CN107202773A - A kind of cell cycle scattered light intensity method for establishing model of utilization polystyrene spheres - Google Patents
A kind of cell cycle scattered light intensity method for establishing model of utilization polystyrene spheres Download PDFInfo
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- CN107202773A CN107202773A CN201710404246.9A CN201710404246A CN107202773A CN 107202773 A CN107202773 A CN 107202773A CN 201710404246 A CN201710404246 A CN 201710404246A CN 107202773 A CN107202773 A CN 107202773A
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/47—Scattering, i.e. diffuse reflection
Abstract
The invention provides a kind of cell cycle scattered light intensity method for establishing model of utilization polystyrene spheres, this method is set up by the use of the suspension of different-grain diameter polystyrene spheres and obtained to be used as the cell cycle scattered light intensity model that light-intensity test reference standard is scattered to the living body biological cell cycle, and the property of polystyrene spheres is stable, also will not produce causes the cell cycle to be continually changing because of cell division growth, therefore, it is possible to be able to persistence, the problem of cell cycle scattered light intensity detection reference standard for solving persistence in the prior art is difficult to make；And, when being diluted to the concentration of different gradients due to different-grain diameter polystyrene microsphere, the light scattering characteristic for showing angle dependence light scattering characteristic phase different from the cell cycle is consistent or met after amendment, therefore the cell cycle scattered light intensity model set up using the inventive method has good universality, and improving more cell cycle light intensity of light scattering models for future provides expansible thinking.
Description
Technical field
The present invention relates to technical field of biological, more particularly to Cell Measurement Technique, and in particular to one kind utilizes polyphenyl
The cell cycle scattered light intensity method for establishing model of ethene ball.
Background technology
Cell cycle refers to that cell terminates undergone overall process since being completed once division to division next time, is divided into
Between phase and two stages of division stage.Wherein, a phase is divided into three phases, i.e. DNA pre-synthesis phases again（The G1 phases）, DNA synthesis the phase（The S phases）With
DNA post-synthesis phases（The G2 phases）.Division stage（The M phases）It is divided into early stage, mid-term, later stage and latter stage.
The theoretical cell cycle is not only that studying for tumour provides important theoretical foundation, and the cell cycle is chemotherapy of tumors
Theoretical foundation.Accurately antineoplastic can be killed directly against the cancer cell of different cycles, so as to reach treatment
Purpose.Therefore, treatment of the analysis measurement to cancer is carried out to the cancer cell of different cycles using dependent scattering light intensity technology to examine
It is disconnected significant.
When being detected using scattered light intensity technology to the living body biological cell cycle, its standard scattered light intensity information is built
It is vertical most important.However, because living body biological cell is difficult to permanent survival in culture vessel, and biological cell is in survival
Period also can constantly merisis and cause the cell cycle to be continually changing, therefore be difficult with biological cell and make and preserve each
The scattered light intensity detection reference standard of fixed cell cycle.Other effective cell cycle scattered light intensity models there is no to set up at present
Method.
The content of the invention
For above shortcomings in the prior art, it is an object of the invention to provide a kind of utilization polystyrene spheres
Cell cycle scattered light intensity method for establishing model, it sets up each different cell week using the suspension of different-grain diameter polystyrene spheres
The cell cycle scattered light intensity model of phase, can be to be scattered light intensity as persistence to the living body biological cell cycle
The reference standard of detection, the cell cycle scattered light intensity detection reference standard for solving persistence in the prior art is difficult to what is made
Problem.
In order to solve the above technical problems, present invention employs following technical scheme：
A kind of cell cycle scattered light intensity method for establishing model of utilization polystyrene spheres, pre-production has prescribed concentration not
With the suspension of particle diameter polystyrene spheres, luminous intensity measurement is scattered respectively using wide-angle light scattering apparatus, obtains variant particle diameter
Each self-corresponding scattered light intensity distribution curve of suspension of polystyrene spheres；Then the cell sample of variant cell cycle is used
The cell suspension of the correspondence variant cell cycle with same prescribed concentration is respectively prepared, using wide under identical measuring condition
Angle light scattering apparatus is scattered luminous intensity measurement respectively, obtains each self-corresponding scattered light intensity of cell suspension of each cell cycle
Spend distribution curve, and the scattered light intensity distribution curve of the suspension of variant particle diameter polystyrene spheres with being previously obtained is carried out pair
Than determining the polystyrene that the scattered light intensity distribution curve of the cell suspension of each different cell cycle is each matched respectively
Ball suspension, so as to regard the suspension for matching obtained each particle diameter polystyrene spheres as cell cycle scattered light intensity model.
