CN110124543A - A kind of method and apparatus generating body phase nano grade air bubbles - Google Patents
A kind of method and apparatus generating body phase nano grade air bubbles Download PDFInfo
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Abstract
本发明涉及一种生成体相纳米级气泡的方法,液体的反应介质容置于一腔室中,向腔室中通入成分可控的气体进行加压,使得腔室内的压强大于1ATM且不超过20ATM,保持5‑60min,然后放气减压5s‑30min至常压,反应介质中气体溶解度降低逐渐形成气核,随着放气逐渐在反应介质的内部生成体相纳米级气泡。本发明还涉及一种生成体相纳米级气泡的装置,腔室通过增压阀连接不同的气体并通过放气阀与外界连通,通过该增压阀和放气阀来控制腔室中的压强,液体的反应介质容置于该腔室中并响应于压强的变化生成体相纳米级气泡,观测装置对体相纳米气泡进行观测。根据本发明的方法,可以产生成分确定的体相纳米气泡,具有很好的重现性,操作简便。
The invention relates to a method for generating bulk nano-scale bubbles. A liquid reaction medium is accommodated in a chamber, and a gas with a controllable composition is introduced into the chamber for pressurization, so that the pressure in the chamber is greater than 1ATM and does not exceed 1ATM. If it exceeds 20ATM, keep it for 5-60min, then deflate and depressurize for 5s-30min to normal pressure, the solubility of the gas in the reaction medium decreases and gradually forms gas nuclei, and gradually generates bulk nano-sized bubbles inside the reaction medium with degassing. The present invention also relates to a device for generating bulk nano-scale bubbles. The chamber is connected to different gases through a booster valve and communicated with the outside world through a gas release valve. The pressure in the chamber is controlled through the booster valve and the gas release valve. A liquid reaction medium is accommodated in the chamber and generates bulk nano-bubbles in response to changes in pressure, and the observation device observes the bulk nano-bubbles. According to the method of the invention, bulk nano-bubbles with definite components can be produced, have good reproducibility, and are easy to operate.
Description
技术领域technical field
本发明涉及物理化学中的纳米气泡,更具体地涉及一种生成体相纳米级气泡的方法和装置。The invention relates to nano-bubbles in physical chemistry, more particularly to a method and device for generating bulk nano-bubbles.
背景技术Background technique
最早的体相纳米气泡的报导研究来源于人们对超声和空化效应的研究。到2000年开始有了大量关于体相气泡的研究,这些研究关注体相气泡的物理性质和它在溶液中的浓度。目前,由于体相纳米气泡的存在可能对多个领域带来影响,越来越多的各种领域的科学研究人员开始对体相纳米气泡产生浓厚的兴趣。例如,体相纳米气泡应用于表面清洗领域,通过吸附被机械搅拌激起的污染,可以阻止它们再沉淀而达到清洗的效果。在水处理领域,由于纳米气泡直径更小,比表面更大,在水中存在的时间更长,从而可以有效的提高曝气效率,很大程度上提高了污水处理的效率并促进生态的恢复。在生物医学领域,纳米气泡的应用研究虽然才刚刚起步,但前景广阔,微纳米气泡在超声成像和超声造影剂具有较强的回波反射性能,能够显著增强医学超声检测信号,弥补了传统超声诊断技术的敏感性和分辨率不够高的不足,而微米气泡受限于其较大的尺寸,应用范围受限于其血管内,然而纳米气泡不仅可以穿过血管壁到达组织内部,还可以突破脑血屏障,可以作为新型的超声造影剂应用于分子成像以及用于药物运输。在癌症的诊断与靶向治疗上也具有明显的优势,纳米级的气泡可以到达组织内部,在人体温度范围内可以长时间稳定存在,纳米气泡穿过由于癌症病变而变大的毛细血管在癌症细胞处聚集,超声成像此时可以比较清楚的观察到,然后加大超声频率,气泡破裂,从而达到靶向治疗的作用。The earliest reported research on bulk nanobubbles comes from the research on ultrasonic and cavitation effects. Since 2000, there have been a large number of studies on bulk bubbles, which focus on the physical properties of bulk bubbles and their concentrations in solutions. At present, because the existence of bulk nanobubbles may have an impact on many fields, more and more scientific researchers in various fields have begun to have a strong interest in bulk nanobubbles. For example, bulk nanobubbles are used in the field of surface cleaning. By absorbing the pollution aroused by mechanical agitation, they can be prevented from re-precipitating to achieve the cleaning effect. In the field of water treatment, due to the smaller diameter and larger specific surface of nanobubbles, they exist in water for a longer time, which can effectively improve the aeration efficiency, greatly improve the efficiency of sewage treatment and promote ecological restoration. In the field of biomedicine, although the application research of nano-bubbles has just started, it has broad prospects. Micro-nano bubbles have strong echo reflection performance in ultrasonic imaging and ultrasonic contrast agents, which can