CN107202773B - A kind of cell cycle scattered light intensity method for establishing model using polystyrene spheres - Google Patents
A kind of cell cycle scattered light intensity method for establishing model using polystyrene spheres Download PDFInfo
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- CN107202773B CN107202773B CN201710404246.9A CN201710404246A CN107202773B CN 107202773 B CN107202773 B CN 107202773B CN 201710404246 A CN201710404246 A CN 201710404246A CN 107202773 B CN107202773 B CN 107202773B
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- 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 present invention provides a kind of cell cycle scattered light intensity method for establishing model using polystyrene spheres, this method is established to obtain to as the cell cycle scattered light intensity model for being scattered the living body biological cell cycle light-intensity test reference standard using the suspension of different-grain diameter polystyrene spheres, and the property of polystyrene spheres is stablized, will not generate causes the cell cycle constantly to change because of cell division growth, therefore it can be able to persistence, solves the problems, such as that the cell cycle scattered light intensity detection reference standard of 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 meets after amendment, therefore there is good universality using the cell cycle scattered light intensity model that the method for the present invention is established, will improve more cell cycle light intensity of light scattering models for future and provides expansible thinking.
Description
Technical field
The present invention relates to technical field of biological more particularly to Cell Measurement Techniques, and in particular to a kind of to utilize polyphenyl
The cell cycle scattered light intensity method for establishing model of ethylene ball.
Background technique
Cell cycle refers to that cell terminates overall process experienced to division next time since being completed primary division, is divided into
Interphase and two stages of division stage.Wherein, interphase be divided into again three phases, i.e. DNA pre-synthesis phase (G1 phase), DNA synthesis phase (S phase) with
DNA post-synthesis phase (G2 phase).Division stage (M phase), is divided into early period, mid-term, later period and latter stage.
Cell cycle theory is not only that the research of tumour provides important theoretical foundation, and the cell cycle is chemotherapy of tumors
Theoretical basis.Accurately anti-tumor drug can be killed directly against the cancer cell of different cycles, to reach treatment
Purpose.Therefore, analysis measurement is carried out using cancer cell of the dependent scattering light intensity technology to different cycles to examine the treatment of cancer
It is disconnected to be of great significance.
When being detected using scattered light intensity technology to the living body biological cell cycle, standard scattered light intensity information is built
It stands most important.However, since living body biological cell is difficult to permanent survival in culture vessel, and biological cell is being survived
Period also can constantly merisis and cause the cell cycle constantly to change, therefore be difficult with biological cell production and save each
The scattered light intensity of fixed cell cycle detects reference standard.It there is no other effective cell cycle scattered light intensity model foundations at present
Method.
Summary of the invention
Aiming at the above shortcomings existing in the prior art, the purpose of the present invention is to provide a kind of using polystyrene spheres
Cell cycle scattered light intensity method for establishing model establishes each different cell weeks 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 to the living body biological cell cycle as persistence
The reference standard of detection solves what the cell cycle scattered light intensity detection reference standard of persistence in the prior art was difficult to make
Problem.
In order to solve the above technical problems, present invention employs the following technical solutions:
A kind of cell cycle scattered light intensity method for establishing model using polystyrene spheres, pre-production have prescribed concentration
Different-grain diameter polystyrene spheres suspension, be scattered luminous intensity measurement respectively using wide-angle light scattering apparatus, obtain variant
The corresponding scattered light intensity distribution curve of the suspension of partial size polystyrene spheres；Then the cell of variant cell cycle is used
The cell suspension of the correspondence variant cell cycle with same prescribed concentration is respectively prepared in sample, sharp under identical measuring condition
It is scattered luminous intensity measurement respectively with wide-angle light scattering apparatus, obtains the corresponding scattering of cell suspension of each cell cycle
Light intensity distributions curve, and with the scattered light intensity distribution curve of the suspension for the variant partial size polystyrene spheres being previously obtained into
Row comparison, determine respectively the scattered light intensity distribution curve of the cell suspension of each different cell cycle respectively matched polyphenyl
Ethylene ball suspension, so that the suspension for each partial size polystyrene spheres that matching is obtained is as cell cycle scattered light intensity model.