In the cell cycle scattered light intensity method for establishing model of above-mentioned utilization polystyrene spheres, specifically, specifically include
Following step：
1）Pre-production has the suspension of the different-grain diameter polystyrene spheres of prescribed concentration and loaded in teat glass, shakes up respectively
Afterwards, the suspensions of every kind of particle diameter polystyrene spheres is measured respectively using wide-angle light scattering apparatus in the range of 0 ~ 180 ° at interval of on 5 °
Scattered light intensity, according to measurement result, obtains suspension each self-corresponding scattered light intensity point of variant particle diameter polystyrene spheres
Cloth curve；
2）Using the cell sample of G1 phases, S phases, G2 phases and M phases, the corresponding G1 phases with same prescribed concentration, S is respectively prepared
Phase, the cell suspension of G2 phases and M phases simultaneously load in teat glass, after shaking up respectively, under identical measuring condition, utilize wide-angle light
Scatterometer measures the cell suspension of each cell cycle in the range of 0 ~ 180 ° at interval of the scattered light intensity on 5 ° respectively, according to
Measurement result, obtains each self-corresponding scattered light intensity distribution curve of cell suspension of G1 phases, S phases, G2 phases and M phases；
3）By the cell suspension of G1 phases, S phases, G2 phases and M phases each self-corresponding scattered light intensity distribution curve and it is previously obtained
The scattered light intensity distribution curve of the suspension of variant particle diameter polystyrene spheres is contrasted, and G1 phases, S phases, G2 phases are determined respectively
The polystyrene spheres suspension each matched with the scattered light intensity distribution curve of the cell suspension of M phases, so that matching be obtained
The suspensions of each particle diameter polystyrene spheres be used as cell cycle scattered light intensity model.
It is preferably, described in the cell cycle scattered light intensity method for establishing model of above-mentioned utilization polystyrene spheres
Step 1）In, when measuring the scattered light intensity distribution curve of suspension of every kind of particle diameter polystyrene spheres, duplicate measurements 5 ~ 10 times
The suspension of corresponding particle diameter polystyrene spheres, at interval of the scattered light intensity on 5 °, rejects wherein deviation from average rate in the range of 0 ~ 180 °
Absolute value be more than 10% scattered light intensity angle value, then the scattered light intensity angle value obtained by other time measurement on the scattering angle is asked
It is average, scattered light intensity average value of the suspension of the particle diameter polystyrene spheres on corresponding firing angle degree is obtained, so as to respectively obtain
The suspension of the particle diameter polystyrene spheres, at interval of the scattered light intensity average value on 5 °, is gone forward side by side in 0 ° ~ 180 ° range of scatter angles
Row curve data fit and the disposal of gentle filter, obtain the scattered light intensity distribution corresponding to the suspension of the particle diameter polystyrene spheres
Curve.
It is preferably, described in the cell cycle scattered light intensity method for establishing model of above-mentioned utilization polystyrene spheres
Step 2）In, when measuring the scattered light intensity distribution curve of cell suspension of each cell cycle, duplicate measurements 5 ~ 10 times should
The cell suspension of cell cycle at interval of the scattered light intensity on 5 °, rejects the absolute of wherein deviation from average rate in the range of 0 ~ 180 °
Scattered light intensity angle value of the value more than 10%, then the scattered light intensity angle value obtained by other measurements on the scattering angle is averaging, obtain
To scattered light intensity average value of the cell suspension on corresponding firing angle degree of the cell cycle, so as to respectively obtain the cell cycle
Cell suspension in 0 ° ~ 180 ° range of scatter angles at interval of the scattered light intensity average value on 5 °, and carry out curve data plan
Close and the disposal of gentle filter, finally obtain the scattered light intensity distribution curve corresponding to the cell suspension of the cell cycle.
It is preferably, made in the cell cycle scattered light intensity method for establishing model of above-mentioned utilization polystyrene spheres
Into polystyrene spheres suspension prescribed concentration be 106Individual polystyrene spheres/mL, 2 × 105Individual polystyrene spheres/mL, 105It is individual
Polystyrene spheres/mL or 5 × 104Individual polystyrene spheres/mL, the prescribed concentration of made cell suspension is corresponding for 106It is individual
Cell/mL, 2 × 105Individual cell/mL, 105Individual cell/mL or 5 × 104Individual cell/mL.