significantly enhance medical ultrasonic detection signals and make up for traditional ultrasonic imaging. The sensitivity and resolution of diagnostic techniques are not high enough, and microbubbles are limited by their large size, and their application range is limited in their blood vessels. However, nanobubbles can not only pass through the blood vessel wall to reach the interior of the tissue, but also break through Brain-blood barrier can be used as a new type of ultrasound contrast agent for molecular imaging and drug delivery. It also has obvious advantages in the diagnosis and targeted therapy of cancer. Nano-scale bubbles can reach the interior of the tissue and can exist stably for a long time within the temperature range of the human body. Nano-bubbles pass through capillaries that become enlarged due to cancer lesions. Cells gather at this time, and ultrasound imaging can be clearly observed at this time, and then increase the frequency of ultrasound, and the bubbles will burst, so as to achieve the effect of targeted therapy.
然而,在理论上还很难解释纳米气泡的稳定存在。根据经典热力学的解释:气泡的体积越小,内部的压力就越大,而压力大必然导致气泡破裂。例如直径为10nm的气泡,根据Laplace方程计算出的它的内部压力为144大气压(1.44×107Pa)(用物理参数表示),这么高的内部压力使得气泡瞬间就会消失。另外,根据分子动力学模拟研究,室温下水中纳米气泡的存在时间仅为几个到几百个皮秒,在实验中往往是观察不到这么短时间内存在的纳米气泡。但近年有几个研究小组用原子力显微镜直接观察到了固液界面上的纳米气泡,这为纳米气泡的存在提供了直接证据。另外,利用冷冻刻蚀方法和中子衍射方法也预测出了纳米气泡的存在。所以,要系统、深入地研究纳米气泡,解决理论与实验的不一致,必须先建立一种简便的、可控的方法来形成和深入研究纳米气泡的稳定机制。另外,形成气泡的气体来源也很难控制,导致实验的重复性差。同时,纳米气泡有多种潜在的生物医学应用价值,为了应用于生物实验或者临床医学研究中,我们需要开发在实验室条件下可以产生清洁气体种类和比例可精确调控的纳米气泡水的方法。已有的方法大多是产生固液界面的纳米气泡。However, it is difficult to explain the stable existence of nanobubbles theoretically. According to the interpretation of classical thermodynamics: the smaller the volume of the bubble, the greater the internal pressure, and the high pressure will inevitably cause the bubble to burst. For example, for a bubble with a diameter of 10 nm, its internal pressure calculated according to the Laplace equation is 144 atmospheres (1.44×10 7 Pa) (expressed by physical parameters), such a high internal pressure makes the bubble disappear instantly. In addition, according to molecular dynamics simulation studies, the existence time of nanobubbles in water at room temperature is only several to hundreds of picoseconds, and nanobubbles existing in such a short period of time are often not observed in experiments. But in recent years, several research groups have directly observed nanobubbles on the solid-liquid interface with atomic force microscopy, which provides direct evidence for the existence of nanobubbles. In addition, the existence of nanobubbles was also predicted by cryo-etching method and neutron diffraction method. Therefore, in order to study nanobubbles systematically and deeply, and to resolve the inconsistency between theory and experiment, it is necessary to establish a simple and controllable method to form and deeply study the stability mechanism of nanobubbles. In addition, the source of the gas that forms the bubbles is also difficult to control, resulting in poor repeatability of the experiment. At the same time, nanobubbles have a variety of potential biomedical applications. In order to be applied to biological experiments or clinical medical research, we need to develop methods that can produce nanobubble water with clean gas types and ratios that can be precisely adjusted under laboratory conditions. Most of the existing methods are to generate nanobubbles at the solid-liquid interface.
发明内容Contents of the invention
为了解决上述现有技术存在的无法稳定性产生体相纳米气泡的问题,更好地促进纳米气泡在各个领域的重要应用,本发明旨在提供一种生成体相纳米级气泡的方法和装置。In order to solve the problem of inability to stably generate bulk nanobubbles in the prior art and better promote the important application of nanobubbles in various fields, the present invention aims to provide a method and device for generating bulk nanobubbles.
