CN108918898A - The method and its application that vitro detection lipopolysaccharides influences red cell deformability - Google Patents
The method and its application that vitro detection lipopolysaccharides influences red cell deformability Download PDFInfo
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Abstract
The invention discloses a kind of method and its application influenced based on micro-fluidic chip vitro detection lipopolysaccharides on red cell deformability.The present invention is based on micro-fluidic chips can realize individuation detection influence of the lipopolysaccharides to red cell deformability and the screening for improving red cell deformability original new drug；Lipopolysaccharides influences as clinical antibiotics treatment septicopyemia Drug utilization index parameter and curative effect of medication detection application red cell deformability individual；Present invention discover that catalase content, lipid peroxide contents, superoxide dismutase activity, Na+‑K+This six main effective biochemical indicators of-ATPase activity, activity of glutathione peroxidase, amount of ATP play an important role to the change of erythrocyte deformability after lipopolysaccharide.
Description
Technical field
The present invention relates to a kind of method and its application that vitro detection lipopolysaccharides influences red cell deformability.
Background technique
Lipopolysaccharides (LPS) is endotoxic main chemical compositions, it is present in the cell wall of gram negative bacterial cell
In.Cell wall, which is disintegrated, after bacterial death releases endotoxin.When pathogenic bacteria invade blood circulation system, the generations such as toxin can be generated
It thanks to product, causes whole body acute infection, generate septicopyemia.Pyemia is one of clinical critical illness, and the death rate is high, tool
Have and feel cold, generates heat, the characteristics of fash, arthralgia, digestive discomfort and hepatosplenomegaly.Serious case is often with lactic acidosis
With intractable septic shock or migration lesion, then rapid progression is multiple organ failure, eventually leads to death.With
Research is goed deep into, it has been found that there are systemic haemodynamics and extensive microcirculation disorders for sepsis patient.Due to blood stream
Become the hypersensitivity learned, is of great significance for the improving blood flow and monitoring of sepsis patient.Human red cell (RBCs) is made
For host cell most in blood, in hemorheology, microcirculatory perfusion is risen emphatically in O_2 transport and general metabolism stable state
It acts on.Red blood cell shows significant and special deformability, because about 8 microns of its diameter substantially exceed about 3 microns micro-
Capillary, it, which has to pass through deformation, could pass through to guarantee effective operation of microcirculation.Some reports show that red blood cell becomes
Exist between shape and septicemia and be associated with, and finds that the structure of red blood cell and chemical property change in septicopyemia.Face
Septicopyemia is treated on bed and uses antibiotic mostly, and antibiotic can induce bacterium and disintegrate release endotoxin, start antibiotic treatment
Effect of the endotoxin burst size in pyemia and infectious shock pathogenesis is great afterwards.The sensitivity of Different Individual induced by endotoxin
Sex differernce may generate instant adverse effect to patient to influence the effect for the treatment of.
Up to the present, the method for measuring RBCs morphotropism mainly includes micro-pore-film filtration, laser diffractometry, sucking of a tiny amount of milk
Method, atomic force microscope, optical tweezers etc..Wherein, micro-pore-film filtration and laser diffractometry only provide red blood cell population morphotropism
Can, rather than the deformation parameter of individual cells.Micropipette aspiration method and atomic force microscope be exclusively used in measurement individual cells, however its
Measurement is limited by handling capacity.Moreover, this technology has expensive equipment, time-consuming, labour and technology-intensive disadvantage.It is another
Leucocyte (WBCs) deformability parameter of aspect, septic patient also changes, it is known that for measuring the tradition of RBC morphotropism
There are the interference of WBC, RBC washing procedure and the removal pale brown layer operations of blood plasma to be not enough to completely remove leucocyte for technology, if dress
It sets and is blocked by leucocyte, then observed deformable index reduces, and causes to be mistakenly considered erythrocyte deformability reduction.
