CN115873712B - Deep hole cell culture plate, preparation method and application thereof - Google Patents
Deep hole cell culture plate, preparation method and application thereof Download PDFInfo
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Abstract
The invention discloses a deep hole cell culture plate, a preparation method and application thereof, and relates to the technical field of cell culture devices in biology. The deep hole cell culture plate comprises a basic deep hole cell plate and a surface modified compound; the compound is connected to the surface of the deep hole cell plate through a chemical bond; the above-mentioned compounds include hexadienyl tartaric acid diamine and itaconic acid derivatives; the itaconic acid derivatives include products of ethylhydrazine derived itaconic acid; the grafting rate of the compound on the surface of the deep hole cell plate is more than 0.4 percent. The deep hole cell culture plate prepared by the invention has better surface hydrophilicity, further improves hydrophilicity timeliness and obviously prolongs the service life; and simultaneously, the mechanical property of the cell culture plate is further improved, and the toughness is improved.
Description
Technical Field
The invention belongs to the technical field of cell culture devices in biology, and particularly relates to a deep hole cell culture plate, a preparation method and application thereof.
Background
Deep well plates are used for processing, transferring and storing liquid samples, and are currently widely used in biological fields such as cell culture and the like; the method is used for storing laboratory liquids such as polar organic solutions, acidic and alkaline solutions and the like; a master sampling and storage, mechanical sampling and automatic pipetting system for the authentication system; the device is used for an automatic diluter and a sample adder; for blood coagulation reaction, determination of antibody titer. The requirements of various fields on the environment are different, and the deep hole plate is required to have good tolerance, temperature resistance, high mechanical strength and the like. The upper part of a square hole of the deep hole plate structure in the prior art is easy to generate sample residue or capillary phenomenon, so that excessive sample loss or cross infection is caused. Currently, deep hole plates in the market are mainly made of polypropylene materials, and the polypropylene materials have hydrophobic characteristics due to the non-polarity. The most used method for modifying polypropylene is blending and melting modification, and small-molecule hydrophilic additives are blended, so that after thermal processing, hydrophilic small molecules have poor compatibility with polypropylene materials and can migrate to the surfaces of the materials, thereby enhancing the surface hydrophilicity of the polypropylene, but the materials are easy to generate problems of thermal decomposition, yellowing, mechanical property reduction and the like during high-temperature processing. Secondly, the scheme of surface coating modification is adopted, and the method generally adopts an organosilicon coating, such as a silane coupling agent, has the same pollution problem of entering a solution system, has complex process, has process pollution risk, and has low mass production effect and high production cost.
Disclosure of Invention
The invention aims to provide a deep hole cell culture plate, a preparation method and application thereof, wherein the deep hole cell culture plate has better surface hydrophilicity, the hydrophilicity and timeliness are further improved, and the service life is obviously prolonged; and simultaneously, the mechanical property of the cell culture plate is further improved, and the toughness is improved.
The technical scheme adopted by the invention for achieving the purpose is as follows:
a deep-well cell culture plate, comprising a basic deep-well cell plate and a surface-modified compound; the compound is connected to the surface of the deep hole cell plate through a chemical bond; the above-mentioned compounds include hexadienyl tartaric acid diamine and itaconic acid derivatives; the itaconic acid derivatives include products of ethylhydrazine derived itaconic acid; the grafting rate of the compound on the surface of the deep hole cell plate is more than 0.4 percent. According to the invention, hexadienyl tartaric acid diamine and itaconic acid derivatives are adopted to carry out grafting modification on the surface of the deep hole cell plate activated by argon-oxygen mixed low-temperature plasma, so that the hydrophilic performance of the surface of the deep hole cell culture plate can be effectively improved, and the water contact angle of the deep hole cell culture plate is obviously increased; and the hydrophilic timeliness is remarkably improved, and the product still has excellent hydrophilic performance after being placed in the air for two months, so that the service life is effectively prolonged. Meanwhile, after the surface grafting treatment of the deep hole cell plate, the mechanical property of the deep hole cell plate is improved to a certain extent, and particularly the impact strength is obviously improved. The reason for the method is probably that the surface of the deep hole cell plate is firstly subjected to activation treatment by adopting plasma, then the surface of the deep hole cell plate is subjected to grafting modification by adopting hexadienyl tartaric acid diamine and itaconic acid derivatives, more polar functional groups are introduced into the surface of the culture plate, and new physical and chemical properties are endowed to the material, so that the surface of the deep hole cell culture plate shows better hydrophilicity and the biocompatibility is improved; the hexadiene diamine tartrate and the itaconic acid derivative are connected to the surface of the plate through chemical bonds, the binding force is obviously enhanced, and under the condition that the hexadiene diamine tartrate and the itaconic acid derivative exist at the same time, the hexadiene diamine tartrate and the itaconic acid derivative are cooperatively compounded and better adhere to the surface of the culture plate, so that the hydrophilic timeliness of the surface of the culture plate is further prolonged, and meanwhile, the mechanical property is enhanced.