本发明所述的生成体相纳米级气泡的方法,其中,液体的反应介质容置于一腔室中,向腔室中通入成分可控的气体进行加压,使得腔室内的压强大于1ATM且不超过20ATM,保持5-60min,反应介质中气体溶解度增大,形成过饱和状态,然后放气减压5s-30min至常压,反应介质中气体溶解度降低逐渐形成气核,随着放气逐渐在反应介质的内部生成体相纳米级气泡。The method for generating bulk nano-scale bubbles according to the present invention, wherein the liquid reaction medium is accommodated in a chamber, and a gas with a controllable composition is introduced into the chamber for pressurization, so that the pressure in the chamber is greater than 1ATM And do not exceed 20ATM, keep for 5-60min, the solubility of gas in the reaction medium increases, forming a supersaturated state, then degassing and reducing pressure for 5s-30min to normal pressure, the solubility of gas in the reaction medium decreases and gradually forms gas nuclei, with the degassing Bulk nanoscale bubbles are gradually generated inside the reaction medium.
优选地,放气减压15min至常压。Preferably, deflate and decompress for 15 minutes to normal pressure.
优选地,该气体是单一气体或混合气体。优选地,该气体为Kr、H2、N2、He、Ne、Ar、Xe、CH4、O2、CH4、SF6、CF4、C5F12、和/或上述任何气体通过任意比例混合的混合气体。Preferably, the gas is a single gas or a mixture of gases. Preferably, the gas is Kr, H 2 , N 2 , He, Ne, Ar, Xe, CH 4 , O 2 , CH 4 , SF 6 , CF 4 , C 5 F 12 , and/or any of the above gases. Proportionally mixed gas mixture.
优选地,该反应介质为水、酸溶液、碱溶液、盐溶液、缓冲液、和/或蛋白溶液。优选地,该酸溶液的PH值为4-7的盐酸。优选地,该碱溶液为碱金属或碱土金属的碱溶液。优选地,该碱溶液为PH为7-10的氢氧化钠、氢化化钾、氢氧化钙、和/或氢氧化镁。优选地,该缓冲液为生物实验中常用的缓冲液。优选地,该缓冲液为PBS、TE、TAE、TBE、TBS、和/或Tris-Glycine。优选地,该蛋白溶液为血清白蛋白溶液。优选地,该蛋白溶液为牛血清白蛋白、和/或胃蛋白酶及溶菌酶。Preferably, the reaction medium is water, acid solution, alkali solution, salt solution, buffer, and/or protein solution. Preferably, the pH value of the acid solution is hydrochloric acid of 4-7. Preferably, the alkaline solution is an alkaline solution of alkali metal or alkaline earth metal. Preferably, the alkaline solution is sodium hydroxide, potassium hydride, calcium hydroxide, and/or magnesium hydroxide with a pH of 7-10. Preferably, the buffer is a buffer commonly used in biological experiments. Preferably, the buffer is PBS, TE, TAE, TBE, TBS, and/or Tris-Glycine. Preferably, the protein solution is a serum albumin solution. Preferably, the protein solution is bovine serum albumin, and/or pepsin and lysozyme.
优选地,反应介质预先经过脱气处理以除去其中溶解的气体,以得到脱气反应介质。该脱气反应介质可以作为加压后减压的对照组的参比溶液。Preferably, the reaction medium is pre-degassed to remove dissolved gas therein, so as to obtain a degassed reaction medium. The degassed reaction medium can be used as a reference solution for a control group that is pressurized and then depressurized.
优选地,在进行清洁操作时不仅保证干净无尘,还保证无微生物污染,可以获得医用级别清洁度的纳米气泡。特别地,通入水蒸气,完全杀死管路中霉菌的孢子,使得最难杀死的微生物得到清除,保证其它微生物完全被消灭,从而得到医用级别清洁度的体相纳米气泡。产生的气泡水符合国标规定的清洁度就可以保证生产所用的器械可以达到所需的清洁度(参见GB/T6682)。Preferably, the cleaning operation is not only clean and dust-free, but also free from microbial contamination, and nanobubbles with medical grade cleanliness can be obtained. In particular, water vapor is introduced to completely kill the mold spores in the pipeline, remove the most difficult to kill microorganisms, and ensure that other microorganisms are completely eliminated, so as to obtain bulk nanobubbles with medical-grade cleanliness. The generated sparkling water meets the cleanliness specified by the national standard to ensure that the equipment used in production can achieve the required cleanliness (see GB/T6682).