Although existing research proposes effect of the LPS in vitro to RBC morphotropism, the deep discussion LPS of system is to external
The morphotropism individuation difference and reason of red blood cell, and by by the mechanical property of red blood cell and biochemical index parameter,
Including Na+-K+- ATPase activity, superoxide dismutase activity, activity of glutathione peroxidase, hydrogen peroxide enzyme activity
Property, amount of ATP, lipid peroxide contents, content of hemoglobin and memebrane protein framework ingredient content are divided
Analysis, the research for finding the biochemical indicator mainly influenced are not reported also, are discussed the variation of LPS induction red cell deformability, are taken off
The reason of having shown variation and its clinical meaning.However, as far as we know, sensibility of the red blood cell of Different Individual to LPS so far
The reason of difference is not yet taken off, and the individual of the external evoked red cell deformability of LPS relies on effect is unclear.This technology is wished
By combining microfluidic chip technology with both Biochemical Indexes, important index parameter is found out.By to different
Body analyzes the sensitivity differences of LPS, provides theoretical branch with the personalized medicine of antibiotic treatment septicopyemia for clinic
It holds, while also providing new screening technique to improve the original new drug research and development of red cell deformability.
Summary of the invention
To solve the above problems, the application is by by both microfluidic chip technology and Biochemical Indexes phase
In conjunction with finding out important index parameter, provide new index and theories integration for individualized clinical treatment, and is red thin to improve
The original new drug research and development of born of the same parents' deformability provide new screening technique.
The purpose of the present invention is to provide one kind based on micro-fluidic chip vitro detection lipopolysaccharides to red cell deformability
The method and its application of influence.
The technical solution adopted by the present invention is that：
A method of detection chemical substance influences red cell deformability, by the red blood cell after chemical substance treatment
Sample is added in micro-fluidic chip, detects the rate travel of red blood cell, is compared with the control group without chemical substance treatment
Compared with after chemical substance treatment, whether red cell deformability changes for judgement, to judge that chemical substance becomes red blood cell
The influence of shape ability；This method is not used in the diagnosing and treating of disease.
Further, several injection ports and outlet are contained in micro-fluidic chip, is connected between injection port and outlet
Microchannel；There is retention device at microchannel entrance, prevent non-red blood cell from entering and blocks microchannel, redoubling of attaching most importance in microchannel column
The triangular prism structure of arrangement is divided into 2.8~3.2 μm between triangular prism structure, only a RBC deformation is allowed to pass through；Trigone in microchannel
The columns of rod structure is 10 column or more.
Further, the retention device is attached most importance to the cylindrical structure of multiple arranged in parallel, and 8~12 μm of cylindrical structure diameter, phase
Adjacent cylindrical structure gap is 8~12 μm.
Further, the diameter of injection port and outlet is 1.2~1.7mm.
Further, red blood cell sample is added to before micro-fluidic chip, to red blood cell carry out fluorescent staining, dyeing it is red
After cell is added to micro-fluidic chip, the shift image of red blood cell can be captured according to fluorescence, to measure the movement of red blood cell
Rate.
Further, chemical substance is lipopolysaccharides.
Further, will through lipopolysaccharides, treated that red blood cell sample is added in micro-fluidic chip, detect red blood cell
Rate travel is compared with without the control group that lipopolysaccharides is handled, and is determined after lipopolysaccharides is handled, red cell deformability is
It is no to change, to judge influence of the lipopolysaccharides to red cell deformability；This method is not used in the diagnosing and treating of disease.
The above method is improving the application in the screening of red cell deformability original new drug.
Application of the above method in the drug effect of the antibiotic of detection treatment septicopyemia.
Detect catalase content, lipid peroxide contents, superoxide dismutase activity, Na+-K+-ATPase
The reagent of activity, activity of glutathione peroxidase or/and amount of ATP assists detection lipopolysaccharides to red in preparation
Application in the kit of the influence of cytomorphosis ability.