In the cell cycle scattered light intensity method for establishing model of above-mentioned utilization polystyrene spheres, preferably, utilize
When wide-angle light scattering apparatus is scattered luminous intensity measurement, light source is used as using laser.
It is preferably, described in the cell cycle scattered light intensity method for establishing model of above-mentioned utilization polystyrene spheres
The optical maser wavelength of laser is 532nm.
Compared to prior art, the present invention has the advantages that：
1st, the present invention can utilize different-grain diameter polyphenyl using the cell cycle scattered light intensity method for establishing model of polystyrene spheres
The suspension of ethene ball is set up and obtained to as the cell that light-intensity test reference standard is scattered to the living body biological cell cycle
Cycle scattered light intensity model, and the property of polystyrene spheres is stable, will not also produce causes cell week because of cell division growth
Phase is continually changing, therefore, it is possible to be able to persistence, solves the cell cycle scattered light intensity inspection of persistence in the prior art
Survey the problem of reference standard is difficult to make.
When the 2nd, being diluted to the concentration of different gradients due to different-grain diameter polystyrene microsphere, show angle and rely on light scattering
The light scattering characteristic of characteristic phase different from the cell cycle is consistent or met after amendment, therefore uses the inventive method
The cell cycle scattered light intensity model of foundation has good universality, is to improve more cell cycle light scattering light future
Strong model provides expansible thinking, with good technology application prospect.
Brief description of the drawings
Fig. 1 is flow chart of the present invention using the cell cycle scattered light intensity method for establishing model of polystyrene spheres.
Fig. 2 is G1 phases, S phases, the cell suspension scattered light intensity point of four different cell cycles of G2 phases and M phases in embodiment
Cloth curve map.
Fig. 3 be embodiment in the G1 phases HeLa cell suspensions and 12 μm of particle diameter polystyrene spheres suspension scattered light intensity
Distribution curve comparison diagram.
Fig. 4 be embodiment in the S phases HeLa cell suspensions and 14 μm of particle diameter polystyrene spheres suspension scattered light intensity
Distribution curve comparison diagram.
Fig. 5 be embodiment in the G2 phases HeLa cell suspensions and 11 μm of particle diameter polystyrene spheres suspension scattered light intensity
Distribution curve comparison diagram.
Fig. 6 be embodiment in the M phases HeLa cell suspensions and 22 μm of particle diameter polystyrene spheres suspension scattered light intensity
Distribution curve comparison diagram.
Embodiment
The problem of for how to set up cell cycle scattered light intensity model, find that polystyrene microsphere is good by research
Can be to set up the material of cell cycle scattered light intensity model.Polystyrene microsphere is to carry out addition by styrene monomer to gather
Formed after conjunction, feature is colourless, transparent, and optical property is fabulous, is verified by repetitive measuring experiment, polystyrene spheres
Scattered light intensity distribution curve it is stable, repetitive rate is high；Meanwhile, polystyrene microsphere has good heat resistance, and it is entered
Row sterilization treatment does not interfere with its performance yet；In addition, the dispersive property of polystyrene microsphere in aqueous is preferable, it is not easy to
Reunite.These characteristics that polystyrene spheres possess, the important spy of analog cell scattered light intensity can be used to by embodying it
Property, it is the preferable selection as cell scattered light intensity model.
Based on this, the invention provides a kind of cell cycle scattered light intensity method for establishing model of utilization polystyrene spheres,
Embodiments thereof is that pre-production has the suspension of the different-grain diameter polystyrene spheres of prescribed concentration, utilizes wide-angle light scattering apparatus
Luminous intensity measurement is scattered respectively, obtains each self-corresponding scattered light intensity distribution of suspension of variant particle diameter polystyrene spheres
Curve；Then the variant cell of correspondence with same prescribed concentration is respectively prepared using the cell sample of variant cell cycle
The cell suspension in cycle, luminous intensity measurement is scattered under identical measuring condition respectively using wide-angle light scattering apparatus, obtains each
Each self-corresponding scattered light intensity distribution curve of cell suspension of individual cell cycle, and with the variant particle diameter polyphenyl that is previously obtained
The scattered light intensity distribution curve of the suspension of ethene ball is contrasted, and the cell suspension of each different cell cycle is determined respectively
The polystyrene spheres suspension that scattered light intensity distribution curve is each matched, so that obtained each particle diameter polystyrene spheres will be matched
Suspension be used as cell cycle scattered light intensity model.
