CN117778176A - Automatic detection system and method for drug sensitivity of organoid - Google Patents
Automatic detection system and method for drug sensitivity of organoid Download PDFInfo
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Abstract
Description
技术领域：Technical areas:
本发明涉及类器官药敏检测技术领域，尤其涉及一种类器官药敏检测自动化系统及方法。The present invention relates to the technical field of organoid drug sensitivity detection, and in particular, to an automated system and method for organoid drug sensitivity detection.
背景技术：Background technique:
类器官属于三维(3D)细胞培养物，包含其代表器官的一些关键特性。此类体外培养系统包括一个自我更新干细胞群，可分化为多个器官特异性的细胞类型，与对应的器官拥有类似的空间组织并能够重现对应器官的部分功能，从而提供一个高度生理相关系统。精准医学应用中，患者衍生的类器官被证明为有价值的诊断工具。在进行治疗之前，采用从患者样本来源的类器官筛查患者体外药物反应，可为癌症和囊胞性纤维症患者的护理提供指导并预测治疗结果。Organoids are three-dimensional (3D) cell cultures that contain some of the key properties of the organs they represent. Such in vitro culture systems include a population of self-renewing stem cells that can differentiate into multiple organ-specific cell types, have a similar spatial organization to the corresponding organ, and are able to reproduce some of the functions of the corresponding organ, thereby providing a highly physiologically relevant system . Patient-derived organoids are proving to be valuable diagnostic tools in precision medicine applications. Screening patients for in vitro drug responses using organoids derived from patient samples before treatment can guide care and predict treatment outcomes for patients with cancer and cystic fibrosis.
类器官培养药敏检测主要包括类器官培养前预处理、类器官培养、类器官药敏检测等工序，其中，所述类器官药敏检测工序具体包括类器官消化、类器官接种、药物接触、药敏检测等步骤；Organoid culture drug susceptibility testing mainly includes pretreatment before organoid culture, organoid culture, organoid drug susceptibility testing and other processes. The organoid drug susceptibility testing process specifically includes organoid digestion, organoid inoculation, drug contact, Drug susceptibility testing and other steps;
以往的做法均采用纯人工手动操作整个过程的方式来完成类器官药敏检测工序，传统的通过采用纯人工手动操作的方式存在着以下问题：第一，由于类器官药敏检测工序需要专业的相关技术人员经过系统化学习后才能进行操作，从而会使得前期人员培养耗时耗力、培养学习成本比较高；第二，由于类器官药敏检测工序为精细化操作，只要有一点点操作误差就会导致实验结果完全不同，人工手动操作容易出现操作误差、实验结果无法得到保证；第三，由于类器官培养药敏检测往往需要进行大批量类器官药敏检测工序，需要重复性操作劳动，不仅效率较低，而且操作人员易疲劳。In the past, the organoid drug susceptibility testing process was completed by purely manual operation. The traditional method of purely manual operation has the following problems: First, because the organoid drug susceptibility testing process requires professional Relevant technical personnel can only operate after systematic learning, which will make the initial training of personnel time-consuming and labor-intensive, and the cost of training and learning is relatively high; secondly, because the organoid drug susceptibility testing process is a delicate operation, as long as there is a little operational error This will lead to completely different experimental results. Manual operations are prone to operational errors and the experimental results cannot be guaranteed. Thirdly, because organoid culture drug susceptibility testing often requires large batches of organoid drug susceptibility testing processes, which require repetitive operations. Not only is the efficiency low, but the operators are prone to fatigue.
发明内容：Contents of the invention:
本发明的目的就是针对现有技术存在的不足而提供一种类器官药敏检测自动化系统，能够替代以往传统的采用纯人工手动操作的方式来自动完成类器官药敏检测工序，能够大大地减少培养学习成本，类器官药敏检测工序整个操作更加精准、实验结果能够得到保证，能够减少大量的重复性操作劳动、减轻操作人员的负担，能够提升效率，能够更好地适用于各种大规模、大数据的类器官培养药敏检测；The purpose of the present invention is to provide an automated organoid drug sensitivity detection system in view of the shortcomings of the existing technology, which can replace the traditional purely manual operation method to automatically complete the organoid drug sensitivity detection process, and can greatly reduce the need for culture. With the cost of learning, the entire operation of the organoid drug susceptibility testing process is more accurate and the experimental results can be guaranteed. It can reduce a lot of repetitive operations and reduce the burden on operators, improve efficiency, and be better applicable to various large-scale, Big data-based organoid culture drug susceptibility testing;
本发明还提供了一种类器官药敏检测方法，采用多线程并发执行技术，能够更好地同时完成多组类器官药敏检测，能够更好地适用于各种大规模、大数据的类器官培养药敏检测，能够很好地适用于类器官药敏检测自动化系统。The present invention also provides an organoid drug sensitivity detection method, which adopts multi-threaded concurrent execution technology, can better complete multiple groups of organoid drug sensitivity detection at the same time, and can be better applied to various large-scale, big data organoids. Culture drug susceptibility testing can be well adapted to the organoid drug susceptibility testing automated system.