本发明所述的生成体相纳米级气泡的装置,包括液体的反应介质、腔室和观测装置,该腔室通过增压阀连接不同的气体并通过放气阀与外界连通,通过该增压阀和放气阀来控制腔室中的压强,液体的反应介质容置于该腔室中并响应于压强的变化生成体相纳米级气泡,观测装置对体相纳米气泡进行观测。The device for generating bulk nano-bubbles according to the present invention includes a liquid reaction medium, a chamber and an observation device. A valve and an air release valve are used to control the pressure in the chamber. The liquid reaction medium is accommodated in the chamber and generates bulk nano-bubbles in response to changes in pressure. The observation device observes the bulk nano-bubbles.
优选地,该观测装置为纳米示踪分析系统(Nanoparticle Tracking Analysis,NTA)。具体地,将加压减压完毕的水注入纳米示踪分析系统的样品池中,使用显微镜观察并使用高分辨率摄像机进行颗粒运动轨迹的记录,利用Stokes-Einstein方程即可得到样品中每个颗粒物的粒径,并可测得颗粒的真实浓度。Preferably, the observation device is a nanoparticle tracking analysis system (Nanoparticle Tracking Analysis, NTA). Specifically, the pressurized and decompressed water is injected into the sample pool of the nano-tracing analysis system, and a microscope is used to observe and a high-resolution camera is used to record the particle trajectory, and the Stokes-Einstein equation can be used to obtain the The particle size of the particle, and the real concentration of the particle can be measured.
优选地,观测装置根据体相纳米气泡的数量和粒径分布来调整腔室中的压强、保持时间和放气时间,以最终控制体相纳米气泡的数量和大小。例如,观测装置可以调整加压时间、和/或放气时间来控制体相纳米气泡的数量和大小。具体地,对照组中通入的气体为单一成份的高纯度气体及多种单一成份高纯度气体以任意比例混合的气体,与未通入气体的空白组作对照来反映生成的体相纳米气泡的数量和浓度分布。Preferably, the observation device adjusts the pressure, holding time and deflation time in the chamber according to the quantity and particle size distribution of bulk nanobubbles, so as to finally control the quantity and size of bulk nanobubbles. For example, the observation device can adjust the pressurization time, and/or deflation time to control the number and size of bulk nanobubbles. Specifically, the gas injected into the control group is a single-component high-purity gas or a gas mixed with a variety of single-component high-purity gases in any proportion. It is compared with the blank group without gas to reflect the generated bulk nanobubbles. quantity and concentration distribution.
根据本发明的方法,可以产生成分确定的、单一成分的体相纳米气泡或多种成分混合气体的纳米气泡,具有很好的重现性,操作简便。另外,根据本发明的方法产生的体相纳米气泡,清洁度高,可用于生物医学研究,甚至达到医用级别清洁度。而且,根据本发明的方法,可以控制纳米气泡的气体类型、纳米气泡的大小和数量。根据本发明的装置,可以进一步开发成医疗器械等各种新型设备,在诸多领域都有广泛的应用。According to the method of the invention, single-component bulk nano-bubbles or multi-component mixed gas nano-bubbles can be produced, which has good reproducibility and is easy to operate. In addition, the bulk nano-bubbles produced by the method of the present invention have high cleanliness, can be used in biomedical research, and even reach medical grade cleanliness. Moreover, according to the method of the present invention, the gas type of the nanobubbles, the size and the number of the nanobubbles can be controlled. According to the device of the present invention, it can be further developed into various new equipment such as medical equipment, and has wide application in many fields.