Specifically, the present invention utilizes cell cycle scattered light intensity method for establishing model flow such as Fig. 1 of polystyrene spheres
It is shown, comprise the steps：
1）Pre-production has the suspension of the different-grain diameter polystyrene spheres of prescribed concentration and loaded in teat glass, shakes up respectively
Afterwards, the suspensions of every kind of particle diameter polystyrene spheres is measured respectively using wide-angle light scattering apparatus in the range of 0 ~ 180 ° at interval of on 5 °
Scattered light intensity, according to measurement result, obtains suspension each self-corresponding scattered light intensity point of variant particle diameter polystyrene spheres
Cloth curve.
Measurement obtains each self-corresponding scattered light intensity distribution song of suspension of variant particle diameter polystyrene spheres respectively in advance
Line, can be used for matching and makes the cell cycle scattered light intensity model for being directed to variety classes cell.Specifically measuring every kind of particle diameter
, can be with 5 ~ 10 corresponding particle diameter polystyrene spheres of duplicate measurements during the scattered light intensity distribution curve of the suspension of polystyrene spheres
Suspension, at interval of the scattered light intensity on 5 °, rejects the absolute value of wherein deviation from average rate dissipating more than 10% in the range of 0 ~ 180 °
Light intensity value is penetrated, then the scattered light intensity angle value obtained by other measurements on the scattering angle is averaging, the particle diameter polyphenyl is obtained
Scattered light intensity average value of the suspension of ethene ball on corresponding firing angle degree, so as to respectively obtain the outstanding of the particle diameter polystyrene spheres
Liquid is interior at interval of the scattered light intensity average value on 5 ° in 0 ° ~ 180 ° range of scatter angles, and carries out curve data fit and smooth
Filtering process, finally obtains the scattered light intensity distribution curve corresponding to the suspension of the particle diameter polystyrene spheres.It can so disappear
Except scattered light intensity distortion error caused by a variety of causes in measurement, the scattered light of the polystyrene spheres suspension of gained is preferably ensured
The measurement accuracy of strength distribution curve.
2）Using the cell sample of G1 phases, S phases, G2 phases and M phases, the corresponding G1 with same prescribed concentration is respectively prepared
Phase, S phases, the cell suspension of G2 phases and M phases simultaneously load in teat glass, after shaking up respectively, under identical measuring condition, using wide
Angle light scattering apparatus measures the cell suspension of each cell cycle in the range of 0 ~ 180 ° at interval of the scattered light intensity on 5 ° respectively,
According to measurement result, each self-corresponding scattered light intensity distribution curve of cell suspension of G1 phases, S phases, G2 phases and M phases are obtained.
When specifically measuring the scattered light intensity distribution curve of the cell suspension of each cell cycle, can with duplicate measurements 5 ~
The cell suspension of 10 cell cycles, at interval of the scattered light intensity on 5 °, rejects wherein deviation from average rate in the range of 0 ~ 180 °
Absolute value be more than 10% scattered light intensity angle value, then the scattered light intensity angle value obtained by other time measurement on the scattering angle is asked
It is average, scattered light intensity average value of the cell suspension of the cell cycle on corresponding firing angle degree is obtained, so as to respectively obtain this
The cell suspension of cell cycle in 0 ° ~ 180 ° range of scatter angles at interval of the scattered light intensity average value on 5 °, and march
Line number is according to fitting and the disposal of gentle filter, and the scattered light intensity distribution finally obtained corresponding to the cell suspension of the cell cycle is bent
Line.Scattered light intensity distortion error caused by a variety of causes in measurement can be so eliminated, the cell suspension of gained is preferably ensured
Scattered light intensity distribution curve measurement accuracy.
3）Each self-corresponding scattered light intensity distribution curve of the cell suspension of G1 phases, S phases, G2 phases and M phases is obtained with advance
To the scattered light intensity distribution curves of suspension of variant particle diameter polystyrene spheres contrasted, determine respectively the G1 phases, the S phases,
The polystyrene spheres suspension that the scattered light intensity distribution curve of the cell suspension of G2 phases and M phases is each matched, so that will matching
The suspension of obtained each particle diameter polystyrene spheres is as cell cycle scattered light intensity model.