为了实现上述目的，本发明采用的技术方案是：一种类器官药敏检测自动化系统，包括任务输入模块、任务拆分模块、任务时序排列模块、类器官消化自动作业子模块、类器官接种自动作业子模块、药物接触自动作业子模块、药敏检测自动作业子模块；In order to achieve the above-mentioned purpose, the technical solution adopted by the present invention is: an automated organoid drug sensitivity detection system, including a task input module, a task splitting module, a task timing arrangement module, an organoid digestion automatic operation submodule, an organoid inoculation automatic operation submodule, a drug contact automatic operation submodule, and a drug sensitivity detection automatic operation submodule;
所述任务输入模块，用于输入、建立待执行类器官药敏检测任务；The task input module is used to input and establish organoid drug sensitivity testing tasks to be executed;
所述任务拆分模块，用于将输入、建立的待执行类器官药敏检测任务拆分为类器官消化任务、类器官接种任务、药物接触任务、药敏检测任务；The task splitting module is used to split the input and established organoid drug sensitivity testing tasks to be executed into organoid digestion tasks, organoid inoculation tasks, drug contact tasks, and drug sensitivity testing tasks;
所述任务时序排列模块，用于对拆分后的各个类器官药敏检测任务中的类器官消化任务、类器官接种任务、药物接触任务、药敏检测任务进行时序排列并生成任务时序排列表；The task timing arrangement module is used to arrange the organoid digestion tasks, organoid vaccination tasks, drug contact tasks, and drug sensitivity detection tasks in the split organoid drug sensitivity testing tasks in time series and generate a task timing arrangement table. ;
所述类器官消化自动作业子模块，用于按照所生成的任务时序排列表依次对各个类器官药敏检测任务中的类器官消化任务进行自动作业；The organoid digestion automatic operation sub-module is used to automatically operate the organoid digestion tasks in each organoid drug sensitivity detection task according to the generated task time sequence list;
所述类器官接种自动作业子模块，用于按照所生成的任务时序排列表依次对各个类器官药敏检测任务中的类器官接种任务进行自动作业；The organoid inoculation automatic operation submodule is used to automatically operate the organoid inoculation tasks in each organoid drug sensitivity detection task in sequence according to the generated task time sequence arrangement table;
所述药物接触自动作业子模块，用于按照所生成的任务时序排列表依次对各个类器官药敏检测任务中的药物接触任务进行自动作业；The drug contact automatic operation sub-module is used to automatically operate the drug contact tasks in each organoid drug sensitivity detection task according to the generated task time sequence list;
所述药敏检测自动作业子模块，用于按照所生成的任务时序排列表依次对各个类器官药敏检测任务中的药敏检测任务进行自动作业。The drug susceptibility testing automatic operation sub-module is used to automatically operate the drug susceptibility testing tasks in each organoid drug susceptibility testing task in sequence according to the generated task time sequence list.
对上述方案的进一步改进为，所述类器官消化自动作业子模块包括机械手装置、移液装置、细胞3D成像仪、离心机；A further improvement to the above solution is that the organoid digestion automatic operation sub-module includes a robot device, a pipetting device, a 3D cell imager, and a centrifuge;
所述类器官接种自动作业子模块包括机械手装置、移液装置、CO2培养箱、细胞计数仪；The organoid inoculation automatic operation sub-module includes a robot device, a pipetting device, a CO2 incubator, and a cell counter;
所述药物接触自动作业子模块包括机械手装置、移液装置、CO2培养箱；The drug contact automatic operation sub-module includes a robot device, a pipetting device, and a CO2 incubator;
所述药敏检测自动作业子模块包括机械手装置、移液装置、化学发光仪。The drug sensitivity detection automatic operation sub-module includes a manipulator device, a pipetting device, and a chemiluminescence instrument.
