ligh trap center.
Therefore, the demarcation of Optical Trap Stiffness is the basis for measuring trapping stiffness.The scaling method of Optical Trap Stiffness includes power spectrum point
Analysis method, hydrodynamic methods, thermal motion analysis method, additional cycle force method etc..In preferred embodiments of the present invention, using heat fortune
Dynamic analytic process demarcates scanning Optical Trap Stiffness of the optical tweezer to silicon dioxide microsphere.
The calibration principle of thermal motion analysis method is：When capturing a fine particle with ligh trap, substantially can see from microscope
Observe, this fine particle does limited Brownian movement.The range of movement of fine particle is substantially all in the range of the simple harmonic quantity of ligh trap,
And Brownian movement of the target particles in ligh trap meets Boltzmann (Boltzmann) distributions.Temperature in sample cell is not very
Height, when the axial depth of ligh trap is not very shallow, fine particle Brownian movement in ligh trap will not be jumped out the simple harmonic quantity scope of ligh trap,
The potential field of ligh trap immediate vicinity is simple harmonic quantity potential field E (x), and the computing formula of E (x) is as follows：
The bias for deviateing ligh trap center of the fine particle that hypothesis ligh trap is captured is x, now the potential energy of fine particle
For E (x), the position distribution according to fine particle in ligh trap meets Boltzmann distributions, then herein at fine particle in this place
Probability p (x) it is as follows：
In formula, probability of the p (x) for fine particle, kBFor Boltzmann constant, a is normaliztion constant, and T is Kelvin.
By the position for testing silicon oxide pellets in big measurement ligh trap, the position distribution machine of silicon dioxide microsphere is counted
Rate, obtains E (x) values using formula (3), obtains afterwards scanning the Optical Trap Stiffness of optical tweezer.Using the scanning optical tweezer of different luminous powers
Silicon dioxide microparticle is captured, the relation curve of luminous power and Optical Trap Stiffness is obtained.As a result show, luminous power and ligh trap are firm
The linear relationship of degree is good, consistent with notional result, therefore the scaling method simple possible, and measurement result is accurate.
Stretching step：
Using the silicon dioxide microsphere of any side of scanning optical tweezer capture erythrocyte, continuously elongated erythrocyte, fitting are obtained
The relative elongation of erythrocyte and the relation curve of trapping stiffness, the slope of the relation curve is k.Wherein, the relative of erythrocyte is stretched
The ratio of the long diameter for measuring extended length and erythrocyte after being stretched for erythrocyte, the slope k of relation curve is specific elongation
The ratio of amount and trapping stiffness.
Calculation procedure：
The modulus of shearing of erythrocyte is calculated, computing formula is as follows：
Wherein, modulus of shearing of the H for erythrocyte, k are oblique with the relation curve of trapping stiffness for the relative elongation of erythrocyte
Rate, diameters of the d for erythrocyte, B are distortion modulus of shearing, B=2 × 10-19Nm。
In preferred embodiments of the present invention, optical tweezer is scanned using acousto-optic deflection device, i.e. it is firm that AOD scannings optical tweezer carries out ligh trap
The stretching of the labelling and erythrocyte of degree.The core component of scanning optical tweezer is beam deflector, scanning galvanometer, acousto-optic is usually used inclined
Turn the devices such as device (AOD), electro-optic deflector.
The embodiment of the present invention scans optical tweezer using AOD, and AOD scanning optical optical tweezers systems are as shown in Figure 1.What lasing light emitter 11 was launched swashs
Light beam modulates the propagation path of light by acousto-optic deflection device 2, through lens 13, dichroscope 14, puts down in the picture of inverted microscope 15
Quickly scan on face, ligh trap movement 17 is controlled by computer 20, laser beam is switched fast between several fixing points.Its
Optical Tweezers Array is formed in sample cell 18, microgranule can be captured or dynamic operation is carried out, by the scanning speed for controlling optical tweezer,
Captured particle can be made to move along the scanning track of light beam.Compared to the operation of monochromatic light tweezer and dual access test, scanned using AOD
Optical tweezer can greatly simplify operating procedure, to improving the efficiency of operation and the precision of operation.