In a specific embodiment, the compound is modified to the surface of a deep well cell plate by a solid phase infiltration grafting method.
In a specific embodiment, the chemical structure of the itaconic acid derivative is shown in formula I:
the invention also discloses a preparation method of the itaconic acid derivative, which comprises the following steps: the itaconic acid derivative is prepared by adopting ethylhydrazine and itaconic anhydride through amidation reaction.
Further, the preparation method of the itaconic acid derivative specifically comprises the following steps:
and (3) adding chloroform into itaconic anhydride for dissolution, heating to 20-30 ℃ in a water bath, slowly adding ethyl hydrazine-chloroform solution with the concentration of 0.5-1 g/mL, reacting at constant temperature for 30-60 min after dripping, cooling for crystallization, vacuum filtering, recrystallizing the product with chloroform, vacuum distilling, and drying to obtain the itaconic acid derivative.
In a specific embodiment, the molar ratio of itaconic anhydride to ethylhydrazine is 1-1.2:1.
The invention also discloses a preparation method of the deep hole cell culture plate, which comprises the following steps:
pretreating, namely placing the deep-hole cell plate in an ultrasonic condition, and cleaning the deep-hole cell plate by sequentially adopting distilled water and acetone to obtain a pretreated deep-hole cell plate;
plasma activation with Ar/O 2 Mixing low-temperature plasmas to perform activation treatment on the surface of the pretreated deep-hole cell plate to obtain an activated deep-hole cell plate;
and (3) grafting, namely soaking the activated deep-hole cell plate in an aqueous solution containing hexadiene diamine tartrate and itaconic acid derivatives for treatment, and grafting to obtain the deep-hole cell culture plate.
Specifically, the preparation method of the deep hole cell culture plate comprises the following steps:
pretreating, namely placing the deep hole cell plate in distilled water under an ultrasonic condition for 20-40 min, performing ultrasonic washing with acetone for 20-40 min, and then performing vacuum drying at 70-80 ℃ for 10-20 min;
activating plasma, placing the pretreated deep hole cell plate into a plasma reaction chamber in a radio frequency plasma system, starting a mechanical pump and a Roots pump in sequence, and waiting for the working air pressure in the reaction chamber to be 10% -3 Setting process parameters in Pa; taking out the sample after the treatment is finished to obtain an activated deep hole cell plate;
and (3) grafting, namely preparing an aqueous solution containing hexadiene diamine tartrate and itaconic acid derivatives in advance, heating to 60-70 ℃, adding the aqueous solution into a deep hole cell plate after plasma activation to submerge the part to be modified of the deep hole cell plate, grafting a compound on the surface of the part to be modified of the deep hole cell plate by adopting a solid-phase infiltration grafting method for 2-4 min, taking out, washing for 2-5 times by using water, and drying to obtain the deep hole cell culture plate.
In particular embodiments, ar/O 2 Mixing low temperature plasma process parameters, comprising: ar/O 2 The flow ratio is 38-40/10-13 sccm, the treatment time is 4-6 min, the power is 280-320W, and the working air pressure is 75-85 Pa.
