CN110438002A - The wireless multi-channel system of temperature monitoring during a kind of adherent growth for cell - Google Patents
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Abstract
本发明涉及一种用于细胞贴壁生长过程中温度监测的无线多通道系统，包括恒温培养箱、薄膜铂热电阻传感器、细胞培养板、3D打印框体以及电阻检测装置，所述细胞培养板放置在恒温培养箱内，所述细胞培养板上设有培养孔，所述培养孔内设有薄膜铂热电阻传感器以及3D打印框体，所述薄膜铂热电阻传感器通过3D打印框体与细胞培养板相连；所述薄膜铂热电阻传感器与电阻检测装置相连；本发明公开了一种用于细胞贴壁生长过程中温度监测的无线多通道系统，在薄膜铂热电阻传感器表面接种细胞，在细胞培养的同时，无线监控细胞温度变化情况，不受中间介质的干扰，生物相容性好，对细胞无创。
The invention relates to a wireless multi-channel system for temperature monitoring in the process of cell adherent growth, which includes a constant temperature incubator, a thin-film platinum thermal resistance sensor, a cell culture plate, a 3D printing frame and a resistance detection device. The cell culture plate Placed in a constant temperature incubator, the cell culture plate is provided with a culture hole, and a thin-film platinum thermal resistance sensor and a 3D printing frame are arranged in the culture hole, and the thin-film platinum thermal resistance sensor is connected to the cell through the 3D printing frame. The culture plate is connected; the thin-film platinum thermal resistance sensor is connected with the resistance detection device; the invention discloses a wireless multi-channel system for temperature monitoring in the process of cell adherent growth, in which cells are seeded on the surface of the thin-film platinum thermal resistance sensor, and the While culturing cells, wirelessly monitor cell temperature changes without interference from intermediate media, have good biocompatibility, and are non-invasive to cells.
Description
技术领域technical field
本发明涉及一种用于细胞贴壁生长过程中温度监测的无线多通道系统，属于细胞温度监测系统领域。The invention relates to a wireless multi-channel system for temperature monitoring in the process of cell adherent growth, belonging to the field of cell temperature monitoring systems.
背景技术Background technique
温度是细胞活动最重要和最基本的生理参数，细胞的每个生理过程都伴随着温度的变化，有研究表明，随着人类多能干细胞的分化，其主要代谢方式也逐渐从糖酵解向氧化磷酸化转变，这种转变也反应在细胞温度变化层面。同时有研究表明，外界温度对调控细胞活动也具有显著效果，对不同的细胞，其调节效果各不相同，例如热刺激可促进成骨细胞增殖分化，同时热刺激也会降低大鼠睾丸支持细胞的增殖活力，因此细胞生长过程中的温度检测，对生命科学的研究具有重要意义。Temperature is the most important and basic physiological parameter for cell activities. Every physiological process of cells is accompanied by changes in temperature. Studies have shown that with the differentiation of human pluripotent stem cells, their main metabolic mode gradually shifts from glycolysis to Oxidative phosphorylation shifts, which are also reflected in changes in cellular temperature. At the same time, studies have shown that external temperature also has a significant effect on the regulation of cell activity, and the regulatory effects are different for different cells. For example, heat stimulation can promote the proliferation and differentiation of osteoblasts, and heat stimulation can also reduce the growth rate of rat testicular Sertoli cells. Therefore, the temperature detection during cell growth is of great significance to the research of life science.
目前针对细胞温度进行测量的技术主要有：荧光纳米温度计、扫描热探针、微纳热电偶探针。At present, the technologies for measuring cell temperature mainly include: fluorescent nanothermometer, scanning thermal probe, and micro-nano thermocouple probe.
荧光纳米温度计，利用量子点荧光标记探针、温敏荧光纳米颗粒、绿色荧光蛋白等材料，对活细胞进行标记，通过荧光信号的波长、强度、寿命等特性的变化，反应活细胞温度的变化。荧光纳米温度计具有较高的空间分辨率和温度变化灵敏度，但很难做到实时监测，且在重复性和生物相容性方面还存在不足。Fluorescent nano-thermometers use quantum dot fluorescent labeling probes, temperature-sensitive fluorescent nanoparticles, green fluorescent protein and other materials to label living cells, and reflect changes in the temperature of living cells through changes in the wavelength, intensity, and lifetime of fluorescent signals. . Fluorescent nanothermometers have high spatial resolution and sensitivity to temperature changes, but it is difficult to monitor in real time, and there are still deficiencies in repeatability and biocompatibility.