It can see by above-mentioned flow, the present invention utilizes the cell cycle scattered light intensity model foundation side of polystyrene spheres
Method, can be set up by the use of the suspension of different-grain diameter polystyrene spheres and obtained to be scattered as to the living body biological cell cycle
The cell cycle scattered light intensity model of light-intensity test reference standard, and the property of polystyrene spheres is stable, will not also produce because thin
Born of the same parents' merisis and cause the cell cycle to be continually changing, therefore, it is possible to be able to persistence, solve permanent guarantor in the prior art
The problem of cell cycle scattered light intensity detection reference standard deposited is difficult to make.
In the specific implementation, the prescribed concentration of the suspension of made polystyrene spheres can be 106Individual polystyrene spheres/
mL、2×105Individual polystyrene spheres/mL, 105Individual polystyrene spheres/mL or 5 × 104Individual these four concentration of polystyrene spheres/mL,
The prescribed concentration of made cell suspension can also be correspondingly 106Individual cell/mL, 2 × 105Individual cell/mL, 105It is individual thin
Born of the same parents/mL or 5 × 104Individual these four concentration of cell/mL, because the polystyrene spheres suspension and cell of this four concentration ranges hang
Liquid, multiple scattering will not be impacted to scattered light intensity, and scattered light intensity can be caused very weak if concentration is too low, be unfavorable for surveying
Amount, therefore these four concentration are selected as prescribed concentration, it is advantageously ensured that determining the repetition of cell cycle based on light scattering measurement
Property and stability.Wherein, as most preferably scheme, the setting of prescribed concentration is preferably 105Individual polystyrene spheres/mL, 105It is individual thin
Born of the same parents/mL, its benefit is that the scattered light intensity under the concentration is moderate, can conveniently observe scattered light between different samples
Strong difference., can be poly- as reagent dilutions using deionized water or trypsin solution when making the suspension of polystyrene spheres
Styrene ball is to prescribed concentration；And when making cell supernatant liquid, reagent dilutions cell sample can be used as using trypsin solution
To prescribed concentration.And when being scattered luminous intensity measurement using wide-angle light scattering apparatus, preferably with laser as light source, its
Advantage is good beam quality, with extraordinary monochromaticjty, directionality and stability.In addition, the optical maser wavelength of laser is best
It is set as 532nm, because cell scattered light intensity difference at that wavelength is optimal, and the performance of dynamic light scattering and measurement are tied
Fruit is more preferable at that wavelength.
Illustrate the inventive method with embodiment below.It should be understood that these embodiments are only used for into one
Step explanation embodiment of the present invention, rather than for limiting the present invention.
Embodiment：
Using the method for the present invention, the cell cycle scattered light intensity model of HeLa cells is set up, following step is specifically included：
1）Pre-production prescribed concentration is 105The suspension of individual polystyrene spheres/mL different-grain diameter polystyrene spheres simultaneously loads glass
In glass test tube, after shaking up respectively, the suspension of every kind of particle diameter polystyrene spheres is measured respectively at 0 ~ 180 ° using wide-angle light scattering apparatus
In the range of at interval of the scattered light intensity on 5 °, using laser as light source, and the optical maser wavelength of laser is 532nm, root
According to measurement result, each self-corresponding scattered light intensity distribution curve of suspension of variant particle diameter polystyrene spheres is obtained.
2）Using the HeLa cell samples of G1 phases, S phases, G2 phases and M phases, respectively by 105Individual cell/mL prescribed concentration system
Into correspondence G1 phases, S phases, the HeLa cell suspensions of G2 phases and M phases and load in teat glass, after shaking up respectively, in identical measurement
Under the conditions of, using wide-angle light scattering apparatus measure respectively the HeLa cell suspensions of each cell cycle in the range of 0 ~ 180 ° at interval of
Scattered light intensity on 5 °, using laser as light source, and the optical maser wavelength of laser is 532nm, according to measurement result, is obtained
To each self-corresponding scattered light intensity distribution curve of HeLa cell suspensions of G1 phases, S phases, G2 phases and M phases.
In the present embodiment, resulting G1 phases, S phases, each self-corresponding scattered light intensity of the HeLa cell suspensions of G2 phases and M phases
Spend distribution curve as shown in Figure 2.