In the above-mentioned cell cycle scattered light intensity method for establishing model using polystyrene spheres, specifically, specifically including
Following step:
1) pre-production has the suspension of the different-grain diameter polystyrene spheres of prescribed concentration and is packed into teat glass, respectively
After shaking up, the suspensions of every kind of partial size polystyrene spheres is measured respectively within the scope of 0 ~ 180 ° at interval of 5 ° using wide-angle light scattering apparatus
On scattered light intensity the corresponding scattered light intensity of suspension of variant partial size polystyrene spheres is obtained according to measurement result
Spend distribution curve；
2) using the cell sample of G1 phase, S phase, G2 phase and M phase, the correspondence G1 with same prescribed concentration is respectively prepared
Phase, the S phase, G2 phase and M phase cell suspension and be packed into teat glass, after shaking up respectively, under identical measuring condition, using wide
Angle light scattering apparatus measures the cell suspension of each cell cycle within the scope of 0 ~ 180 ° at interval of the scattered light intensity on 5 ° respectively,
According to measurement result, the corresponding scattered light intensity distribution curve of cell suspension of G1 phase, S phase, G2 phase and M phase are obtained；
3) the corresponding scattered light intensity distribution curve of the cell suspension of G1 phase, S phase, G2 phase and M phase is obtained with preparatory
To the scattered light intensity distribution curves of suspension of variant partial size polystyrene spheres compare, determine respectively the G1 phase, the S phase,
The respective matched polystyrene spheres suspension of institute of the scattered light intensity distribution curve of the cell suspension of G2 phase and M phase, thus will matching
The suspension of obtained each partial size polystyrene spheres is as cell cycle scattered light intensity model.
It is preferably, described in the above-mentioned cell cycle scattered light intensity method for establishing model using polystyrene spheres
In step 1), when measuring the scattered light intensity distribution curve of suspension of every kind of partial size polystyrene spheres, duplicate measurements 5 ~ 10 times
The suspension of corresponding partial size polystyrene spheres, at interval of the scattered light intensity on 5 °, rejects wherein deviation from average rate within the scope of 0 ~ 180 °
Absolute value be greater than 10% scattered light intensity angle value, then the resulting scattered light intensity angle value of measurements other time on the scattering angle are asked
It is average, scattered light intensity average value of the suspension of the partial size polystyrene spheres on corresponding firing angle degree is obtained, to respectively obtain
The suspension of the partial size 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 distribution of scattered light intensity corresponding to the suspension of the partial size polystyrene spheres
Curve.
It is preferably, described in the above-mentioned cell cycle scattered light intensity method for establishing model using polystyrene spheres
In step 2, 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 within the scope of 0 ~ 180 °
Value is greater than 10% scattered light intensity angle value, then the other resulting scattered light intensity angle value of measurement on the scattering angle are averaging, and obtains
To scattered light intensity average value of the cell suspension on corresponding firing angle degree of the cell cycle, 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 it is quasi- to carry out curve data
Conjunction and the disposal of gentle filter, finally obtain scattered light intensity distribution curve corresponding to the cell suspension of the cell cycle.
It is preferably, made in the above-mentioned cell cycle scattered light intensity method for establishing model using polystyrene spheres
At polystyrene spheres suspension prescribed concentration be 106A polystyrene spheres/mL, 2 × 105A polystyrene spheres/mL, 105It is a
Polystyrene spheres/mL or 5 × 104A polystyrene spheres/mL, the corresponding prescribed concentration of made cell suspension is 106It is a
Cell/mL, 2 × 105A cell/mL, 105A cell/mL or 5 × 104A cell/mL.