In a specific embodiment, the concentration of hexadiene diamine tartrate in the aqueous solution is 40-45 wt%.
In a specific embodiment, the concentration of itaconic acid derivatives in the aqueous solution is 20 to 25wt%.
In a specific embodiment, the aqueous solution is replaced by a water/acetone mixed solution.
Preferably, the above water/acetone mixed solution is further added with 3-bis (2-hydroxyethyl) amino-2-hydroxypropanesulfonic acid. According to the invention, 3-bis (2-hydroxyethyl) amino-2-hydroxy propane sulfonic acid is added in the surface grafting treatment process of the deep hole cell plate, so that a certain surfactant effect can be realized, the grafting process of the grafted compound on the surface of the deep hole cell plate is better promoted, the grafting rate of the compound is obviously increased, and further, the morphology structure of the surface of the deep hole cell culture plate is beneficially changed, so that the hydrophilicity and mechanical property of the deep hole cell culture plate are effectively improved.
In a specific embodiment, the addition amount of the 3-bis (2-hydroxyethyl) amino-2-hydroxypropanesulfonic acid is 0.01-0.02 mol/L; the volume ratio of the acetone to the water is 0.8-1.4:1.
More preferably, the compound grafting rate on the surface of the deep well cell plate is > 0.7%.
Compared with the prior art, the invention has the following beneficial effects:
according to the invention, hexadienyl tartaric acid diamine and itaconic acid derivatives are adopted to carry out grafting modification on the surface of the deep hole cell plate activated by argon-oxygen mixed low-temperature plasma, so that the hydrophilic performance of the surface of the deep hole cell culture plate can be effectively improved, and the water contact angle of the deep hole cell culture plate is obviously increased; and the hydrophilic timeliness is obviously improved, and the service life is effectively prolonged. Meanwhile, after the surface grafting treatment of the deep hole cell plate, the mechanical property of the deep hole cell plate is improved to a certain extent, and particularly the impact strength is obviously improved. According to the invention, 3-bis (2-hydroxyethyl) amino-2-hydroxy propane sulfonic acid is added in the surface grafting treatment process of the deep hole cell plate, so that the grafting rate of the compound can be obviously increased, and further, the morphology structure of the surface of the deep hole cell culture plate is beneficially changed, so that the hydrophilicity and mechanical property of the deep hole cell culture plate are effectively improved.
Therefore, the invention provides a deep hole cell culture plate, a preparation method and application thereof, wherein the deep hole cell culture plate has better surface hydrophilicity, the hydrophilicity and timeliness are further improved, and the service life is obviously prolonged; and simultaneously, the mechanical property of the cell culture plate is further improved, and the toughness is improved.
Drawings
FIG. 1 is the infrared test results of the activated deep-well cell plates and deep-well cell culture plates prepared in example 1 of the present invention.
Detailed Description
For the purpose of making the objects, technical solutions and advantages of the embodiments of the present invention more apparent, the following describes in detail various embodiments of the present invention with reference to the embodiments. However, those of ordinary skill in the art will understand that in various embodiments of the present invention, numerous technical details have been set forth in order to provide a better understanding of the present application. However, the technical solutions claimed in the present application can be implemented without these technical details and with various changes and modifications based on the following embodiments.
The deep hole cell plate used in the embodiment of the invention is made of polypropylene.