The beneficial effects of the invention are as follows：
The micro-fluidic chip method that detection lipopolysaccharides influences red cell deformability, energy are given the present invention provides a kind of
Enough dynamic monitoring erythrocyte deformabilities, while unicellular and group's red blood cell deformability parameter index being provided, exclude leucocyte
Interference.The advantages that the application method has, high throughput few to sample consumption, and good portability visualizes, easy to operate, not only
It can provide whole deformability information, and single celled morphotropism information can be provided.
The present invention has found that the main reason for LPS influences erythrocyte deformability is by the measurement to red blood cell biochemical indicator
Oxidative damage factor, and find that red blood cell biochemical indicator includes catalase, lipid peroxide contents, superoxide dismutase
Enzymatic activity, Na+-K+- ATPase, activity of glutathione peroxidase, amount of ATP and erythrocyte deformability have compared with
Strong correlation, related coefficient are followed successively by 0.805, and -0.787,0.626,0.531,0.509 and 0.439.
Present invention discover that LPS is to different healthy volunteer's erythrocyte deformabilities, there are very strong individuation differences.
The present invention can provide new theories integration not only for individualized clinical treatment septicopyemia, but also red thin to improve
The original new drug research and development of born of the same parents' deformability provide new screening technique.Influence due to LPS to erythrocyte deformability has very strong
Individual dependence, therefore for patients with sepsis antibiotic treatment during, the screening based on morphotropism can be achieved
The potential clinical benefit of propertyization medical treatment.
Detailed description of the invention
Fig. 1 is the experimental principle figure of micro-fluidic chip, and A is the step schematic illustration for making chip, and B is that micro-fluidic chip is real
Body figure, C are the microcosmic schematic diagram of micro-fluidic chip, and D is the deformation pattern that the red blood cell of fluorescent staining passes through in microchannel.
Fig. 2 is research of the various concentration LPS to whole blood red cell deformability individual difference.A-I respectively represents different
Blood sample.P value represents medicine group and control group, and (whether LPS concentration red blood cell migration rate between 0pg/mL) has significant difference
It is different：* P is represented<0.05, * * * represents P<0.001.
Fig. 3 is influence of the various concentration LPS to all each biochemical indicators of healthy volunteer's red blood cell, the migration of (A) red blood cell
Speed also refers to morphotropism, (B) superoxide dismutase activity, (C) Na+-K+Atpase activity, (D) amount of ATP,
(E) activity of glutathione peroxidase, (F) catalase activity, (G) lipid peroxide contents and (H) hemoglobin
Content, P refer between medicine group and control group (LPS concentration is 0pg/mL) whether there is significant difference：* P is represented<0.05, * * * refers to
P<0.001, red " * " representative is remarkably decreased, and green " * " represents significant increase.
Fig. 4 is the histogram of membrane skeleton protein content variation after various concentration LPS effect.
Fig. 5 is the red blood cell Na after various concentration LPS effect+-K+The line chart of-ATPase activity change, A-I generation respectively
The different blood sample of table.
Fig. 6 is the histogram of the red blood cell sample amount of ATP variation after various concentration LPS effect, A-I difference
Represent different blood samples.
Fig. 7 is the line chart of the red blood cell sample lipid peroxide contents variation after various concentration LPS effect, and A-I divides
Different blood samples is not represented.
Fig. 8 is the histogram of the red blood cell sample superoxide dismutase activity variation after various concentration LPS effect, A-I
Respectively represent different blood samples.
Fig. 9 is the line chart of the red blood cell sample activity of glutathione peroxidase variation after various concentration LPS effect,
A-I respectively represents different blood samples.
Figure 10 is the line chart of the red blood cell sample catalase Enzyme activities after various concentration LPS effect, and A-I divides
Different blood samples is not represented.
Figure 11 is the histogram of the red blood cell sample content of hemoglobin variation after various concentration LPS effect, A-I generation respectively
The different blood sample of table.
Specific embodiment
The present invention will be further described combined with specific embodiments below.