一种类器官药敏检测方法，适用于具有类器官消化自动作业子模块、类器官接种自动作业子模块、药物接触自动作业子模块、药敏检测自动作业子模块的类器官药敏检测自动化系统，包括以下步骤：An organoid drug susceptibility testing method, suitable for an organoid drug susceptibility testing automated system having an organoid digestion automatic operation submodule, an organoid inoculation automatic operation submodule, a drug contact automatic operation submodule, and a drug sensitivity detection automatic operation submodule, Includes the following steps:
A、对各个待执行类器官药敏检测任务进行输入、建立；A. Input and establish each organoid drug sensitivity testing task to be executed;
B、将输入、建立的待执行类器官药敏检测任务拆分为类器官消化任务、类器官接种任务、药物接触任务、药敏检测任务；B. Split the input and established organoid drug susceptibility testing tasks to be executed into organoid digestion tasks, organoid inoculation tasks, drug contact tasks, and drug susceptibility testing tasks;
C、将拆分后的各个类器官药敏检测任务中的类器官消化任务、类器官接种任务、药物接触任务、药敏检测任务进行时序排列并生成任务时序排列表；C. Arrange the organoid digestion task, organoid inoculation task, drug contact task, and drug sensitivity detection task in each split organoid drug sensitivity detection task in time sequence and generate a task time sequence arrangement table;
D、类器官消化自动作业子模块进行自动作业：所述类器官消化自动作业子模块，用于按照所生成的任务时序排列表依次对各个类器官药敏检测任务中的类器官消化任务进行自动作业；D. The organoid digestion automatic operation sub-module performs automatic operations: the organoid digestion automatic operation sub-module is used to automatically perform organoid digestion tasks in each organoid drug sensitivity detection task according to the generated task time sequence list. Operation;
类器官接种自动作业子模块进行自动作业：所述类器官接种自动作业子模块，用于按照所生成的任务时序排列表依次对各个类器官药敏检测任务中的类器官接种任务进行自动作业；The organoid inoculation automatic operation submodule performs automatic operation: the organoid inoculation automatic operation submodule is used to automatically operate the organoid inoculation tasks in each organoid drug sensitivity detection task in sequence according to the generated task time sequence arrangement table;
药物接触自动作业子模块进行自动作业：所述药物接触自动作业子模块，用于按照所生成的任务时序排列表依次对各个类器官药敏检测任务中的药物接触任务进行自动作业；The drug contact automatic operation submodule performs automatic operation: the drug contact automatic operation submodule is used to automatically operate the drug contact tasks in each organoid drug sensitivity detection task in sequence according to the generated task time sequence arrangement table;
药敏检测自动作业子模块进行自动作业：所述药敏检测自动作业子模块，用于按照所生成的任务时序排列表依次对各个类器官药敏检测任务中的药敏检测任务进行自动作业。The drug sensitivity detection automatic operation sub-module performs automatic operations: the drug sensitivity detection automatic operation sub-module is used to automatically operate the drug sensitivity detection tasks in each organoid drug sensitivity detection task in sequence according to the generated task time sequence list.
对上述方案的进一步改进为，所述类器官消化自动作业子模块包括机械手装置、移液装置、细胞3D成像仪、离心机；A further improvement to the above solution is that the organoid digestion automatic operation sub-module includes a robot device, a pipetting device, a 3D cell imager, and a centrifuge;
所述步骤D中的类器官消化自动作业子模块进行自动作业具体包括以下步骤：The automatic operation of the organoid digestion automatic operation sub-module in step D specifically includes the following steps:
通过移液装置向培养皿中加入凝胶消化液；Add gel digestion solution to the Petri dish using a pipetting device;
通过细胞3D成像仪进行观察；Observed through cell 3D imager;
待类器官消化成均一的含3-5个细胞的细胞团时，通过移液装置加入HBSS终止消化；When the organoids are digested into a uniform cell mass containing 3-5 cells, add HBSS through a pipetting device to terminate the digestion;
通过机械手装置将终止消化后的细胞团送入离心机进行离心，弃上清；Send the digested cell pellets to a centrifuge through a robotic device for centrifugation, and discard the supernatant;
离心后，通过移液装置加入HBSS重悬；After centrifugation, add HBSS through a pipetting device to resuspend;
通过机械手装置将重悬后的细胞团送入离心机进行离心，弃上清。The resuspended cell pellet was sent to a centrifuge by a robot device for centrifugation, and the supernatant was discarded.