Example 1:
preparation of deep hole cell culture plate:
pretreating, namely placing the deep hole cell plate in distilled water under ultrasonic conditions for washing for 35min, performing ultrasonic washing with acetone for 35min, and then performing vacuum drying at 76 ℃ for 15min;
activating plasma, placing the pretreated deep hole cell plate into a plasma reaction chamber in a radio frequency plasma system, starting a mechanical pump and a Roots pump in sequence, and waiting for the working air pressure in the reaction chamber to be 10% -3 And setting process parameters during Pa, wherein the process parameters comprise: ar/O 2 The flow ratio is 39/11sccm, the treatment time is 4.5min, the power is 295W, and the working air pressure is 82Pa; taking out the sample after the treatment is finished to obtain a deep hole cell plate after plasma activation;
and (3) grafting, namely preparing an aqueous solution containing hexadiene diamine tartrate and itaconic acid derivatives in advance (wherein the concentration of the hexadiene diamine tartrate is 43wt% and the concentration of the itaconic acid derivatives is 23 wt%), heating to 65 ℃, then adding the aqueous solution into a deep hole cell plate subjected to plasma activation to submerge the part to be modified of the deep hole cell plate, grafting a compound on the surface of the part to be modified of the deep hole cell plate by adopting a solid-phase infiltration grafting method for 3min, taking out, washing for 4 times by using water, and drying to obtain the deep hole cell culture plate.
Preparation of the itaconic acid derivatives:
dissolving itaconic anhydride in chloroform, heating to 28 ℃ in a water bath, slowly adding an ethyl hydrazine-chloroform solution (the molar ratio of itaconic anhydride to ethyl hydrazine is 1.15:1) with the concentration of 0.8g/mL, reacting at constant temperature for 45min after the dripping is finished, cooling for crystallization, performing vacuum suction filtration, recrystallizing the product with chloroform, performing vacuum distillation, and drying to obtain the itaconic acid derivative (the chemical structure is shown as follows). 1 H NMR(500 MHz, D 2 O) δ 6.82、5.93(2H, C=CH 2 ), 2.84 (s, 2H, O=C-CH 2 ), 2.52 (m, 2H, -CH 2 ), 1.09 (t, 3H, -CH 3 )。
Example 2:
the preparation of deep well cell culture plates differs from example 1 in that:
plasma activation process parameters, including: ar/O 2 The flow ratio is 38/10sccm, the treatment time is 4min, the power is 320W, and the working air pressure is 75Pa;
the concentration of hexadienyl diamine tartrate in the aqueous solution was 40wt% and the concentration of itaconic acid derivative was 20wt%.
Itaconic acid derivatives were prepared as in example 1.
Example 3:
the preparation of deep well cell culture plates differs from example 1 in that:
plasma activation process parameters, including: ar/O 2 The flow ratio is 40/12sccm, the treatment time is 5min, the power is 280-320W, and the working air pressure is 85Pa;
the concentration of hexadienyl diamine tartrate in the aqueous solution was 45wt% and the concentration of itaconic acid derivative was 25wt%.
Itaconic acid derivatives were prepared as in example 1.
Example 4:
the preparation of deep well cell culture plates differs from example 1 in that:
plasma activation process parameters, including: ar/O 2 The flow ratio is 40/10sccm, the treatment time is 5.5min, the power is 310W, and the working air pressure is 80Pa;
the concentration of hexadienyl diamine tartrate in the aqueous solution was 44wt% and the concentration of itaconic acid derivative was 22wt%.
Itaconic acid derivatives were prepared as in example 1.
Example 5:
the preparation of deep well cell culture plates differs from example 1 in that:
in the grafting treatment process, the itaconic acid derivative is replaced by hexadiene diamine tartrate with the same molar quantity in the aqueous solution.
Example 6:
preparation of deep hole cell culture plate:
pretreating, namely placing the deep hole cell plate in distilled water under ultrasonic conditions for washing for 35min, performing ultrasonic washing with acetone for 35min, and then performing vacuum drying at 76 ℃ for 15min;
activating plasma, placing the pretreated deep hole cell plate into a plasma reaction chamber in a radio frequency plasma system, starting a mechanical pump and a Roots pump in sequence, and waiting for the working air pressure in the reaction chamber to be 10% -3 And setting process parameters during Pa, wherein the process parameters comprise: ar/O 2 The flow ratio is 39/11sccm, the treatment time is 4.5min, the power is 295W, and the working air pressure is 82Pa; taking out the sample after the treatment is finished to obtain a deep hole cell plate after plasma activation;
and (3) carrying out grafting treatment, namely preparing water/acetone solution containing hexadiene diamine tartrate and itaconic acid derivatives in advance, wherein the concentration of the hexadiene diamine tartrate is 43wt%, the concentration of the itaconic acid derivatives is 23wt%, the volume ratio of acetone to water is 1:1, adding 3-bis (2-hydroxyethyl) amino-2-hydroxy propane sulfonic acid (the adding amount is 0.015 mol/L), heating to 65 ℃, then adding the mixture into a deep hole cell plate subjected to plasma activation to submerge the part to be modified of the deep hole cell plate, grafting a compound on the surface of the part to be modified of the deep hole cell plate by adopting a solid-phase infiltration grafting method, taking out, washing for 4 times by using water, and drying to obtain the deep hole cell culture plate.