In the above-mentioned cell cycle scattered light intensity method for establishing model using polystyrene spheres, preferably, utilize
When wide-angle light scattering apparatus is scattered luminous intensity measurement, using laser as light source.
It is preferably, described in the above-mentioned cell cycle scattered light intensity method for establishing model using polystyrene spheres
The optical maser wavelength of laser is 532nm.
Compared with the prior art, the invention has the following beneficial effects:
1, the present invention utilizes the cell cycle scattered light intensity method for establishing model of polystyrene spheres, can utilize different-grain diameter
The suspension of polystyrene spheres is established to obtain to be scattered light-intensity test reference standard as to the living body biological cell cycle
Cell cycle scattered light intensity model, and the property of polystyrene spheres is stablized, will not generate causes carefully because of cell division growth
Born of the same parents constantly change in the period, therefore can be able to persistence, solve the cell cycle scattering light of persistence in the prior art
Strong detection reference standard is difficult to the problem of making.
2, it when being diluted to the concentration of different gradients due to different-grain diameter polystyrene microsphere, shows angle and relies on light scattering
The light scattering characteristic of characteristic phase different from the cell cycle is consistent or meets after amendment, therefore uses the method for the present invention
The cell cycle scattered light intensity model of foundation has good universality, will improve more cell cycle light for future and scatters light
Strong model provides expansible thinking, has good technical application prospect.
Detailed description of the invention
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 the cell suspension scattered light intensity point of G1 phase, S phase, G2 phase and the M phase four different cell cycles in embodiment
Cloth curve graph.
Fig. 3 is the scattered light intensity of the suspension of the HeLa cell suspension and 12 μm of partial size polystyrene spheres of G1 phase in embodiment
Distribution curve comparison diagram.
Fig. 4 is the scattered light intensity of the suspension of the HeLa cell suspension and 14 μm of partial size polystyrene spheres of S phase in embodiment
Distribution curve comparison diagram.
Fig. 5 is the scattered light intensity of the suspension of the HeLa cell suspension and 11 μm of partial size polystyrene spheres of G2 phase in embodiment
Distribution curve comparison diagram.
Fig. 6 is the scattered light intensity of the suspension of the HeLa cell suspension and 22 μm of partial size polystyrene spheres of M phase in embodiment
Distribution curve comparison diagram.
Specific embodiment
It aiming at the problem that how to establish cell cycle scattered light intensity model, finds after study, polystyrene microsphere is very
It is good can be to establish the material of cell cycle scattered light intensity model.Polystyrene microsphere is added by styrene monomer
At what is formed after polymerization, feature is colourless, transparent, and optical property is fabulous, verifies by repetitive measuring experiment, polyphenyl second
The scattered light intensity distribution curve of alkene ball is stablized, and repetitive rate is high；Meanwhile polystyrene microsphere has good heat resistance, it is right
It, which carries out sterilization treatment, will not influence its performance；In addition, the dispersion performance of polystyrene microsphere in aqueous solution is preferable, no
It is easy to happen reunion.These characteristics that polystyrene spheres have, embodying it can be to simulate the weight of cell scattered light intensity
Characteristic is wanted, is the preferable selection as cell scattered light intensity model.
Based on this, the present invention provides a kind of cell cycle scattered light intensity method for establishing model using polystyrene spheres,
Embodiments thereof are that pre-production has the suspension of the different-grain diameter polystyrene spheres of prescribed concentration, utilize wide-angle light scattering apparatus
It is scattered luminous intensity measurement respectively, obtains the corresponding scattered light intensity distribution of suspension of variant partial size 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 period is scattered luminous intensity measurement using wide-angle light scattering apparatus under identical measuring condition respectively, obtains each
The corresponding scattered light intensity distribution curve of the cell suspension of a cell cycle, and with the variant partial size polyphenyl that is previously obtained
The scattered light intensity distribution curve of the suspension of ethylene ball compares, and determines the cell suspension of each different cell cycles respectively
The respective matched polystyrene spheres suspension of institute of scattered light intensity distribution curve, thus each partial size polystyrene spheres that matching is obtained
Suspension as cell cycle scattered light intensity model.