Embodiment 1
The production of micro-fluidic chip
The production of micro-fluidic chip：Being prepared on silicon wafer using SU-8 photoresist and standardised wet methods etching technics has micro- lead to
After the template in road, PDMS micro-channel chip is prepared using method of molding, main preparation method includes the following steps：
(1) it is placed in vacuum pump with 25mL trim,ethylchlorosilane solution and silicon wafer template, silanization anti-adherency in 1 hour,
The PDMS glue that makes to tear is easier.
(2) the silicon wafer template isopropanol after silanization and ultrapure water clean repeatedly, then are dried with nitrogen and dry.
(3) taking weight ratio is 10：1 PDMS and curing agent, mixes well, and is put into vacuum desiccator vacuum outgas to gas
It after bubble completely disappears, is slowly cast on silicon wafer, avoids generating bubble.95 DEG C solidify 3 hours, then shell PDMS from silicon wafer
From the micro-structure on such silicon wafer has been transferred on flexible PDMS.
(4) in injection port and sample outlet position punching (diameter 1.5mm), then PDMS chip and clean glass slide are put into
After carrying out surface modification treatment in plasma cleaner, PDMS chip is irreversibly adhered on glass slide.
1A is the step schematic illustration for making chip.1B is micro-fluidic chip sterogram, and there are five parallel on a chip
Microchannel, can with 5 experimental groups simultaneously measured on a single die to reduce the individual error of chip.1C is micro-fluidic
The microcosmic schematic diagram of chip.10 μm of the cylindrical pillars diameter of inlet, a height of 4.2 μm, gap be 10 μm, can prevent white blood cell into
Enter and block microchannel, can directly measure whole blood, in channel for triangular prism structure repeat arrangement arranged side by side allow it is high-throughput, three
It is divided into 3 μm between prism structure, only RBC deformation is allowed to pass through.When erythrocyte deformability reduces, speed that red blood cell passes through
Degree will slow down even blocking channel.1D is the deformation pattern that passes through in microchannel of red blood cell of fluorescent staining, and when 0s shows red thin
Born of the same parents just start deformation by the gap between triangular prism structure, most of volume of red blood cell also on the left of triangular prism structure gap,
Do not pass through gap；Indicate that red blood cell is passing through the gap between triangular prism structure when 0.2s and 0.4s, big portion's volume has passed through three
Prism structure gap, small part volume is also on the left of gap；Indicated when 0.6s red blood cell all by triangular prism structure gap,
It is prepared to enter into next triangular prism structure gap.
Embodiment 2
Detection method
1. the preparation of blood sample
Being configured to final concentration with physiological saline is respectively 0ng/mL, 1ng/mL, 10ng/mL, the LPS sample of 100ng/mL.
Blood sample is collected from healthy volunteer's venous puncture, fresh whole blood is divided into four parts, 0ng/mL, 1ng/ is added
The LPS sample of mL, 10ng/mL, 100ng/mL make final LPS concentration be respectively 0pg/mL, 20pg/mL, 200pg/mL,
2000pg/mL, 37 DEG C are incubated for 3 hours.After the completion of incubation, 300g is centrifuged 5 minutes and separates and collects upper plasma and lower layer's red blood cell.
Upper plasma is placed at -20 DEG C and saves backup, and to remove excessive LPS, then exists for lower layer's red blood cell brine 3 times
It is saved backup at 4 DEG C.
