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工程设计学报  2022, Vol. 29 Issue (4): 510-518    DOI: 10.3785/j.issn.1006-754X.2022.00.063
整机和系统设计     
基于STM32的全自动病理染色控制系统的设计与实现
谭鑫平1(),甘浪1,林成旭1,李雪梅2,李倩2,闫旺2,陈刚2,廖广兰1,刘智勇1()
1.华中科技大学 机械科学与工程学院,湖北 武汉 430074
2.武汉康录生物技术股份有限公司,湖北 武汉 430000
Design and implementation of fully automatic pathological staining control system based on STM32
Xin-ping TAN1(),Lang GAN1,Cheng-xu LIN1,Xue-mei LI2,Qian LI2,Wang YAN2,Gang CHEN2,Guang-lan LIAO1,Zhi-yong LIU1()
1.School of Mechanical Science and Engineering, Huazhong University of Science and Technology, Wuhan 430074, China
2.Wuhan Health Care Biotechnology Co. , Ltd. , Wuhan 430000, China
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摘要:

FISH (fluorescence in situ hybridization,荧光原位杂交)染色是应用于病理分析的重要技术。传统的手工染色方式由于操作繁琐及实验条件限制难以控制杂交质量,而国内现有的染色系统存在自动化程度低等缺点。为了解决上述问题,基于STM32嵌入式系统和GUI (graphical user interface,图形用户界面)开发了一套全自动病理染色控制系统。以STM32F103ZET6作为下位机处理器,利用QT软件和C++语言设计GUI界面,实现了对多样本FISH染色过程的全自动化控制。所研发的控制系统能够满足全自动病理染色系统的控制要求,滴加试剂的位置精度达到0.05 mm,试剂体积精度达到0.6 μL,设备故障响应时间小于0.5 s,多次实验设备运行故障率小于3%,加样时试剂类型选择准确率达到100%,图像可判读率达到90%以上。该控制系统具有高效率、高鲁棒性的优点,结合全自动病理染色硬件系统进行染色实验,取得了均匀、良好的染色效果,因此具有良好的应用价值。

关键词: 全自动病理染色系统控制系统STM32F103ZET6GUI (图形用户界面)FISH (荧光原位杂交)染色技术    
Abstract:

FISH (fluorescence in situ hybridization) staining is an important technique applied to pathological analysis. Traditional manual staining method is difficult to control hybridization quality due to the cumbersome operation and the limitations of experimental conditions, while the existing domestic staining system has the disadvantages of low degree of automation. In order to solve the above problems, a fully automatic pathological staining control system was developed based on STM32 embedded system and GUI (graphical user interface). With STM32F103ZET6 as the lower processor, GUI interface designed by QT software and C++ language, the system realized the fully automatic control of multi-sample FISH staining process. The developed control system could meet the control requirements of the fully automatic pathological staining system. The position accuracy of the dropping reagent could reach 0.05 mm, the reagent volume accuracy could reach 0.6 μL, the equipment fault response time was less than 0.5 s, the failure rate of equipment in multiple experiments was less than 3%, the accuracy of reagent type selection during sample addition reached 100%, and the image legibility rate was more than 90%. The control system has the advantages of high efficiency and high robustness. Combined with the fully automatic pathological staining hardware system, the staining experiment could achieve a uniform and good staining effect. So, it has good application value.

Key words: fully automatic pathological staining system    control system    STM32F103ZET6    GUI(graphical user interface)    FISH (fluorescence in situ hybridization) staining technology
收稿日期: 2021-12-20 出版日期: 2022-09-05
CLC:  TP 273  
通讯作者: 刘智勇     E-mail: tan@hust.edu.cn;zhiyong_liu@hust.edu.cn
作者简介: 谭鑫平(1999—),男,湖南衡阳人,硕士生,从事嵌入式系统、数字图像处理和深度学习研究,E-mail:xinping_ tan@hust.edu.cnhttps://orcid.org/0000-0003-4210-3852
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引用本文:

谭鑫平,甘浪,林成旭,李雪梅,李倩,闫旺,陈刚,廖广兰,刘智勇. 基于STM32的全自动病理染色控制系统的设计与实现[J]. 工程设计学报, 2022, 29(4): 510-518.

Xin-ping TAN,Lang GAN,Cheng-xu LIN,Xue-mei LI,Qian LI,Wang YAN,Gang CHEN,Guang-lan LIAO,Zhi-yong LIU. Design and implementation of fully automatic pathological staining control system based on STM32[J]. Chinese Journal of Engineering Design, 2022, 29(4): 510-518.

链接本文:

https://www.zjujournals.com/gcsjxb/CN/10.3785/j.issn.1006-754X.2022.00.063        https://www.zjujournals.com/gcsjxb/CN/Y2022/V29/I4/510

图1  染色控制系统组成
图2  染色控制系统主电路连接示意
部件代号部件类型型号主参数精度
M1伺服电机42CME08保持转矩为0.8 Nm步距角为1.8°
M2直线电机11HY3401?25D52保持转矩为0.06 Nm步距角为1.8°
M3蠕动泵KCS?SA?A?B166流量为40 mL/min2%
M4微量注射泵Air?Z Legacy量程为200 μL±3%
M5真空泵1410V DC最终真空度为75%
M6气泵6015SE/24V DC最大持续压强为14 kPa
表1  染色控制系统主要运动部件的参数
图3  下位机模块构成
执行部件脉冲引脚电平引脚
M1PD5PD6
M2PD1PD2
M3PA1PA5
M4PF1PF2
M5PE2
M6PE0
表2  STM32 I/O模块的引脚分配
图4  下位机程序流程框图
图5  上位机模块构成
步骤操作试剂时间/min
1烤片30
2脱蜡脱蜡剂15
3洗涤100%乙醇4
4通透通透剂25
5水处理去离子水6
6酶处理蛋白酶30
7乙醇处理梯度乙醇12
8变性探针5
9杂交探针123
10洗涤洗涤液、去离子水8
11染色DAPI染色剂1
表3  面向FISH染色的全自动病理染色系统的染色步骤
图6  染色控制系统GUI界面
状态变量说明事件设定值
Pushbtn记录载玻片执行模式鼠标点击载玻片0~3(12位)
SumPushbtn记录用户是否选择载玻片没有载玻片被选择0
已有载玻片被选择1~12
Flagstart记录“开始”按钮状态按下状态1
弹起状态0
Flagstop记录“暂停”按钮状态按下状态1
弹起状态0
Permeationtime记录“通透”步骤时间编辑框输入用户设置
Proteintime记录“酶处理”步骤时间编辑框输入用户设置
Totaltime记录系统运行总时间用户设置时间实时变化
表4  界面状态变量及其事件、设定值
图7  通信内容及STM32数据传输过程
图8  多传感信息融合流程
图9  全自动病理染色系统模型渲染图及结构实物图
图10  实验所得的荧光图像
生产商仪器整机长×宽×高/mm×mm×mm

整机

质量/kg

温度

范围/℃

最低升温

速度/(℃/min)

温控精度/℃最少试剂加样量/ μL是否全自动化
中国厦门通灵生物医药科技有限公司Aliya全自动免疫组化染色仪1 075×780×870110室温~100±370
美国BioGenex公司

Xmatrx全自动病理

切片染色系统

1 163×737×1499182室温~10529±0.55
瑞士Roche公司

BenchMark ULTRA

自动病理组织染色机

1 118×841×1585295室温~10014.5
武汉康录生物技术股份有限公司笔者研发的染色系统850×500×54060室温~11029±15
表5  所研发的全自动病理染色系统与已有自动化染色系统的参数对比
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