Specifically, the present invention utilizes cell cycle scattered light intensity method for establishing model process such as Fig. 1 of polystyrene spheres
It is shown, include the following steps:
1) pre-production has the suspension of the different-grain diameter polystyrene spheres of prescribed concentration and is packed into teat glass, respectively
After shaking up, the suspensions of every kind of partial size polystyrene spheres is measured respectively within the scope of 0 ~ 180 ° at interval of 5 ° using wide-angle light scattering apparatus
On scattered light intensity the corresponding scattered light intensity of suspension of variant partial size polystyrene spheres is obtained according to measurement result
Spend distribution curve.
Measurement obtains the corresponding scattered light intensity distribution song of suspension of variant partial size polystyrene spheres respectively in advance
Line can be used for matching the cell cycle scattered light intensity model that production is directed to variety classes cell.Specifically measuring every kind of partial size
It, can be with duplicate measurements 5 ~ 10 times corresponding partial size polystyrene spheres when 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 greater than 10% within the scope of 0 ~ 180 °
Light intensity value is penetrated, then the other resulting scattered light intensity angle value of measurement on the scattering angle are averaging, obtains the partial size polyphenyl
Scattered light intensity average value of the suspension of ethylene ball on corresponding firing angle degree, to respectively obtain the outstanding of the partial size polystyrene spheres
Liquid at interval of the scattered light intensity average value on 5 °, and carries out curve data fit and smooth in 0 ° ~ 180 ° range of scatter angles
Filtering processing, finally obtains scattered light intensity distribution curve corresponding to the suspension of the partial size polystyrene spheres.It can disappear in this way
Except scattered light intensity distortion error caused by a variety of causes in measurement, preferably ensure the scattering light of resulting polystyrene spheres suspension
The measurement accuracy of strength distribution curve.
2) using the cell sample of G1 phase, S phase, G2 phase and M phase, the correspondence G1 with same prescribed concentration is respectively prepared
Phase, the S phase, G2 phase and M phase cell suspension and be packed into teat glass, after shaking up respectively, under identical measuring condition, using wide
Angle light scattering apparatus measures the cell suspension of each cell cycle within the scope of 0 ~ 180 ° at interval of the scattered light intensity on 5 ° respectively,
According to measurement result, the corresponding scattered light intensity distribution curve of cell suspension of G1 phase, S phase, G2 phase and M phase are obtained.
In the scattered light intensity distribution curve for specifically measuring 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 within the scope of 0 ~ 180 °
Absolute value be greater than 10% scattered light intensity angle value, then the resulting scattered light intensity angle value of measurements other time on the scattering angle are asked
It is average, scattered light intensity average value of the cell suspension of the cell cycle on corresponding firing angle degree is obtained, 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
It is bent to finally obtain the distribution of scattered light intensity corresponding to the cell suspension of the cell cycle according to fitting and the disposal of gentle filter for line number
Line.Scattered light intensity distortion error caused by a variety of causes in measurement can be eliminated in this way, preferably ensure resulting cell suspension
Scattered light intensity distribution curve measurement accuracy.
3) the corresponding scattered light intensity distribution curve of the cell suspension of G1 phase, S phase, G2 phase and M phase is obtained with preparatory
To the scattered light intensity distribution curves of suspension of variant partial size polystyrene spheres compare, determine respectively the G1 phase, the S phase,
The respective matched polystyrene spheres suspension of institute of the scattered light intensity distribution curve of the cell suspension of G2 phase and M phase, thus will matching
The suspension of obtained each partial size polystyrene spheres is as cell cycle scattered light intensity model.