The drawing process of erythrocyte is illustrated in figure 2, after hatching, two silicon dioxide microspheres 1 stick to 2 edge of erythrocyte
Diametric both sides, the silicon dioxide microsphere 1 of wherein 2 side of erythrocyte non-specifically stick to sample cell bottom 4, adopt
AOD scanning optical tweezers 3 capture the silicon dioxide microsphere 1 of opposite side, which are pulled away from from the bottom of sample cell, continuously elongated erythrocyte 2.
Preferably, along the continuously elongated erythrocyte in direction 2 of two 1 lines of silicon dioxide microsphere.It is further preferred that computer operation is red
The rate of extension of cell 2 is less than or equal to 0.02 μm/s, under this rate of extension, the adhesion of silicon dioxide microsphere 1 and sample cell
Power is negligible, and now, the pulling force size that erythrocyte 2 is subject to is equal with trapping stiffness so that the result of measurement is more accurate.
The continuously elongated process of each erythrocyte 2 is recorded and is preserved by image pick-up device.In drawing process, trapping stiffness,
The pulling force being applied on erythrocyte 2 constantly increases, and by the shift value Δ x at 1 center of silicon dioxide microsphere and ligh trap center, ties
Optical Trap Stiffness obtained by calibrating is closed, the real-time trapping stiffness of drawing process can be obtained.
By image analysis software analysis stretching video, the relative elongation for obtaining erythrocyte 2 is bent with the relation of trapping stiffness
Line, is fitted to the relation curve, obtains the slope k of relation curve, substitutes in formula (4), is calculated cutting for erythrocyte 2
Shear modulu.
In preferred embodiments of the present invention, using the drawing process of CMOS shot records erythrocyte 2.CMOS full name are
Complementary Metal-Oxide-Semicondoctor, Chinese scientific name are complementary metal oxide semiconductors (CMOS), are made with which
Into CMOS camera lenses can quickly process the image that is continually changing, its sensitivity is higher, can realize to nanoparticle image
It is more accurate to determine, it is that measurement result is more accurate reliable.
The embodiment of the present invention also provides a kind of blood oxygen carrying capacity measuring method, and which includes shearing mould using above-mentioned erythrocyte
The measuring method measurement of amount obtains the erythrocyte modulus of shearing of blood to be measured.The erythrocyte modulus of shearing that measurement is obtained is bigger, red
The deformability of cell is less, and the oxygen carrying capacity of blood is poorer.
With reference to embodiments the feature and performance of the present invention are described in further detail.
Embodiment
A kind of measuring method of erythrocyte modulus of shearing that the present embodiment is provided.
S1：The process of sample cell
Coverslip and wave carrier piece are dried using 1% bovine serum albumen solution immersion 20min, using 100 μ m-thick plastic sheets
Pad forms sample room between coverslip and microscope slide.
S2：Sample clean
Experiment silicon dioxide microsphere solution original content is 2.5%w/v, gathers healthy donor's using vacuum test tube
Blood takes 100 μ l silicon dioxide microspheres respectively and 10 μ l blood samples is cleaned as measuring samples.Silicon dioxide microsphere solution and
Blood sample respectively using phosphate buffer (PBS) dilute, silicon dioxide microsphere using be cleaned by ultrasonic 15min, 7000rpm from
Heart 10min is collected, and is repeated 3 times；Erythrocyte is collected using agitation dispersion, 2500rpm centrifugation 10min, is repeated 2 times.
S3：Hatching
The silicon dioxide microsphere for cleaning is dispersed in 0.1ml PBS again, erythrocyte is dispersed in 2.5ml PBS again
In, with mass concentration ratio be 2:1 solution, equal proportion mix homogeneously stand 40min in 4 DEG C of environment and obtain hatching solution.