扫描热探针，在STM（扫描隧道显微镜）扫描探针的顶端，发展一个基于热电效应的温度传感器，能够在纳米量级进行物体表面温度成像，其对设备条件以及操作人员技术要求较高，且不适合作为温度实时监控手段。The scanning thermal probe, on the top of the STM (scanning tunneling microscope) scanning probe, develops a temperature sensor based on the pyroelectric effect, which can image the surface temperature of the object at the nanometer level, which requires high equipment conditions and operator skills. And it is not suitable as a means for real-time temperature monitoring.
微纳热电偶探针，基于金属塞贝克效应，把温度信号转换成热电动势信号，适合于检测单细胞内温度变化，具有测量精度高、测量温度范围宽的优点，但使用时需要穿刺细胞，属于有创检测，只能测量单个细胞的温度变化，且测温时间较短。The micro-nano thermocouple probe, based on the metal Seebeck effect, converts the temperature signal into a thermoelectromotive force signal, which is suitable for detecting temperature changes in a single cell. It has the advantages of high measurement accuracy and a wide temperature range, but it needs to puncture the cell when using it. It is an invasive test that can only measure the temperature change of a single cell, and the temperature measurement time is short.
发明内容Contents of the invention
本发明针对现有技术中存在的不足，提供了一种用于细胞贴壁生长过程中温度监测的无线多通道系统，以解决现有技术中存在的问题。Aiming at the deficiencies in the prior art, the invention provides a wireless multi-channel system for temperature monitoring in the process of cell adherent growth, so as to solve the problems in the prior art.
为实现上述目的，本发明采用的技术方案如下：To achieve the above object, the technical scheme adopted in the present invention is as follows:
一种用于细胞贴壁生长过程中温度监测的无线多通道系统，包括恒温培养箱、薄膜铂热电阻传感器、细胞培养板、3D打印框体以及电阻检测装置，所述细胞培养板放置在恒温培养箱内，所述细胞培养板上设有培养孔，所述培养孔内设有薄膜铂热电阻传感器以及3D打印框体，所述薄膜铂热电阻传感器通过3D打印框体与细胞培养板相连；所述薄膜铂热电阻传感器与电阻检测装置相连。A wireless multi-channel system for temperature monitoring during cell adherent growth, including a constant temperature incubator, a thin-film platinum thermal resistance sensor, a cell culture plate, a 3D printing frame, and a resistance detection device. The cell culture plate is placed in a constant temperature In the incubator, the cell culture plate is provided with a culture hole, the culture hole is provided with a thin-film platinum thermal resistance sensor and a 3D printing frame, and the thin-film platinum thermal resistance sensor is connected to the cell culture plate through the 3D printing frame ; The thin-film platinum thermal resistance sensor is connected with the resistance detection device.
作为本发明的一种改进，所述细胞培养板上设有与薄膜铂热电阻传感器相对应的通孔；所述3D打印框体内设有与薄膜铂热电阻传感器相对应的凹槽。As an improvement of the present invention, the cell culture plate is provided with a through hole corresponding to the thin-film platinum thermal resistance sensor; the 3D printing frame is provided with a groove corresponding to the thin-film platinum thermal resistance sensor.
作为本发明的一种改进，所述薄膜铂热电阻传感器的数量为至少12个，所述培养孔的数量为至少12个。As an improvement of the present invention, the number of the thin-film platinum thermal resistance sensors is at least 12, and the number of the culture wells is at least 12.
作为本发明的一种改进，还包括计算机，所述计算机通过无线网络终端与电阻检测装置相连。计算机内加载有上位机监测系统，由校准曲线参数、温度监测界面、温度监测历史三个界面组成。上位机监测系统接收电阻检测装置数据，经过处理后，实时显示每组传感器采集到的温度信息，并进行数据记录和温度监测历史绘制。As an improvement of the present invention, a computer is also included, and the computer is connected to the resistance detection device through a wireless network terminal. The computer is loaded with a host computer monitoring system, which consists of three interfaces: calibration curve parameters, temperature monitoring interface, and temperature monitoring history. The host computer monitoring system receives the data of the resistance detection device, and after processing, displays the temperature information collected by each group of sensors in real time, and performs data recording and temperature monitoring history drawing.