It can see by above-mentioned process, the present invention utilizes the cell cycle scattered light intensity model foundation side of polystyrene spheres
Method can be established to obtain to be scattered as to the living body biological cell cycle using the suspension of different-grain diameter polystyrene spheres
The cell cycle scattered light intensity model of light-intensity test reference standard, and the property of polystyrene spheres is stablized, and will not generate because thin
Born of the same parents' merisis and cause the cell cycle constantly to change, therefore persistence can be able to, solve permanent guarantor in the prior art
The cell cycle scattered light intensity detection reference standard deposited is difficult to the problem of making.
In the specific implementation, the prescribed concentration of the suspension of made polystyrene spheres can be 106A polystyrene spheres/
mL、2×105A polystyrene spheres/mL, 105A polystyrene spheres/mL or 5 × 104A these four concentration of polystyrene spheres/mL,
The prescribed concentration of made cell suspension can also correspondingly be 106A cell/mL, 2 × 105A cell/mL, 105It is a thin
Born of the same parents/mL or 5 × 104A these four concentration of cell/mL, because the polystyrene spheres suspension and cell of this four concentration ranges are outstanding
Liquid, multiple scattering will not impact scattered light intensity, and if the too low scattered light intensity that will lead to of concentration is very weak, be unfavorable for surveying
Amount, therefore select these four concentration 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 105A polystyrene spheres/mL, 105It is a thin
Born of the same parents/mL, benefit are that the scattered light intensity under the concentration is moderate, can conveniently observe and scatter light between different samples
Strong difference.It, 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, it can be using trypsin solution as reagent dilutions cell sample
To prescribed concentration.And when being scattered luminous intensity measurement using wide-angle light scattering apparatus, preferably with laser as light source,
Advantage is good beam quality, has 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 best, and the performance of dynamic light scattering and measurement knot
Fruit is more preferable at that wavelength.
Illustrate the method for the present invention with embodiment below.It should be understood that these embodiments are only used for into one
Step illustrates embodiment of the present invention, rather than for limiting the present invention.
Embodiment:
Using method of the invention, the cell cycle scattered light intensity model of HeLa cell is established, specifically include the following steps:
1) pre-production prescribed concentration is 105The suspension and dress of the different-grain diameter polystyrene spheres of a polystyrene spheres/mL
Enter in teat glass, after shaking up respectively, using wide-angle light scattering apparatus measure respectively the suspensions of every kind of partial size polystyrene spheres 0 ~
At interval of the scattered light intensity on 5 ° within the scope of 180 °, using laser as light source, and the optical maser wavelength of laser is
532nm, according to measurement result, the corresponding scattered light intensity distribution of suspension for obtaining variant partial size polystyrene spheres is bent
Line.
2) using the HeLa cell sample of G1 phase, S phase, G2 phase and M phase, 10 are pressed respectively5The prescribed concentration system of a cell/mL
It at the HeLa cell suspension for corresponding to G1 phase, S phase, G2 phase and M phase and is packed into teat glass, after shaking up respectively, in identical measurement
Under the conditions of, using wide-angle light scattering apparatus measure respectively the HeLa cell suspension of each cell cycle within the scope of 0 ~ 180 ° at interval of
Scattered light intensity on 5 °, using laser as light source, and the optical maser wavelength of laser is that 532nm is obtained according to measurement result
To the corresponding scattered light intensity distribution curve of HeLa cell suspension of G1 phase, S phase, G2 phase and M phase.
In the present embodiment, the obtained G1 phase, S phase, G2 phase and M phase the corresponding scattered light intensity of HeLa cell suspension
It is as shown in Figure 2 to spend distribution curve.
3) by the corresponding scattered light intensity distribution curve of the HeLa cell suspension of G1 phase, S phase, G2 phase and M phase and in advance
The scattered light intensity distribution curve of the suspension of the variant partial size polystyrene spheres first obtained compares, and determines G1 phase, S respectively
The respective matched polystyrene spheres suspension of institute of the scattered light intensity distribution curve of the HeLa cell suspension of phase, G2 phase and M phase, thus
The suspension for each partial size polystyrene spheres that matching is obtained is as the cell cycle scattered light intensity model of HeLa cell.