3）By each self-corresponding scattered light intensity distribution curve of the HeLa cell suspensions of G1 phases, S phases, G2 phases and M phases and in advance
The scattered light intensity distribution curve of the suspension of the variant particle diameter polystyrene spheres first obtained is contrasted, and G1 phases, S are determined respectively
The polystyrene spheres suspension that the scattered light intensity distribution curve of the HeLa cell suspensions of phase, G2 phases and M phases is each matched, so that
The suspension of obtained each particle diameter polystyrene spheres will be matched as the cell cycle scattered light intensity model of HeLa cells.
In the present embodiment, learnt by contrast, the HeLa cell suspensions of G1 phases（G1 phase cell）It is poly- with 12 μm of particle diameters
The suspension of styrene ball（12μm PS）Scattered light intensity distribution curve match, as shown in Figure 3；The HeLa cell suspensions of S phases
（S phase cell）With the suspension of 14 μm of particle diameter polystyrene spheres（14μm PS）Scattered light intensity distribution curve match,
As shown in Figure 4；The HeLa cell suspensions of G2 phases（G2 phase cell）With the suspension of 11 μm of particle diameter polystyrene spheres（11μm
PS）Scattered light intensity distribution curve match, as shown in Figure 5；And the HeLa cell suspensions of M phases（M phase cell）With 22
The suspension of μm particle diameter polystyrene spheres（22μm PS）Scattered light intensity distribution curve match, as shown in Figure 6.Thus, just divide
Not using the suspension of 12 μm, 14 μm, 11 μm and 22 μm particle diameter polystyrene spheres as HeLa cells in G1 phases, S phases, G2 phases and M phases
Cell cycle scattered light intensity model.
In above-mentioned Fig. 2 ~ Fig. 6, abscissa represents scatter angle values（angle）, ordinate represents scattered light intensity
Angle value（light scattering）.
In summary, the present invention can be utilized using the cell cycle scattered light intensity method for establishing model of polystyrene spheres
The suspensions of different-grain diameter polystyrene spheres, which is set up, to be obtained to as being scattered light-intensity test ginseng to the living body biological cell cycle
The accurate cell cycle scattered light intensity model of sighting target, and the property of polystyrene spheres is stable, will not also produce because of cell division growth
And cause the cell cycle to be continually changing, therefore, it is possible to be able to persistence, solve the cell week of persistence in the prior art
The problem of phase scattered light intensity detection reference standard is difficult to make；Also, because different-grain diameter polystyrene microsphere is diluted to difference
During the concentration of gradient, show angle rely on different from the cell cycle phase of light scattering characteristic light scattering characteristic be consistent or
Meet after amendment, therefore the cell cycle scattered light intensity model set up using the inventive method has good universality,
More cell cycle light intensity of light scattering models will be improved for future and provide expansible thinking, with good technology application
Prospect.
Finally illustrate, the above embodiments are merely illustrative of the technical solutions of the present invention and it is unrestricted, although with reference to compared with
The present invention is described in detail good embodiment, it will be understood by those within the art that, can be to skill of the invention
Art scheme is modified or equivalent substitution, and without departing from the objective and scope of technical solution of the present invention, it all should cover at this
Among the right of invention.
Claims (7)
1. the cell cycle scattered light intensity method for establishing model of a kind of utilization polystyrene spheres, it is characterised in that pre-production has
There is the suspension of the different-grain diameter polystyrene spheres of prescribed concentration, luminous intensity measurement be scattered respectively using wide-angle light scattering apparatus,
Obtain each self-corresponding scattered light intensity distribution curve of suspension of variant particle diameter polystyrene spheres；Then variant cell is used
The cell suspension of the correspondence variant cell cycle with same prescribed concentration is respectively prepared in the cell sample in cycle, is surveyed identical
Luminous intensity measurement is scattered respectively using wide-angle light scattering apparatus under the conditions of amount, obtains the cell suspension of each cell cycle each
Corresponding scattered light intensity distribution curve, and with the scattered light intensity for the suspension of variant particle diameter polystyrene spheres being previously obtained
Distribution curve is contrasted, and the respective institute of scattered light intensity distribution curve of the cell suspension of each different cell cycle is determined respectively
The polystyrene spheres suspension of matching, so as to regard the suspension for matching obtained each particle diameter polystyrene spheres as cell cycle scattered light
Strong model.