S4：Addition sample
Hatching solution is diluted into 100 times and is added dropwise to sealed sample pond behind sample room.
S5：Demarcate Optical Trap Stiffness
The silicon dioxide microsphere of erythrocyte is not adhered to using optical tweezer capture, now silicon oxide pellets are only by trapping stiffness
Without any external force.The optical tweezer Optical Trap Stiffness demarcated under different luminous powers using thermal motion analysis method.
It is illustrated in figure 3 the calibration process of Optical Trap Stiffness.Adopt laser power P suspend two are captured for the optical tweezer of 200mW
Silicon oxide microsphere, shown in movement locus of the silicon dioxide microsphere for being captured in ligh trap such as Fig. 3 (a), silicon dioxide microsphere
Shown in position distribution probability such as Fig. 3 (b), fitting obtains ligh trap potential energy, shown in such as Fig. 3 (c), you can obtain Optical Trap Stiffness.Heat fortune
The optical tweezer Optical Trap Stiffness that dynamic analytic process is demarcated under different luminous powers, as a result as shown in Fig. 3 (d), as a result shows, optical tweezer Optical Trap Stiffness
The into good linear relationship with laser power.The result is consistent with the calculated results, shows that above-mentioned scaling method accurately may be used
OK.The optical tweezer Optical Trap Stiffness under different luminous powers can be obtained by linear fit.
S5：Stretching erythrocyte
One of two silicon dioxide microspheres on erythrocyte are adhered to using optical tweezer capture, as shown in Fig. 2 with larger power
Silicon dioxide microsphere is pulled away from sample cell bottom, along the continuously elongated erythrocyte in line direction of two silicon dioxide microspheres, and
With CMOS shot record drawing process；Using image analysis software analysis experiment video.Obtain the relative of trapping stiffness and erythrocyte
Relation curve between elongation；Slope of curve k can be obtained by the linear fit to relation curve, brought into using slope k
Computing formula (4) can draw the shearing modulus of shearing of erythrocyte membrane.
As shown in figure 4, choosing a diameter of 7.70 μm, 6.87 μm, 7.07 μm, 7.25 μm and 7.18 μm of erythrocyte as weight
Multiple measurement object, is carried out continuously elongated respectively with optical tweezer to above-mentioned erythrocyte, obtains five erythrocyte relative elongations and ligh trap
Power relation curve.Above-mentioned relation curve is fitted, obtain relation curve slope k in Fig. 4 be respectively 0.0448,
0.0433rd, 0.03,0.036 and 0.031.
By slope k substitute into formula (4) be calculated erythrocyte modulus of shearing be respectively 7.61 μ N/m, 8.48 μ N/m, 14.48
μ N/m, 10.86 μ N/m and 13.43 μ N/m.Therefore, by the multiple erythrocyte of repeated measure, obtain their average shear modulus
For H=10.95 ± 2.67 μ N/m.The measurement of the same cells that the erythrocyte modulus of shearing that the method is measured is announced with other documents
As a result it is basically identical.
In sum, the measuring method of erythrocyte modulus of shearing provided in an embodiment of the present invention, it is possible to achieve erythrocyte is received
Power area is uniformed, and reduces modification impact of the reagent to measurement result.Erythrocyte is continuously drawn using scanning optical tweezer technology
Stretch, it is not necessary to frequent mobile example platform.Analyze Optical Trap Stiffness and count out using thermal motion analysis method and apply in erythrocyte drawing process
The pulling force being added on cell, operating procedure are few, and operational approach is simple, workable, can greatly shorten erythrocyte shearing
The time of measuring of modulus.Meanwhile, the expense of the measuring method is low, efficiency high, the high precision of measurement result, with good application
Prospect.