It in the present embodiment, is learnt by comparison, the HeLa cell suspension (G1 phase cell) of G1 phase and 12 μm of partial sizes are poly-
The scattered light intensity distribution curve of the suspension (12 μm of PS) of styrene ball matches, as shown in Figure 3；The HeLa cell suspension of S phase
The scattered light intensity distribution curve of the suspension (14 μm of PS) of (S phase cell) and 14 μm of partial size polystyrene spheres matches,
As shown in Figure 4；(11 μm of suspension of the HeLa cell suspension (G2 phase cell) of G2 phase and 11 μm of partial size polystyrene spheres
PS scattered light intensity distribution curve) matches, as shown in Figure 5；And the HeLa cell suspension (M phase cell) of M phase and 22
The scattered light intensity distribution curve of the suspension (22 μm of PS) of μm partial size polystyrene spheres matches, as shown in Figure 6.Just divide as a result,
Not using the suspension of 12 μm, 14 μm, 11 μm and 22 μm partial size polystyrene spheres as HeLa cell in G1 phase, S phase, G2 phase and M phase
Cell cycle scattered light intensity model.
In above-mentioned Fig. 2 ~ Fig. 6, abscissa indicates scatter angle values (angle), and ordinate indicates scattered light intensity
Angle value (light scattering).
In conclusion the present invention utilizes the cell cycle scattered light intensity method for establishing model of polystyrene spheres, can utilize
The suspension of different-grain diameter polystyrene spheres is established to obtain to be scattered light-intensity test ginseng as to the living body biological cell cycle
The cell cycle scattered light intensity model of sighting target standard, and the property of polystyrene spheres is stablized, and will not generate because of cell division growth
And cause the cell cycle constantly to change, therefore persistence can be able to, solve the cell week of persistence in the prior art
Phase scattered light intensity detection reference standard is difficult to the problem of making；Also, since different-grain diameter polystyrene microsphere is diluted to difference
When the concentration of gradient, show angle rely on light scattering characteristic be consistent from the light scattering characteristic of phases different in the cell cycle or
Meet after amendment, therefore there is good universality using the cell cycle scattered light intensity model that the method for the present invention is established,
More cell cycle light intensity of light scattering models will be improved for future and provide expansible thinking, and there is good technical application
Prospect.
Finally, it is stated that the above examples are only used to illustrate the technical scheme of the present invention and are not limiting, although referring to compared with
Good embodiment describes the invention in detail, those skilled in the art should understand that, it can be to skill of the invention
Art scheme is modified or replaced equivalently, and without departing from the objective and range of technical solution of the present invention, should all be covered at this
In the scope of the claims of invention.