2. utilizing the cell cycle scattered light intensity method for establishing model of polystyrene spheres according to claim 1, its feature exists
In specifically including following step：
1）Pre-production has the suspension of the different-grain diameter polystyrene spheres of prescribed concentration and loaded in teat glass, shakes up respectively
Afterwards, the suspensions of every kind of particle diameter polystyrene spheres is measured respectively using wide-angle light scattering apparatus in the range of 0 ~ 180 ° at interval of on 5 °
Scattered light intensity, according to measurement result, obtains suspension each self-corresponding scattered light intensity point of variant particle diameter polystyrene spheres
Cloth curve；
2）Using the cell sample of G1 phases, S phases, G2 phases and M phases, the corresponding G1 phases with same prescribed concentration, S is respectively prepared
Phase, the cell suspension of G2 phases and M phases simultaneously load in teat glass, after shaking up respectively, under identical measuring condition, utilize wide-angle light
Scatterometer measures the cell suspension of each cell cycle in the range of 0 ~ 180 ° at interval of the scattered light intensity on 5 ° respectively, according to
Measurement result, obtains each self-corresponding scattered light intensity distribution curve of cell suspension of G1 phases, S phases, G2 phases and M phases；
3）By the cell suspension of G1 phases, S phases, G2 phases and M phases each self-corresponding scattered light intensity distribution curve and it is previously obtained
The scattered light intensity distribution curve of the suspension of variant particle diameter polystyrene spheres is contrasted, and G1 phases, S phases, G2 phases are determined respectively
The polystyrene spheres suspension each matched with the scattered light intensity distribution curve of the cell suspension of M phases, so that matching be obtained
The suspensions of each particle diameter polystyrene spheres be used as cell cycle scattered light intensity model.
3. utilizing the cell cycle scattered light intensity method for establishing model of polystyrene spheres according to claim 2, its feature exists
In the step 1）In, when measuring the scattered light intensity distribution curve of suspension of every kind of particle diameter polystyrene spheres, duplicate measurements
The suspension of 5 ~ 10 corresponding particle diameter polystyrene spheres in the range of 0 ~ 180 ° at interval of the scattered light intensity on 5 °, reject wherein from
The absolute value of equal rate is more than 10% scattered light intensity angle value, then by the scattered light intensity obtained by other measurements on the scattering angle
Angle value is averaging, and obtains scattered light intensity average value of the suspension of the particle diameter polystyrene spheres on corresponding firing angle degree, so that point
The suspension for not obtaining the particle diameter polystyrene spheres is averaged in 0 ° ~ 180 ° range of scatter angles at interval of the scattered light intensity on 5 °
Value, and curve data fit and the disposal of gentle filter are carried out, obtain the scattered light corresponding to the suspension of the particle diameter polystyrene spheres
Strength distribution curve.
4. utilizing the cell cycle scattered light intensity method for establishing model of polystyrene spheres according to claim 2, its feature exists
In the step 2）In, when measuring the scattered light intensity distribution curve of cell suspension of each cell cycle, duplicate measurements 5 ~
The cell suspension of 10 cell cycles, at interval of the scattered light intensity on 5 °, rejects wherein deviation from average rate in the range of 0 ~ 180 °
Absolute value be more than 10% scattered light intensity angle value, then the scattered light intensity angle value obtained by other time measurement on the scattering angle is asked
It is average, scattered light intensity average value of the cell suspension of the cell cycle on corresponding firing angle degree is obtained, so as to respectively obtain this
The cell suspension of cell cycle in 0 ° ~ 180 ° range of scatter angles at interval of the scattered light intensity average value on 5 °, and march
Line number is according to fitting and the disposal of gentle filter, and the scattered light intensity distribution finally obtained corresponding to the cell suspension of the cell cycle is bent
Line.
5. utilizing the cell cycle scattered light intensity method for establishing model of polystyrene spheres according to claim 1, its feature exists
In the prescribed concentration of the suspension of made polystyrene spheres is 106Individual polystyrene spheres/mL, 2 × 105Individual polystyrene spheres/
mL、105Individual polystyrene spheres/mL or 5 × 104Individual polystyrene spheres/mL, the prescribed concentration of made cell suspension is corresponding
For 106Individual cell/mL, 2 × 105Individual cell/mL, 105Individual cell/mL or 5 × 104Individual cell/mL.
6. utilizing the cell cycle scattered light intensity method for establishing model of polystyrene spheres according to claim 1, its feature exists
In when being scattered luminous intensity measurement using wide-angle light scattering apparatus, light source is used as using laser.
7. utilizing the cell cycle scattered light intensity method for establishing model of polystyrene spheres according to claim 4, its feature exists
In the optical maser wavelength of the laser is 532nm.
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