Itaconic acid derivatives were prepared as in example 1.
Example 7:
preparation of deep hole cell culture plate:
pretreating, namely placing the deep hole cell plate in distilled water under ultrasonic conditions for washing for 35min, performing ultrasonic washing with acetone for 35min, and then performing vacuum drying at 76 ℃ for 15min;
activating plasma, placing the pretreated deep hole cell plate into a plasma reaction chamber in a radio frequency plasma system, starting a mechanical pump and a Roots pump in sequence, and waiting for the working air pressure in the reaction chamber to be 10% -3 And setting process parameters during Pa, wherein the process parameters comprise: ar/O 2 The flow ratio is 39/11sccm, the treatment time is 4.5min, the power is 295W, and the working air pressure is 82Pa; taking out the sample after the treatment is finished to obtain a deep hole cell plate after plasma activation;
and (3) carrying out grafting treatment, namely preparing a water/acetone solution containing hexadiene diamine tartrate and itaconic acid derivatives in advance, wherein the concentration of the hexadiene diamine tartrate is 43wt%, the concentration of the itaconic acid derivatives is 23wt%, the volume ratio of acetone to water is 1:1, heating to 65 ℃, then adding the mixture into a plasma activated deep hole cell plate to submerge the to-be-modified part of the deep hole cell plate, grafting a compound on the surface of the to-be-modified part of the deep hole cell plate by adopting a solid phase infiltration grafting method for 3min, taking out, washing with water for 4 times, and drying to obtain the deep hole cell culture plate.
Itaconic acid derivatives were prepared as in example 1.
Example 8:
the preparation of deep well cell culture plates differs from example 6 in that:
in the grafting treatment process, the itaconic acid derivative is replaced by hexadiene diamine tartrate with the same molar quantity in the aqueous solution.
Comparative example 1:
preparation of deep hole cell culture plate:
pretreating, namely placing the deep hole cell plate in distilled water under ultrasonic conditions for washing for 35min, performing ultrasonic washing with acetone for 35min, and then performing vacuum drying at 76 ℃ for 15min;
activating plasma, placing the pretreated deep hole cell plate into a plasma reaction chamber in a radio frequency plasma system, starting a mechanical pump and a Roots pump in sequence, and waiting for the working air pressure in the reaction chamber to be 10% -3 And setting process parameters during Pa, wherein the process parameters comprise: ar/O 2 The flow ratio is 39/11sccm, the treatment time is 4.5min, the power is 295W, and the working air pressure is 82Pa; and taking out the sample after the treatment is finished to obtain the deep hole cell plate after the plasma activation.
Test example 1:
infrared sign
The test was performed using a fourier transform infrared spectrometer. Wave number range 4000-500 cm -1 。
The above test was performed on the activated deep-well cell plate prepared in example 1 and the deep-well cell culture plate, and the results are shown in fig. 1. From the analysis in the figure, it can be seen that 3250cm in the IR spectrum of the deep-well cell culture plate compared with the IR test results of the activated deep-well cell culture plate -1 ~3400cm -1 Characteristic absorption peak of N-H bond appears in the range of 1670cm -1 、1538cm -1 The characteristic absorption peaks of the amide groups appear nearby, and the results show that the medium-deep hole cell culture plate of the example 1 is successfully prepared.