Claims (5)
1. a kind of cell cycle scattered light intensity method for establishing model using polystyrene spheres, which is characterized in that pre-production tool
There is the suspension of the different-grain diameter polystyrene spheres of prescribed concentration, be scattered luminous intensity measurement respectively using wide-angle light scattering apparatus,
Obtain the corresponding scattered light intensity distribution curve of suspension of variant partial size 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 period, in identical survey
It is scattered luminous intensity measurement respectively using wide-angle light scattering apparatus under the conditions of amount, obtains the cell suspension of each cell cycle respectively
Corresponding scattered light intensity distribution curve, and the scattered light intensity with the suspension for the variant partial size polystyrene spheres being previously obtained
Distribution curve compares, and determines the scattered light intensity distribution curve of the cell suspension of each different cell cycle respectively institute respectively
Matched polystyrene spheres suspension, so that the suspension for each partial size polystyrene spheres that matching is obtained scatters light as the cell cycle
Strong model；Specifically include the following steps:
1) pre-production has the suspension of the different-grain diameter polystyrene spheres of prescribed concentration and is packed into teat glass, shakes up respectively
Afterwards, the suspensions of every kind of partial size polystyrene spheres is measured respectively using wide-angle light scattering apparatus within the scope of 0 ~ 180 ° at interval of on 5 °
Scattered light intensity obtains the corresponding scattered light intensity of suspension point of variant partial size polystyrene spheres according to measurement result
Cloth curve；In the step, when measuring the scattered light intensity distribution curve of suspension of every kind of partial size polystyrene spheres, duplicate measurements
The suspension of 5 ~ 10 corresponding partial size polystyrene spheres within the scope of 0 ~ 180 ° at interval of the scattered light intensity on 5 °, reject wherein from
The absolute value of equal rate is greater than 10% scattered light intensity angle value, then measures resulting scattered light intensity for other times on the scattering angle
Angle value is averaging, and obtains scattered light intensity average value of the suspension of the partial size polystyrene spheres on corresponding firing angle degree, thus point
The suspension for not obtaining the partial size polystyrene spheres is average at interval of the scattered light intensity on 5 ° in 0 ° ~ 180 ° range of scatter angles
Value, and curve data fit and the disposal of gentle filter are carried out, obtain scattering light corresponding to the suspension of the partial size polystyrene spheres
Strength distribution curve；
2) using the cell sample of G1 phase, S phase, G2 phase and M phase, correspondence G1 phase with same prescribed concentration, S is respectively prepared
Phase, G2 phase and M phase cell suspension and be packed into teat glass, after shaking up respectively, under identical measuring condition, utilize wide-angle light
Scatterometer measures the cell suspension of each cell cycle within the scope of 0 ~ 180 ° at interval of the scattered light intensity on 5 ° respectively, according to
Measurement result obtains the corresponding scattered light intensity distribution curve of cell suspension of G1 phase, S phase, G2 phase and M phase；
3) by the cell suspension of G1 phase, S phase, G2 phase and M phase corresponding scattered light intensity distribution curve be previously obtained
The scattered light intensity distribution curve of the suspension of variant partial size polystyrene spheres compares, and determines G1 phase, S phase, G2 phase respectively
With the respective matched polystyrene spheres suspension of institute of the scattered light intensity distribution curve of the cell suspension of M phase, so that matching be obtained
Each partial size polystyrene spheres suspension as cell cycle scattered light intensity model.
2. utilizing the cell cycle scattered light intensity method for establishing model of polystyrene spheres according to claim 1, feature exists
In, in the step 2, 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 within the scope of 0 ~ 180 °
Absolute value be greater than 10% scattered light intensity angle value, then the resulting scattered light intensity angle value of measurements other time on the scattering angle are asked
It is average, scattered light intensity average value of the cell suspension of the cell cycle on corresponding firing angle degree is obtained, 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
It is bent to finally obtain the distribution of scattered light intensity corresponding to the cell suspension of the cell cycle according to fitting and the disposal of gentle filter for line number
Line.
3. utilizing the cell cycle scattered light intensity method for establishing model of polystyrene spheres according to claim 1, feature exists
In the prescribed concentration of the suspension of made polystyrene spheres is 106A polystyrene spheres/mL, 2 × 105A polystyrene spheres/
mL、105A polystyrene spheres/mL or 5 × 104A polystyrene spheres/mL, the prescribed concentration of made cell suspension are corresponding
It is 106A cell/mL, 2 × 105A cell/mL, 105A cell/mL or 5 × 104A cell/mL.
4. utilizing the cell cycle scattered light intensity method for establishing model of polystyrene spheres according to claim 1, feature exists
In when being scattered luminous intensity measurement using wide-angle light scattering apparatus, using laser as light source.
5. utilizing the cell cycle scattered light intensity method for establishing model of polystyrene spheres according to claim 4, feature exists
In the optical maser wavelength of the laser is 532nm.
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