附图说明Description of drawings
图1是根据本发明的实施例1的体相纳米气泡的数量分布图;Fig. 1 is the quantity distribution figure of the bulk phase nanobubble according to embodiment 1 of the present invention;
图2A是根据本发明的实施例2的通气体加压前的缓冲溶液的镜下结果图;Fig. 2A is a microscopic result diagram of the buffer solution before pressurization of the ventilator according to Example 2 of the present invention;
图2B是根据本发明的实施例2的通气体加压后的缓冲溶液的镜下结果图;Fig. 2B is a microscopic result diagram of the buffer solution pressurized by the ventilator according to Example 2 of the present invention;
图2C是根据本发明的实施例2的通气体加压产生纳米气泡之后再脱气的缓冲溶液的镜下结果图;Fig. 2C is a microscopic result diagram of a buffer solution degassed after nanobubbles are generated by pressurizing the ventilator according to Example 2 of the present invention;
图3示出了根据本发明的实施例2的通气体加压前、加压后、加压产生纳米气泡之后再脱气的缓冲溶液中的体相纳米气泡的粒径分布与纳米气泡浓度;Fig. 3 shows the particle size distribution and nanobubble concentration of the bulk nanobubbles in the buffer solution degassed after the venting gas is pressurized, pressurized, and pressurized to generate nanobubbles according to Embodiment 2 of the present invention;
图4示出了根据本发明的实施例2的通气体加压前、加压后、加压产生纳米气泡之后再脱气的缓冲溶液中的体相纳米气泡的数量分布;Fig. 4 shows the number distribution of the bulk nanobubbles in the buffer solution degassed after the pressurization of the ventilating gas according to Embodiment 2 of the present invention before, after pressurization, and after nanobubbles are generated;
图5示出了根据本发明的实施例5的牛血清白蛋白溶液中的体相纳米气泡。FIG. 5 shows bulk nanobubbles in bovine serum albumin solution according to Example 5 of the present invention.
具体实施方式Detailed ways
下面结合附图,给出本发明的较佳实施例,并予以详细描述。Below in conjunction with the drawings, preferred embodiments of the present invention are given and described in detail.
实施例中所用纳米颗粒分析仪是NanoSight NS300系统(Malvern Instruments,Inc.)、配备样品池(蓝色激光器)、显微镜机架、高分辨率摄像机sCMOS、纳米示踪分析系统;水是Millipore超纯水,样品池在使用前用水、乙醇、水交替清洗。带盖玻璃试剂瓶及玻璃注射器在使用前用去离子水清洗3遍并使用超声洗净,在干燥箱中烘干。The nanoparticle analyzer used in the embodiment is NanoSight NS300 system (Malvern Instruments, Inc.), equipped with sample cell (blue laser), microscope frame, high-resolution camera sCMOS, nano tracer analysis system; water is Millipore ultrapure Water, the sample cell is washed alternately with water, ethanol, and water before use. Before use, the glass reagent bottle and the glass syringe were washed with deionized water three times, cleaned with ultrasonic waves, and dried in a drying oven.
制作脱气水,使用脱气水来配置各种液体的反应介质。制作脱气水的步骤:取水置于洁净的带盖玻璃试剂瓶中,冰冻。冰冻的目的是尽可能除去溶解到水中的气体,进行深度脱气。取下盖子,使用封口膜封口,并在封口膜上打四个小孔,置于真空干燥箱中,抽真空,0.1ATM保持10个小时,取出重复上述操作三遍,溶液选择为脱气水,配置其它溶液要注意为非腐蚀性,腐蚀性溶液会腐蚀载物台上的镀层,使其不平整,不平整的镀层会折射激光,使信噪比降低。Make degassed water, use degassed water to configure various liquid reaction media. Steps for making degassed water: Take water, put it in a clean glass reagent bottle with a lid, and freeze it. The purpose of freezing is to remove the gas dissolved in the water as much as possible and carry out deep degassing. Take off the cover, seal it with a parafilm, and punch four small holes on the parafilm, place it in a vacuum drying oven, evacuate it, keep it at 0.1ATM for 10 hours, take it out and repeat the above operation three times, the solution is degassed water , the configuration of other solutions should be non-corrosive. Corrosive solutions will corrode the coating on the stage and make it uneven. The uneven coating will refract the laser and reduce the signal-to-noise ratio.
实施例1在脱气水中生成氪气纳米气泡Embodiment 1 generates krypton nano-bubbles in degassed water
在脱气水中加氪气到6个标准大气压保持30min,得到的体相纳米气泡的数量分布如图1所示,气泡数量每ml可以达108量级。Add krypton gas to the deaerated water to 6 standard atmospheres and keep it for 30 minutes. The distribution of the number of nanobubbles in the bulk phase is shown in Figure 1. The number of bubbles per ml can reach the order of 10 8 .