作为本发明的一种改进，所述无线网络终端为蓝牙。As an improvement of the present invention, the wireless network terminal is bluetooth.
作为本发明的一种改进，还包括锂电池，所述锂电池设置在恒温培养箱内，所述锂电池与电阻检测装置相连。As an improvement of the present invention, a lithium battery is also included, the lithium battery is arranged in a constant temperature incubator, and the lithium battery is connected with a resistance detection device.
作为本发明的一种改进，所述3D打印框体为白色光敏树脂材料通过立体光固化成型工艺打印。As an improvement of the present invention, the 3D printing frame is printed with a white photosensitive resin material through a stereolithography molding process.
作为本发明的一种改进，所述的一种用于细胞贴壁生长过程中温度监测的无线多通道在测定的细胞中的应用。As an improvement of the present invention, the application of the wireless multi-channel for temperature monitoring in the process of cell adherent growth to the measured cells.
作为本发明的一种改进，所述细胞包括肿瘤细胞、干细胞、内皮细胞以及神经细胞。As an improvement of the present invention, the cells include tumor cells, stem cells, endothelial cells and nerve cells.
作为本发明的一种改进，所述细胞的测定方法，包括如下步骤：As an improvement of the present invention, the cell assay method includes the following steps:
步骤一：使用生物材料对薄膜铂热电阻传感器进行包被处理，在超净台中过夜晾干；Step 1: Use biological materials to coat the thin-film platinum thermal resistance sensor, and dry it overnight in an ultra-clean bench;
步骤二：使用硅酮导热胶将薄膜铂热电阻传感器、3D打印框体、细胞培养板固定，构建细胞培养环境；Step 2: Use silicone heat-conducting glue to fix the thin-film platinum thermal resistance sensor, 3D printing frame, and cell culture plate to build a cell culture environment;
步骤三：在常温下使用环氧乙烷气体对设备进行广谱灭菌；Step 3: Broad-spectrum sterilization of the equipment with ethylene oxide gas at room temperature;
步骤四：将浓度为5×108 /L的细胞溶液接种到3D打印框体中，在细胞培养板中加入培养液，放入恒温培养箱中培养；Step 4: Inoculate the cell solution with a concentration of 5×10 8 /L into the 3D printing frame, add the culture medium to the cell culture plate, and place it in a constant temperature incubator for cultivation;
步骤五：开启计算机，通过蓝牙传输数据，实时观察其温度变化情况。Step 5: Turn on the computer, transmit data via Bluetooth, and observe its temperature changes in real time.
薄膜铂热电阻传感器，利用铂热电阻感应细胞贴壁生长期间温度变化；3D打印框体，用于固定铂热电阻传感器，内部空间提供细胞培养环境；电阻检测装置，用于测量多组薄膜铂热电阻传感器的阻值及数据传输；计算机，用于数据接收、处理、保存，并实时显示其温度变化。所述薄膜铂热电阻传感器可诱导细胞贴附传感器表面生长。Thin-film platinum thermal resistance sensor, using platinum thermal resistance to sense temperature changes during cell adherent growth; 3D printing frame, used to fix platinum thermal resistance sensor, and the internal space provides a cell culture environment; resistance detection device, used to measure multiple groups of thin-film platinum thermal resistance sensors The resistance value and data transmission of the thermal resistance sensor; the computer is used for data reception, processing, storage, and real-time display of its temperature change. The thin-film platinum thermal resistance sensor can induce cell-attached sensor surface growth.
所述电阻检测系统具有多个检测通道，并采用开尔文四线制电阻检测电路。所述电阻检测系统使用7.4V锂电池供电，并通过蓝牙模块进行数据传输，实现无线便携。所述计算机通过蓝牙接收数据，对数据转化、分析、保存，并在上位机实时显示其温度变化情况。The resistance detection system has multiple detection channels and adopts a Kelvin four-wire resistance detection circuit. The resistance detection system is powered by a 7.4V lithium battery, and transmits data through a Bluetooth module to realize wireless portability. The computer receives data through bluetooth, converts, analyzes and saves the data, and displays its temperature change in real time on the upper computer.
