基于人工智能的高精度菌落提取与分离系统
Artificial intelligence-enabled high-precision colony extraction and isolation system
doi: 10.37188/CO.EN-2025-0025
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摘要:
标准菌悬液在微生物诊断中具有重要意义。传统制备方法依赖人工操作,存在重复性差、效率低及生物安全隐患等问题。本研究提出一种融合大视野成像与人工智能技术的高精度自动化菌落提取分选系统,实现菌落智能筛查与定位。首先,开发了大视野成像系统,可采集90 mm培养皿高分辨图像,物理分辨率达13.2 μm,成像速度为13帧/秒;其次,应用人工智能技术实现菌落自动识别与定位,支持筛选直径1.9−2.3 mm的目标菌落;接着设计三轴运动控制平台,配合路径规划算法实现菌落高效提取,采用电动移液器进行精准菌落采集;同时开发菌悬液浓度测量模块,以650 nm激光二极管为光源,实现0.01麦氏浓度(MCF)的测量精度。最终通过大肠杆菌悬液制备验证系统性能:经17小时培养后分4次提取大肠杆菌,达到系统设定目标浓度。该工作有望实现微生物样本的快速精准制备,显著缩短检测周期,减轻医务人员工作负担。
Abstract:Standard bacterial suspensions play a crucial role in microbiological diagnosis. Traditional preparation methods, which rely heavily on manual operations, face challenges such as poor reproducibility, low efficiency, and biosafety concerns. In this study, we propose a high-precision automated colony extraction and separation system that combines large-field imaging and artificial intelligence (AI) to facilitate intelligent screening and localization of colonies. Firstly, a large-field imaging system was developed to capture high-resolution images of 90 mm Petri dishes, achieving a physical resolution of 13.2 μm and an imaging speed of 13 frames per second. Subsequently, AI technology was employed for the automatic recognition and localization of colonies, enabling the selection of target colonies with diameters ranging from 1.9 to 2.3 mm. Next, a three-axis motion control platform was designed, accompanied by a path planning algorithm for the efficient extraction of colonies. An electronic pipette was employed for accurate colony collection. Additionally, a bacterial suspension concentration measurement module was developed, incorporating a 650 nm laser diode as the light source, achieving a measurement accuracy of 0.01 McFarland concentration (MCF). Finally, the system’s performance was validated through the preparation of an E. coli suspension. After 17 hours of cultivation, E. coli was extracted four times, achieving the target concentration set by the system. This work is expected to enable rapid and accurate microbial sample preparation, significantly reducing detection cycles and alleviating the workload of healthcare personnel.
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Figure 2. High-precision automated colony extraction system. (a) Core control board. (b) Physical diagram of the optical system for colony extraction. (c) Components of the extraction system. (d) Translation transformation between the world coordinate system and the camera coordinate system. (e) Schematic diagram of dynamic solution adjustment.
Figure 3. Bacterial suspension concentration measurement system. (a) Comparison of scattering and transmission measurement values at 0.5 McFarland concentration. (b) Schematic diagram and 3D model of the bacterial suspension concentration measurement module. (c) Physical diagram of the single-wavelength measurement system. (d) Circuit diagram of the signal amplification for measurement. (e) Calibration curve of measured voltage versus standard concentration.
Figure 5. Path planning for single colony extraction. (a) 3D random path diagram. (b) This work. (c) Random path 1. (d) Random path 2. (e) Random path 3. (f) Random path 4. (g) Random path 5. (h) Comparison of Euclidean distances for different path planning strategies. (i) Host computer operation interface.
Figure 6. On-demand preparation of E. coli suspension concentration. (a) Relationship between the number of extractions and cultivation time at target concentration. (b-d) Relationship between E. coli concentration and the number of extractions after 17 hours of cultivation: (b) Group 1, (c) Group 2, (d) Group 3.
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