2. the measurement of erythrocyte deformability
Before measuring erythrocyte deformability, 1640 culture medium of RPMI containing 20%FBS solution is injected into microchannel
In, it is incubated at room temperature 20 minutes and PDMS wall is modified with coating, prevent its non-specific adhesion.Various concentration LPS is taken to be incubated for
10 μ L of red blood cell, the Cell Tracker Orange fluorescent dye with final concentration of 1 μM mix, and 37 DEG C of incubation 15min are to red thin
Born of the same parents dye.300g centrifuge washing removes excess dye three times after dyeing.It will be red with the RPMI culture medium containing 20%FBS
Cell dilutes to obtain the red blood cell sample of 1% hematocrit.The blood sample of about 3 μ L is taken to be loaded into the sample introduction reservoir of microfluidic device
In, it is vertical fixed by one, the syringe that the inside is partially filled with the 60mL of the RPMI culture medium of 20%FBS buffer be connected into
Mouthful reservoir, to obtain 0.36Pa/ μm of constant pressure.Micro flow control chip device is mounted on equipped with CCD camera
On Leica fluorescence inverted microscope, to capture the shift image of red blood cell.The every 250ms of LAX S software obtains fluorogram automatically
Picture, and post analysis is carried out using metlab and image J software.Red cell deformability is defined as rate travel, refers to list
The shift length of a red blood cell is divided by displacement time.In order to preferably illustrate the variation of erythrocyte deformability speed caused by LPS,
Using " normalizated velocity ".The definition of normalizated velocity is the speed by red blood cell single under various experiment conditions divided by control group
The average speed of whole red blood cell.Fig. 3 is shown in influence of the LPS to healthy volunteer's whole blood erythrocyte deformability, shows different
Body is different to the sensibility of LPS, to be shown as the otherness of the morphotropism of RBC.
3. red blood cell biochemical parameter measures
Each group whole blood blood sample after being incubated for LPS carries out red blood cell biochemical analysis simultaneously.Respectively to red blood cell Na+-K+-
ATPase activity, superoxide dismutase activity, activity of glutathione peroxidase, catalase activity, Adenosine triphosphate
Glycosides content, lipid peroxide contents, the measurement of content of hemoglobin and memebrane protein framework ingredient content are built up according to Nanjing
Each indicator reagent box of Bioengineering Research Institute's purchase carries out time-and-motion study.
4. statistical analysis
Further to inquire into LPS to the influencing mechanism of red cell deformability, biochemical marker data is summarized in one by us
It rises, correlation analysis and one-way analysis of variance is carried out to each biochemical indicator and erythrocyte deformability, obtain Fig. 4.For the purposes of
Diversity of individuals is presented, LPS shows the effect of all subjects in Fig. 5~11 respectively.
It tests all data and is represented as mean+SD.With 17.0 software of SPSS, using single factor test variance point
The statistical significance of analysis and minimum significant sex differernce check analysis each experimental group and control group.P value is less than 0.05 for significant
Sex differernce.
As a result
Result of study of the various concentration LPS to whole blood red cell deformability individual difference
Various concentration LPS is shown in Fig. 2 to the result of study of whole blood red cell deformability individual difference, from Fig. 2's
The result shows that LPS has strong individual difference in erythrocyte deformability.When LPS dosage is that 20pg/mL is handled, observe complete
The movement speed of blood sample A, E and I significantly reduces (Fig. 2, sample E and I, P<0.05；Sample A, P<0.001, LSD examines), with
The increase of LPS concentration, the migration velocity of sample A, E and I are also lower and lower, show the significant decrease of erythrocyte deformability.This
As a result illustrate subject A, the sensibility of E and I induced by endotoxin is high.There are 3 samples to give and is giving 2000pg/mL LPS processing
When, compared with the control group, RBC morphotropism just significantly decreases figure (Fig. 2, B and F, P<0.05；D, P<0.001, LSD inspection
Test), show that the LPS processing of low concentration is less intense to the red blood cell toxic action of these three whole blood samples.Sample C, G and H
Under the LPS effect of 200pg/mL, erythrocyte deformability significantly declines (Fig. 2, G and H, P<0.05；C, P<0.001, LSD inspection
It tests).All samples all show that red blood cell significantly hardens, morphotropism is more significant at highest LPS concentration 2000pg/mL
It reduces, this may be because the damage that generates to sample erythrocyte deformability of high concentration LPS is big.This is the result shows that RBC deformation
Property dose-dependant otherness reaction may be related with sensibility difference of the individual to LPS.LPS is to the bright of red cell deformability
Aobvious individual difference implies that drug individualized treatment should arouse attention, clinically common antibiotics treatment septicopyemia, and antibiosis
Element can destroy bacteria cell wall and discharge endotoxin, should formulate different therapeutic strategies for the patient of induced by endotoxin sensitivity,
To improve the drug compliance of patient.