由于采用了以上技术，本发明较现有技术相比，具有的有益效果如下：Owing to having adopted above technique, the present invention compares with prior art, has the beneficial effect as follows:
本发明公开了一种用于细胞贴壁生长过程中温度监测的无线多通道系统，在薄膜铂热电阻传感器表面接种细胞，在细胞培养的同时，无线监控细胞温度变化情况，不受中间介质的干扰，生物相容性好，对细胞无创；一块细胞培养板上可以同时设置多个实验组和多个对照组，可重复性好。测量设备使用锂电池供电，可实现24小时以上的无线温度监测，通过上位机数据处理系统可实时观察温度变化情况，方便实验人员观察。The invention discloses a wireless multi-channel system for temperature monitoring in the process of cell adherent growth. Cells are seeded on the surface of a thin-film platinum thermal resistance sensor, and the temperature change of the cells is wirelessly monitored while the cells are being cultivated without being affected by an intermediate medium. Interference, good biocompatibility, and non-invasive to cells; multiple experimental groups and multiple control groups can be set on one cell culture plate at the same time, with good repeatability. The measuring equipment is powered by a lithium battery, which can realize wireless temperature monitoring for more than 24 hours. The temperature change can be observed in real time through the host computer data processing system, which is convenient for experimenters to observe.
附图说明Description of drawings
图1是细胞培养板的结构示意图；Fig. 1 is the structural representation of cell culture plate;
图2是一种用于细胞贴壁生长过程中温度监测的无线多通道系统的示意图；Fig. 2 is a schematic diagram of a wireless multi-channel system used for temperature monitoring during cell adherent growth;
图3为24小时内神经干细胞温度和对照组温度（环境温度）变化；Figure 3 is the change of neural stem cell temperature and control group temperature (environmental temperature) within 24 hours;
图4为诱导神经干细胞贴壁过程中，实验组与对照组温度差别变化；Figure 4 is the temperature difference between the experimental group and the control group during the process of inducing neural stem cell attachment;
图中：1、计算机，2、恒温培养箱，3、锂电池，4、蓝牙，5、电阻检测装置，6、细胞培养板，7、3D打印框体，8、薄膜铂热电阻传感器，9、凹槽，10、通孔，11、培养孔。In the figure: 1. Computer, 2. Constant temperature incubator, 3. Lithium battery, 4. Bluetooth, 5. Resistance detection device, 6. Cell culture plate, 7. 3D printing frame, 8. Thin film platinum thermal resistance sensor, 9 , groove, 10, through hole, 11, culture hole.
具体实施方式Detailed ways
下面结合附图和具体实施方式，进一步阐明本发明。The present invention will be further explained below in conjunction with the accompanying drawings and specific embodiments.
结合附图可见，一种用于细胞贴壁生长过程中温度监测的无线多通道系统，包括恒温培养箱2、薄膜铂热电阻传感器8、细胞培养板6、3D打印框体7以及电阻检测装置5，所述细胞培养板6放置在恒温培养箱2内，所述细胞培养板6上设有培养孔11，所述培养孔11内设有薄膜铂热电阻传感器8以及3D打印框体7，所述薄膜铂热电阻传感器8通过3D打印框体7与细胞培养板6相连；所述薄膜铂热电阻传感器8与电阻检测装置5相连。It can be seen from the accompanying drawings that a wireless multi-channel system for temperature monitoring during cell adherent growth includes a constant temperature incubator 2, a thin-film platinum thermal resistance sensor 8, a cell culture plate 6, a 3D printing frame 7, and a resistance detection device 5. The cell culture plate 6 is placed in the constant temperature incubator 2, the cell culture plate 6 is provided with a culture hole 11, and the culture hole 11 is provided with a thin-film platinum thermal resistance sensor 8 and a 3D printing frame 7, The thin-film platinum thermal resistance sensor 8 is connected to the cell culture plate 6 through the 3D printing frame 7 ; the thin-film platinum thermal resistance sensor 8 is connected to the resistance detection device 5 .
所述细胞培养板6上设有与薄膜铂热电阻传感器8相对应的通孔10；所述3D打印框体7内设有与薄膜铂热电阻传感器8相对应的凹槽9。The cell culture plate 6 is provided with a through hole 10 corresponding to the thin-film platinum thermal resistance sensor 8 ; the 3D printing frame 7 is provided with a groove 9 corresponding to the thin-film platinum thermal resistance sensor 8 .
所述薄膜铂热电阻传感器8的数量为至少12个，所述培养孔11的数量为至少12个。The number of the thin-film platinum thermal resistance sensors 8 is at least 12, and the number of the culture wells 11 is at least 12.