纳米气泡数量具体测量步骤如下:The specific measurement steps for the number of nanobubbles are as follows:
打开Nanosight启动温控软件NTA temperature controller;检查是否连接完好,开启主程序NTA program。用洁净的玻璃注射器取用加压减压法制备的纳米气泡水,吸入液体量约1ml左右,排出注射器顶端的空气,“缓慢”注入样品池中,至“出口端”有液体流出为止。为防止出口处液体滴下,垫小块卷纸。将样品池置于显微镜载物台上,蓝色激光应为发光状态(若未亮点击NTA软件Capture界面中camera level调至较高处,激发激光),前后、左右调节样品池位置,上下调节焦距,至能观测到蓝色激光束,利用NTA软件的观测图像精细调节焦距、位置。若视野一片高亮,说明样品太浓,需要重新稀释。高亮需要稀释,设置Camera duration为60s,记录5次取均值。如图1所示,在时间范围5s<t≦15min内,随着放气时间越长,生成的纳米气泡数量越多。将纳米气泡水置于真空干燥箱中脱气之后测量发现纳米气泡的数量显著下降,所以可以说明实验中所生成的气泡成分为气体,至于气体成分,因为气体来源是我们直接通入的可控的特定气体成分。且加压是通过增加体积固定的腔体中的气体含量来实现的,在加压之前先使用反应气体对腔体内进行吹扫,吹出样品腔中的空气。说明生成的气泡为特定成分可控的气体纳米气泡,而不是其它物质。Open Nanosight and start the temperature control software NTA temperature controller; check whether the connection is intact, and start the main program NTA program. Use a clean glass syringe to take the nano-bubble water prepared by the pressurization and decompression method, inhale about 1ml of the liquid, discharge the air at the top of the syringe, and "slowly" inject it into the sample pool until the liquid flows out from the "outlet end". To prevent the liquid from dripping at the outlet, place a small piece of roll paper. Place the sample cell on the microscope stage, the blue laser light should be in the luminous state (if it is not on, click the camera level in the Capture interface of the NTA software to adjust to a higher level to activate the laser), adjust the position of the sample cell front and back, left and right, up and down The focal length is until the blue laser beam can be observed, and the focal length and position can be finely adjusted by using the observation image of the NTA software. If the field of view is bright, the sample is too concentrated and needs to be re-diluted. The highlight needs to be diluted, set the Camera duration to 60s, and record 5 times to get the average value. As shown in Figure 1, within the time range of 5s<t≦15min, the longer the deflation time, the greater the number of nanobubbles generated. After degassing the nano-bubble water in a vacuum drying oven, it was found that the number of nano-bubbles decreased significantly, so it can be explained that the bubbles generated in the experiment are gas. specific gas components. In addition, the pressurization is realized by increasing the gas content in the cavity with a fixed volume. Before pressurization, the cavity is purged with a reaction gas to blow out the air in the sample cavity. It shows that the generated bubbles are gas nano-bubbles with controllable specific composition, rather than other substances.
观察过程如上,设置camera level为9,屏幕亮度为9,颗粒粒径阈值为5。随后取剩余的纳米气泡水,使用封口膜封口,扎若干个小洞,置于真空干燥箱中脱气一小时。观察过程如上,设置camera level为8,屏幕亮度为10,颗粒粒径阈值为5。The observation process is as above, set the camera level to 9, the screen brightness to 9, and the particle size threshold to 5. Then take the remaining nano-bubble water, seal it with a parafilm, make several small holes, and place it in a vacuum drying oven for degassing for one hour. The observation process is as above, set the camera level to 8, the screen brightness to 10, and the particle size threshold to 5.
实施例2在TBE溶液中生成氪气纳米气泡Embodiment 2 generates krypton nano bubbles in TBE solution
用去离子水配置TBE溶液,使用玻璃注射器取玻璃试剂瓶中距离气液界面2cm左右溶液,注入纳米示踪分子系统的样品池中。设置camera level为8,屏幕亮度为10,颗粒粒径阈值为5。随后制作纳米气泡水,使用移液器取3mL TBE溶液置于三通阀的样品池中,通入氪气使腔体中的压强达到6个标准大气压,保持30分钟,之后缓慢放气过程持续15分钟。Prepare the TBE solution with deionized water, use a glass syringe to take the solution in the glass reagent bottle about 2cm away from the gas-liquid interface, and inject it into the sample cell of the nanotracer molecular system. Set the camera level to 8, the screen brightness to 10, and the particle size threshold to 5. Then make nano-bubble water, use a pipette to take 3mL of TBE solution and place it in the sample pool of the three-way valve, introduce krypton gas to make the pressure in the cavity reach 6 standard atmospheres, keep it for 30 minutes, and then the slow degassing process continues 15 minutes.