Influence of the various concentration LPS to each biochemical indicator of Different Individual red blood cell
Fig. 3 is influence of the various concentration LPS to all each biochemical indicators of healthy volunteer's red blood cell.Fig. 3 A display is passed through
Whole blood erythrocyte deformability after LPS is incubated for significantly reduces.LPO is the product of body radical damage.With oxidation level
Increase, response to oxidative stress enhancing, LPO content increases, therefore LPO is able to reflect internal oxygen radical to a certain extent
How much.SOD is intracorporal radicals scavenging enzyme, can react the oxidant defense and protective response ability of body.Due to the work of LPS
With cell generates the substances such as excessive oxygen radical, and SOD consumption increases and content activity declines.As shown in Fig. 3 B and Fig. 3 G, from
Figure we have found that with LPS concentration raising, LPO content increase, while SOD activity it is also on a declining curve.And LPO exists
It is significantly increased when 200pg/mL, and SOD enzyme activity is just significantly reduced in 2000pg/mL, it may be possible to low concentration 20pg/mL's
It after LPS effect, increases to SOD enzyme activity elder generation compensatory, but when LPS induction generates free radicals substantial increase, cell loses generation
It repays, so that cell membrane damage makes the reduction of its enzymatic activity.This is the result shows that the reduction of cellular anti-oxidant defence capability and red blood cell become
Shape reduces related.Patients with sepsis is clinically treated, mitigates the peroxidatic reaction of lipid of LPS induction, is beneficial to disease
Recovery and lapse to.
When endotoxemia occurs, endotoxin can induce body and generate free radicals attack zymophore, thus zymoprotein
Structure changes and loses former activity.This experiment discovery, after LPS acts on whole blood red blood cell, as shown in Figure 3 C, Na+-K+- ATPase is living
Property is in be remarkably decreased.Work as Na+-K+When-ATPase activity declines, Na in red blood cell can lead to+Retention and K+It is lost, causes thin
Born of the same parents' oedema, so that deformability reduces.
RBC special deformability is mainly attributed to its unique bi-concave shape structure and film-cytoskeleton phase interaction
With the two is considered to the height adjustment by ATP.ATP is the main matter of supplying energy.When pyemia microcirculation disorder,
Glycolysis is obstructed, ATP intracellular largely consume content reduction, red blood cell ATP exhaustion will lead to its shape have it is discoid to spine shape ball
The transformation of shape red blood cell also dramatically increases film bending stiffness after ATP consumption.In addition ATP is reduced, and film calcium energy supply is deficient, altogether
It is same that red cell deformability is caused to decline.As shown in Figure 3D, under 2000pg/mL high concentration LPS effect, under ATP content is significant
Drop.
GSH-PX and CAT is enzyme important in erythrocyte antioxidant system.SOD is catalyzed ultra-oxygen anion free radical disproportionation life
At hydrogen peroxide and oxygen molecule.H of the GSH-PX in addition to decomposing low concentration2O2, also there is the energy for the Primary product ROOH for understanding LPO
Power, to inhibit the lipid peroxidation of erythrocyte membrane.Work as H2O2When substantial increase, CAT is responsible for being broken down into water and oxygen,
Activity can reflect that the oxidation resistance of body, experimental result as shown in figures 3 e and 3f, find the two enzymatic activity to a certain extent
It is all substantially reduced compared with the control group, illustrates that red blood cell lipid peroxidation capacity after LPS is acted on enhances, under antioxidase function
Drop.As shown in Fig. 3 H and Fig. 4, variation is not obvious for the variation of red blood cell HB content and memebrane protein after LPS effect.