还包括计算机1，所述计算机1通过无线网络终端与电阻检测装置5相连。所述无线网络终端为蓝牙4。It also includes a computer 1, which is connected to the resistance detection device 5 through a wireless network terminal. The wireless network terminal is Bluetooth 4.
还包括锂电池3，所述锂电池3设置在恒温培养箱2内，所述锂电池3与电阻检测装置5相连。It also includes a lithium battery 3, which is arranged in the constant temperature incubator 2, and the lithium battery 3 is connected with a resistance detection device 5.
所述3D打印框体7为白色光敏树脂材料通过立体光固化成型工艺打印。The 3D printing frame 7 is printed by a white photosensitive resin material through a stereolithography molding process.
本实施方式所述的无线多通道系统，使用神经干细胞作为监测对象，如图1所示，利用AutoCAD制图软件设计、立体光固化成型（SLA）工艺制作3D打印框体7，使用无毒硅酮导热胶，将鼠尾胶原包被处理后的薄膜铂热电阻传感器8（改善其生物相容性）和3D打印框体7固定在六孔细胞培养板6中，引出薄膜铂热电阻传感器8导线，在常温下使用环氧乙烷对设备进行灭菌，此步骤旨在构建细胞生长及温度传感环境。The wireless multi-channel system described in this embodiment uses neural stem cells as the monitoring object, as shown in Figure 1, uses AutoCAD drawing software design, stereolithography (SLA) process to produce 3D printing frame 7, using non-toxic silicone Thermally conductive adhesive, fix the thin-film platinum thermal resistance sensor 8 coated with rat tail collagen (to improve its biocompatibility) and the 3D printing frame 7 in the six-well cell culture plate 6, and lead out the wire of the thin-film platinum thermal resistance sensor 8 , use ethylene oxide to sterilize the device at room temperature, this step is designed to build a cell growth and temperature sensing environment.
接下来，在超净台里接种浓度为5×108/L的神经干细胞接种到薄膜铂热电阻传感器8表面，将薄膜铂热电阻传感器8与电阻检测装置5连接，放入恒温培养箱2中，开启计算机1，通过蓝牙4传输数据，实时观察其温度变化情况，总体系统效果如图2所示。Next, inoculate the neural stem cells with a concentration of 5×10 8 /L on the surface of the thin-film platinum thermal resistance sensor 8 in the ultra-clean bench, connect the thin-film platinum thermal resistance sensor 8 with the resistance detection device 5, and put it into the constant temperature incubator 2 , turn on computer 1, transmit data through Bluetooth 4, and observe its temperature changes in real time. The overall system effect is shown in Figure 2.
本实施实例设置六个实验组（神经干细胞+培养基）及六个对照组（培养基），为简洁说明，选取计算机1中的一组进行展示，图3显示接种神经干细胞的组别温度渐渐高于对照组，其温度差值逐渐稳定在0.4℃左右。图4显示实验组的细胞在贴壁过程中与对照组的温度差值变化，其在1小时内逐渐升高。In this implementation example, six experimental groups (neural stem cells + culture medium) and six control groups (medium) are set. For concise description, a group in computer 1 is selected for display. Figure 3 shows that the temperature of the group inoculated with neural stem cells is gradually increasing. Higher than the control group, the temperature difference gradually stabilized at around 0.4°C. Figure 4 shows the temperature difference between the cells in the experimental group and the control group during the attachment process, which gradually increased within 1 hour.
本实施方式能够有效的对数千数量级的细胞进行实时温度监测，同时具有体积小、响应速度快、性能稳定等优点。This embodiment can effectively monitor the temperature of thousands of cells in real time, and has the advantages of small size, fast response speed, and stable performance.
上述实施例仅为本发明的优选技术方案，而不应视为对于本发明的限制，本发明的保护范围应以权利要求记载的技术方案，包括权利要求记载的技术方案中技术特征的等同替换方案为保护范围，即在此范围内的等同替换改进，也在本发明的保护范围之内。The above-described embodiments are only preferred technical solutions of the present invention, and should not be regarded as limitations on the present invention. The protection scope of the present invention should be based on the technical solutions described in the claims, including the equivalent replacement of technical features in the technical solutions described in the claims. The solution is within the scope of protection, that is, equivalent replacement and improvement within this scope are also within the protection scope of the present invention.
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