观察发现,加压减压之后的TBE溶液中颗粒数显著增加,达1.6×108个/ml,脱气之后颗粒数显著减少,减少至1.2×107个/ml,甚至少于初始未经过任何处理的TBE溶液中的颗粒数,即1.3×107/ml。说明通过加压减压法产生的氮气纳米气泡TBE溶液中增加的颗粒为纳米气泡,脱气后颗粒数显著减少说明纳米气泡TBE溶液中增加的纳米气泡中的成分确实为气体。It was observed that the number of particles in the TBE solution after pressurization and decompression increased significantly to 1.6×10 8 /ml, and the number of particles decreased significantly after degassing to 1.2×10 7 /ml, which was even less than that of the initial untreated solution. The number of particles in any treated TBE solution, ie 1.3 x 10 7 /ml. It shows that the particles increased in the nitrogen nanobubble TBE solution produced by the pressurization and decompression method are nanobubbles, and the number of particles decreases significantly after degassing, which shows that the composition of the nanobubbles increased in the nanobubble TBE solution is indeed gas.
如图2A-2C分别通气体加压前的TBE缓冲溶液、通气体加压后的TBE缓冲溶液和通气体加压产生纳米气泡之后再脱气的TBE缓冲溶液。通过高速摄影机记录纳米气泡的运动轨迹,取其中一帧,NTA获取纳米气泡的尺寸分布以及浓度是通过高速摄影机记录纳米气泡折射光斑运动轨迹,根据运动速度利用爱因斯坦-斯托克斯方程求解而得的。从图中可以看出未经过加压减压处理的TBE缓冲溶液中纳米气泡数量不多,而经过加压减压处理的TBE缓冲溶液中纳米气泡数量显著增加,图2C在图2B基础上进行减压深度脱气处理,纳米气泡数量又显著减少。图3示出了通气体加压前、加压后、加压产生纳米气泡之后再脱气的TBE缓冲溶液中的体相纳米气泡的粒径分布与纳米气泡浓度,如图所示,未经过处理的水中颗粒数小于105量级,而经过加压减压法处理生成的氪气纳米气泡水中纳米气泡的含量显著增加,粒径为~100nm的纳米气泡数量接近106,平均粒径在100nm-200nm之间的纳米气泡数量接近2.0×106个/ml,平均粒径接近200nm的纳米气泡数量远超过106。图4示出了通气体加压前、加压后、加压产生纳米气泡之后再脱气的TBE缓冲溶液中的体相纳米气泡的数量分布,加压前溶液中可以探测到的纳米颗粒很少,而经过加压减压法处理之后,纳米气泡的数量显著增多,数量超过的106,比未处理前增加了一个数量级,而为了证明增加的这些纳米颗粒都是纳米气泡,我们通过脱气,将气泡水中的气泡抽掉,溶液中的纳米颗粒的数量又显著降低,降到和使用加压减压法处理之前一个数量级,说明了使用此方法可以产生大量的纳米气泡,多出的一个数量级的纳米颗粒即为纳米气泡。As shown in Figures 2A-2C respectively, the TBE buffer solution before gas pressurization, the TBE buffer solution after gas pressurization, and the TBE buffer solution degassed after gas pressurization to generate nanobubbles. The trajectory of the nanobubbles is recorded by a high-speed camera, and one frame is taken, and the size distribution and concentration of the nanobubbles are obtained by NTA. The trajectory of the refracted spot of the nanobubbles is recorded by a high-speed camera, and the Einstein-Stokes equation is used to solve the problem according to the speed of movement. And got it. It can be seen from the figure that the number of nanobubbles in the TBE buffer solution that has not been subjected to pressure and decompression treatment is small, but the number of nanobubbles in the TBE buffer solution that has been subjected to pressure and decompression treatment increases significantly. Figure 2C is based on Figure 2B After decompression and deep degassing treatment, the number of nano bubbles is significantly reduced. Figure 3 shows the particle size distribution and nanobubble concentration of the bulk nanobubbles in the degassed TBE buffer solution before, after pressurization, after pressurization to generate nanobubbles, as shown in the figure, without The number of particles in the treated water is less than 10 5 , while the content of nano-bubbles in krypton nano-bubble water generated by the pressure and decompression method is significantly increased. The number of nanobubbles between 100nm and 200nm is close to 2.0×10 6 /ml, and the number of nanobubbles with an average particle size close to 200nm is far more than 10 6 . Figure 4 shows the distribution of the number of bulk nanobubbles in the degassed TBE buffer solution before, after pressurization, and after pressurization to generate nanobubbles. The nanoparticles that can be detected in the solution before pressurization are very small. However, after the pressure and decompression treatment, the number of nanobubbles increased significantly, and the number exceeded 10 6 , which was an order of magnitude higher than that before untreated. In order to prove that the increased nanoparticles are all nanobubbles, we used the method of decompression Gas, pump out the bubbles in the bubble water, the number of nanoparticles in the solution is significantly reduced, down to an order of magnitude before using the pressure and decompression method, which shows that this method can produce a large number of nanobubbles, and the extra Nanoparticles of an order of magnitude are nanobubbles.