Embodiments described above is a part of embodiment of the invention, rather than the embodiment of whole.The reality of the present invention
The detailed description for applying example is not intended to limit the scope of claimed invention, but is merely representative of the selected enforcement of the present invention
Example.Based on the embodiment in the present invention, what those of ordinary skill in the art were obtained under the premise of creative work is not made
Every other embodiment, belongs to the scope of protection of the invention.
Claims (10)
1. a kind of measuring method of erythrocyte modulus of shearing, it is characterised in which includes：
Addition step：Hatching solution is added into sample cell, in the hatching solution, contains erythrocyte and electrolyte microsphere, the electricity
Medium microsphere include the first electrolyte microsphere for adhering to the erythrocyte opposite sides and not with the red blood cell adhesion
Second electrolyte microsphere；
Optical Trap Stiffness demarcating steps：The second electrolyte microsphere is caught using scanning optical tweezer, the ligh trap to the scanning optical tweezer
Rigidity is demarcated；
Stretching step：The first electrolyte microsphere of any side of the erythrocyte is captured using the scanning optical tweezer, continuously
The erythrocyte is stretched, fitting obtains the relative elongation of the erythrocyte and the relation curve of trapping stiffness, the relation curve
Slope be k, wherein, the relative elongation is the diameter of the extended length after the erythrocyte is stretched and the erythrocyte
Ratio, the slope k of the relation curve is the ratio of the relative elongation and the trapping stiffness；
Calculation procedure：The modulus of shearing of the erythrocyte is calculated, computing formula is as follows：
Wherein, H is the modulus of shearing of the erythrocyte, and k is the slope of the relation curve, and d is the diameter of the erythrocyte, B
To distort modulus of shearing, B=2 × 10-19Nm。
2. measuring method according to claim 1, it is characterised in that the scanning optical tweezer is that acousto-optic deflection device scans light
Tweezer.
3. measuring method according to claim 1, it is characterised in that in the Optical Trap Stiffness demarcating steps, the ligh trap
The scaling method of rigidity is thermal motion analysis method.
4. measuring method according to claim 1, it is characterised in that in the stretching step, the relative elongation are led to
Cross image analytical method to measure, the drawing process of the erythrocyte is recorded using image pick-up device, obtain described by image analysis software
Relation curve.
5. measuring method according to claim 1, it is characterised in that in the stretching step, relative along the erythrocyte
The direction of two the first electrolyte microsphere lines of both sides stretches the erythrocyte.
6. measuring method according to claim 1, it is characterised in that in the stretching step, the stretching of the erythrocyte
Speed is less than or equal to 0.02 μm/s.
7. measuring method according to claim 1, it is characterised in that the electrolyte microsphere is the two of a diameter of 3～5 μm
Silicon oxide microsphere.
8. measuring method according to claim 7, it is characterised in that by the silicon dioxide microsphere for cleaning and described
Erythrocyte is prepared and obtains mass concentration ratio for 2:1 mixed solution, hatches the mixed solution and obtains the hatching solution.
9. measuring method according to claim 1, it is characterised in that the bottom of the sample cell has blood stasis albumin
Layer.
10. a kind of blood oxygen carrying capacity measuring method, it is characterised in which is included using such as claim 1～9 any one institute
The measuring method measurement of the erythrocyte modulus of shearing stated obtains the erythrocyte modulus of shearing of blood to be measured.
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| CN110664369A (en) * | 2019-09-19 | 2020-01-10 | 哈尔滨工业大学 | Self-adaptive confocal line scanning harmonic microscopic imaging method and device |
| CN111617390A (en) * | 2020-06-23 | 2020-09-04 | 暨南大学 | Device for regulating and controlling red blood cells in living animal blood vessel and application thereof |
| CN112620113A (en) * | 2020-12-03 | 2021-04-09 | 暨南大学 | Nanoparticle screening and separating device based on scanning type optical tweezers |
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