Test example 2:
hydrophilic property measurement
And measuring the contact angle, namely placing a sample to be measured on a static contact angle measuring instrument stage, fixing and compacting, then taking 5 mu L of deionized water by a microsyringe, dripping the deionized water on the surface of the sample, photographing the form of the surface water drop after 10s, measuring the contact angle between the liquid drop and the surface of the sample by adopting software treatment, and repeating the test for 5 times in an experiment to obtain an average value.
Time-effectiveness of hydrophilic Properties
The prepared sample was placed in air (25.+ -. 3 ℃ C., 50% humidity) for 60d, where the contact angle of the surface was measured, and the contact angle increase rate was calculated to characterize the hydrophilicity timeliness of the sample.
The deep well cell culture plates prepared in examples 1 to 8 and comparative example 1 were subjected to the above test, and the results are shown in Table 1:
TABLE 1 hydrophilic property test results
| Sample of | Contact angle/° | Contact angle increase rate/% |
| Comparative example 1 | 37 | 200.8 |
| Example 1 | 26 | 2.9 |
| Example 2 | 25 | 2.8 |
| Example 3 | 26 | 3.0 |
| Example 4 | 25 | 2.9 |
| Example 5 | 33 | 20.4 |
| Example 6 | 18 | 2.7 |
| Example 7 | 23 | 2.8 |
| Example 8 | 27 | 19.9 |
As can be seen from the data analysis in Table 1, the surface water contact angle of the deep hole cell plate prepared in example 1 is obviously lower than that of the deep hole cell plate prepared in example 5 and comparative example 1, and after the deep hole cell plate is placed in air for 60 days, the surface water contact angle increase rate is obviously lower than that of the deep hole cell plate prepared in example 5 and comparative example 1, and the effect of example 5 is better than that of comparative example 1, so that the chemical grafting modification of the deep hole cell plate by adopting itaconic acid derivatives can effectively improve the hydrophilic performance of the surface of the deep hole cell culture plate, effectively improve the hydrophilic timeliness of the surface and prolong the service life of the product; meanwhile, under the condition that hexadiene diamine tartrate exists together, the effect of enhancing the hydrophilicity of the surface of the deep hole cell culture plate is better. The deep well cell plate prepared in example 6 has a significantly lower surface water contact angle than that of example 7, the effect of example 7 is slightly better than that of example 1, and the effect of example 8 is better than that of example 5, indicating that the addition of 3-bis (2-hydroxyethyl) amino-2-hydroxypropanesulfonic acid during the grafting treatment can further increase the hydrophilic properties of the deep well cell culture plate surface.
Test example 3:
impact Property measurement
The test method is carried out according to the specification of GB/T1843 plastics cantilever impact test method.
The deep well cell culture plates prepared in examples 1 to 8 and comparative example 1 were subjected to the above test, and the results are shown in Table 2:
TABLE 2 impact test results
| Sample of | Impact Strength (KJ/m) 2 ） |
| Comparative example 1 | 7.74 |
| Example 1 | 9.46 |
| Example 2 | 9.48 |
| Example 3 | 9.35 |
| Example 4 | 9.50 |
| Example 5 | 8.13 |
| Example 6 | 10.27 |
| Example 7 | 9.89 |
| Example 8 | 8.92 |
From the data analysis in Table 2, the impact strength of the deep hole cell plate prepared in example 1 is obviously higher than that of the deep hole cell plate prepared in example 5 and comparative example 1, and the effect of example 5 is better than that of comparative example 1, which shows that the chemical grafting modification of the deep hole cell plate by adopting itaconic acid derivatives can effectively enhance the impact strength of the deep hole cell culture plate and improve the toughness of the deep hole cell culture plate; meanwhile, under the condition that hexadiene diamine tartrate exists together, the toughness of the deep hole cell culture plate is better enhanced. The impact strength of the deep well cell plates prepared in example 6 was significantly higher than that of example 7, the effect of example 7 was slightly better than that of example 1, and the effect of example 8 was better than that of example 5, indicating that the addition of 3-bis (2-hydroxyethyl) amino-2-hydroxypropanesulfonic acid during the grafting treatment further increased the impact strength of the deep well cell culture plates.