In order to probe into 9 healthy volunteer's red blood cells significantly reduced mechanism of deformability after LPS is incubated for, we are investigated
Red blood cell biochemical index and the correlation between morphotropism, as a result as shown in Fig. 5~11, as the result is shown catalase,
Lipid peroxide contents, superoxide dismutase activity, Na+-K+- ATPase, activity of glutathione peroxidase, three phosphorus
Adenosine monophosphate content has preferable correlation with erythrocyte deformability, and related coefficient is followed successively by 0.805, and -0.787,0.626,
0.531,0.509 and 0.439.Content of hemoglobin and memebrane protein framework ingredient changes of contents are unobvious.Based on the above results,
It is considered that its oxidative damage is the key factor for influencing red cell deformability after LPS effect red blood cell.
Claims (10)
1. a kind of method that detection chemical substance influences red cell deformability, it is characterised in that：It will be through chemical substance treatment
Red blood cell sample afterwards is added in micro-fluidic chip, detects the rate travel of red blood cell, with pair without chemical substance treatment
It is compared, determines after chemical substance treatment, whether red cell deformability changes, to judge chemical substance according to group
Influence to red cell deformability；
The above method is not used in the diagnosing and treating of disease.
2. according to the method described in claim 1, it is characterized in that：Contain several injection ports and out sample in micro-fluidic chip
Mouthful, microchannel is connected between injection port and outlet；There is retention device at microchannel entrance, prevent non-red blood cell from entering and blocks
Microchannel, the triangular prism structure for multiple arranged in parallel of attaching most importance in microchannel are divided into 2.8~3.2 μm between triangular prism structure, only allow red
Cytomorphosis passes through；The columns of triangular prism structure is 10 column or more in microchannel.
3. according to the method described in claim 2, it is characterized in that：The retention device is attached most importance to the cylinder knot of multiple arranged in parallel
Structure, 8~12 μm of cylindrical structure diameter, adjacent column structure interval is 8~12 μm.
4. according to the method described in claim 2, it is characterized in that：The diameter of injection port and outlet is 1.2~1.7mm.
5. according to the method described in claim 1, it is characterized in that：Red blood cell sample is added to before micro-fluidic chip, to red
Cell carries out fluorescent staining, and after the red blood cell of dyeing is added to micro-fluidic chip, the migration of red blood cell can be captured according to fluorescence
Image, to measure the rate travel of red blood cell.
6. described in any item methods according to claim 1~5, which is characterized in that chemical substance is lipopolysaccharides.
7. according to the method described in claim 6, it is characterized in that, will treated that red blood cell sample is added to is micro- through lipopolysaccharides
In fluidic chip, the rate travel of red blood cell is detected, is compared with without the control group that lipopolysaccharides is handled, is determined through lipopolysaccharides
After processing, whether red cell deformability changes, to judge influence of the lipopolysaccharides to red cell deformability；
The above method is not used in the diagnosing and treating of disease.
8. the method according to claim 11 is improving the application in the screening of red cell deformability original new drug.
9. application of the method according to claim 11 in the drug effect of the antibiotic of detection treatment septicopyemia.
10. detecting catalase content, lipid peroxide contents, superoxide dismutase activity, Na+-K+- ATPase is living
Property, activity of glutathione peroxidase or/and amount of ATP reagent assist detection lipopolysaccharides to red thin in preparation
The application in kit that born of the same parents' deformability influences.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112557261A (en) * | 2020-12-07 | 2021-03-26 | 昆明理工大学 | Erythrocyte separation detection device and separation detection method based on C-shaped microcolumn |
| CN114002130A (en) * | 2021-08-25 | 2022-02-01 | 陕西师范大学 | Method and device for measuring cell deformability |
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2018
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Cited By (3)
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| CN112557261A (en) * | 2020-12-07 | 2021-03-26 | 昆明理工大学 | Erythrocyte separation detection device and separation detection method based on C-shaped microcolumn |
| CN112557261B (en) * | 2020-12-07 | 2022-12-09 | 昆明理工大学 | Erythrocyte separation detection device and separation detection method based on C-shaped microcolumn |
| CN114002130A (en) * | 2021-08-25 | 2022-02-01 | 陕西师范大学 | Method and device for measuring cell deformability |
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