实施例3在稀氢氧化钾溶液中生成氮气纳米气泡Embodiment 3 generates nitrogen nanobubbles in dilute potassium hydroxide solution
装置和操作步骤如实施例1,但使用0.001-1.0mol/L的氢氧化钾溶液作为溶剂,加6个标准大气压,保持30分钟,缓慢放气时间持续15秒,得到的纳米气泡粒径范围为50~300nm。气泡数量可达2.5×108个/ml,误差范围为±5.9×107个/ml。The device and operation steps are as in Example 1, but use 0.001-1.0mol/L potassium hydroxide solution as a solvent, add 6 standard atmospheric pressure, keep for 30 minutes, slow deflation time continues for 15 seconds, and the obtained nanobubble particle size range 50-300nm. The number of bubbles can reach 2.5×10 8 /ml, and the error range is ±5.9×10 7 /ml.
实施例4在氯化钠溶液中生成氮气纳米气泡Embodiment 4 generates nitrogen nanobubbles in sodium chloride solution
装置和操作步骤如实施例1,但使用0.001-0.1mol/L的氯化钠溶液,生成的纳米气泡大小与TBE和氢氧化钾溶液的类似。气泡数量可达2.13×108个/ml,误差范围为±6.6×107个/ml。The device and operation steps are as in Example 1, but using 0.001-0.1 mol/L sodium chloride solution, the size of the generated nanobubbles is similar to that of TBE and potassium hydroxide solution. The number of bubbles can reach 2.13×10 8 /ml, and the error range is ±6.6×10 7 /ml.
实施例5在BSA(牛血清白蛋白)溶液中生成氮气纳米气泡Embodiment 5 generates nitrogen nanobubbles in BSA (bovine serum albumin) solution
加压向0.1mg/mL的BSA溶液中通入氮气,使加压减压装置内部压强达到6ATM,保持30分钟,放气5分钟所产生的纳米气泡的粒径和数量分布,如图5所示,对牛血清白蛋白溶液通过加压减压的方法也可以产生大量的纳米气泡。没有经过加压减压法处理的牛血清白蛋白溶液中的颗粒数作为空白背景来对比,显示出牛血清白蛋白溶液中通过加压减压可以产生大量的纳米气泡。Pressurize and feed nitrogen into the BSA solution of 0.1mg/mL, so that the internal pressure of the pressurization and decompression device reaches 6ATM, keep it for 30 minutes, and release the gas for 5 minutes. The particle size and number distribution of the nanobubbles produced, as shown in Figure 5 Show, also can produce a large amount of nano-bubbles to bovine serum albumin solution by the method of pressurization and decompression. The number of particles in the bovine serum albumin solution that has not been processed by the pressurization and decompression method is compared as a blank background, showing that a large number of nano bubbles can be produced in the bovine serum albumin solution by pressurization and decompression.
以上所述的,仅为本发明的较佳实施例,并非用以限定本发明的范围,本发明的上述实施例还可以做出各种变化。即凡是依据本发明申请的权利要求书及说明书内容所作的简单、等效变化与修饰,皆落入本发明专利的权利要求保护范围。本发明未详尽描述的均为常规技术内容。What is described above is only a preferred embodiment of the present invention, and is not intended to limit the scope of the present invention. Various changes can also be made to the above embodiments of the present invention. That is to say, all simple and equivalent changes and modifications made according to the claims and description of the application for the present invention fall within the protection scope of the claims of the patent of the present invention. What is not described in detail in the present invention is conventional technical contents.
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