Test example 4:
the deep-well cell culture plates prepared in examples 1 to 8 and comparative example 1 were tested for grafting efficiency, and the results are shown in Table 3:
TABLE 3 grafting test results
| Sample of | Grafting percentage (%) |
| Comparative example 1 | 0.45 |
| Example 1 | 0.47 |
| Example 2 | 0.46 |
| Example 3 | 0.45 |
| Example 4 | 0.46 |
| Example 5 | 0.45 |
| Example 6 | 0.73 |
| Example 7 | 0.54 |
| Example 8 | 0.50 |
From the data analysis in Table 3, it is evident that the grafting ratio of the deep-hole cell plate prepared in example 1 is equivalent to that of example 5 and comparative example 1, and the effect of example 5 is equivalent to that of comparative example 1, indicating that chemical grafting modification of the deep-hole cell plate with itaconic acid derivative does not negatively affect the grafting ratio of the deep-hole cell culture plate. The grafting rate of the deep hole cell plate prepared in example 6 is obviously higher than that of example 7, the effect of example 7 is slightly better than that of example 1, and the effect of example 8 is better than that of example 5, which shows that the grafting rate of the compound grafted on the deep hole cell culture plate can be effectively improved by adding 3-bis (2-hydroxyethyl) amino-2-hydroxy propane sulfonic acid in the grafting treatment process.
The conventional technology in the above embodiments is known to those skilled in the art, and thus is not described in detail herein.
The foregoing is merely illustrative of the present invention, and the present invention is not limited thereto, and any person skilled in the art will readily recognize that variations or substitutions are within the scope of the present invention. Therefore, the protection scope of the present invention shall be subject to the protection scope of the claims.
Claims (5)
1. A deep-well cell culture plate, comprising a basic deep-well cell plate and a surface-modified compound; the compound is connected to the surface of the deep hole cell plate through a chemical bond; the compound comprises hexadiene diamine tartrate and itaconic acid derivatives; the chemical structure of the itaconic acid derivative is shown as formula I:
the grafting rate of the compound on the surface of the deep hole cell plate is more than 0.4%;
the preparation method of the deep hole cell culture plate comprises the following steps:
pretreating, namely placing the deep-hole cell plate in an ultrasonic condition, and cleaning the deep-hole cell plate by sequentially adopting distilled water and acetone to obtain a pretreated deep-hole cell plate;
plasma activation with Ar/O 2 Mixing low-temperature plasmas to perform activation treatment on the surface of the pretreated deep-hole cell plate to obtain an activated deep-hole cell plate;
grafting, namely soaking the activated deep-hole cell plate in an aqueous solution containing hexadiene diamine tartrate and itaconic acid derivatives for treatment, and grafting to obtain a deep-hole cell culture plate;
the Ar/O ratio is 2 Mixing low temperature plasma process parameters, comprising: ar/O 2 The flow ratio is 38-40/10-13 sccm, the treatment time is 4-6 min, the power is 280-320W, and the working air pressure is 75-85 Pa;
the concentration of hexadiene diamine tartrate in the aqueous solution is 40-45wt% and the concentration of itaconic acid derivatives is 20-25wt%.
2. The deep-well cell culture plate of claim 1, wherein the compound is modified to the surface of the deep-well cell plate by solid phase infiltration grafting.
3. The deep-well cell culture plate of claim 1, wherein the aqueous solution is replaced by a water/acetone mixed solution.
4. A deep well cell culture plate according to claim 3, wherein the water/acetone mixed solution is further added with 3-bis (2-hydroxyethyl) amino-2-hydroxypropanesulfonic acid.
5. The deep-well cell culture plate of claim 4, wherein the 3-bis (2-hydroxyethyl) amino-2-hydroxypropanesulfonic acid is added in an amount of 0.01 to 0.02mol/L; the volume ratio of the acetone to the water is 0.8-1.4:1.
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