对上述方案的进一步改进为，所述类器官接种自动作业子模块包括机械手装置、移液装置、CO2培养箱、细胞计数仪；A further improvement to the above solution is that the organoid inoculation automatic operation sub-module includes a manipulator device, a pipetting device, a CO 2 incubator, and a cell counter;
所述步骤D中的类器官接种自动作业子模块进行自动作业具体包括以下步骤：The automatic operation of the organoid inoculation automatic operation submodule in step D specifically includes the following steps:
通过机械手装置、移液装置、细胞计数仪的配合，进行类器官接种；Organoid inoculation is carried out through the cooperation of a robot device, a pipetting device, and a cell counter;
通过机械手装置、移液装置的配合设置阴性对照组；A negative control group was set up by cooperating with a manipulator device and a pipetting device;
通过机械手装置、移液装置的配合设置空白对照组。A blank control group was set up through the cooperation of the robot device and the pipetting device.
对上述方案的进一步改进为，所述类器官接种具体包括以下操作：A further improvement to the above scheme is that the organoid inoculation specifically includes the following operations:
用癌种培养基重悬后进行细胞计数；Resuspend in cancer culture medium and count cells;
所述设置阴性对照组具体包括以下操作：The setting of the negative control group specifically includes the following operations:
将细胞悬液与生物支架按比例进行充分稀释混匀后，将混合胶分液到微孔板中；After the cell suspension and biological scaffold are fully diluted and mixed in proportion, the mixed gel is dispensed into a microwell plate;
所述设置阴性对照组具体包括以下操作：The setting of the negative control group specifically includes the following operations:
将对应癌种培养基与生物支架按比例进行充分稀释混匀后，将混合胶分液到微孔板中。After fully diluting and mixing the corresponding cancer culture medium and biological scaffold in proportion, the mixed gel is dispensed into a microwell plate.
对上述方案的进一步改进为，所述药物接触自动作业子模块包括机械手装置、移液装置、CO2培养箱；A further improvement to the above solution is that the drug contact automatic operation sub-module includes a manipulator device, a pipetting device, and a CO 2 incubator;
通过机械手装置、移液装置的配合对类器官接种中所设置的药物测试组进行添加药液；The drug solution is added to the drug test group set up in the organoid inoculation through the cooperation of the robot device and the pipetting device;
通过机械手装置、移液装置的配合对类器官接种中所设置的药阴性对照组进行添加培养基；Through the cooperation of the robot device and the pipetting device, the culture medium is added to the drug negative control group set up in the organoid inoculation;
通过机械手装置、移液装置的配合对类器官接种中所设置的空白对照组进行添加培养基；Add culture medium to the blank control group set during organoid inoculation through the cooperation of the robot device and the pipetting device;
通过机械手装置将加完药的微孔板送入CO2培养箱进行培养。The microplate with the medicine added is sent to the CO 2 incubator through the robot device for culture.
对上述方案的进一步改进为，所述药物测试组接种有类器官、添加有药液；A further improvement to the above scheme is that the drug test group is inoculated with organoids and added with medicinal liquid;
所述阴性对照组接种有类器官、不添加药物、添加有培养基；The negative control group was inoculated with organoids, no drugs, and medium;
所述空白对照组不含类器官、不添加药物、添加有培养基；The blank control group does not contain organoids, no drugs, and media;
将加完药的微孔板送入CO2培养箱内进行培养。Put the microplate with medicine added into a CO 2 incubator for culture.
对上述方案的进一步改进为，所述药敏检测自动作业子模块包括机械手装置、移液装置、化学发光仪；A further improvement to the above scheme is that the automatic operation submodule of drug sensitivity detection includes a manipulator device, a liquid transfer device, and a chemiluminescence instrument;
所述步骤D中的药敏检测自动作业子模块进行自动作业具体包括以下步骤：The automatic operation of the drug sensitivity detection automatic operation sub-module in step D specifically includes the following steps:
通过机械手装置、移液装置的配合分别对完成药物接触后的药物测试组、阴性对照组、空白对照组添加发光试剂；Through the cooperation of the manipulator device and the pipetting device, luminescent reagents are added to the drug test group, negative control group, and blank control group after drug contact;
通过机械手装置将添加完发光试剂的药物测试组、阴性对照组、空白对照组送入化学发光仪进行化学发光检测。The drug test group, negative control group, and blank control group that have added luminescent reagents are sent to the chemiluminescence instrument through the manipulator device for chemiluminescence detection.
本发明有益效果在于：本发明提供的一种类器官药敏检测自动化系统，包括任务输入模块、任务拆分模块、任务时序排列模块、类器官消化自动作业子模块、类器官接种自动作业子模块、药物接触自动作业子模块、药敏检测自动作业子模块；The beneficial effects of the present invention are: the invention provides an automated organoid drug sensitivity detection system, which includes a task input module, a task splitting module, a task sequence arrangement module, an automatic operation sub-module for organoid digestion, and an automatic operation sub-module for organoid inoculation. Drug contact automatic operation sub-module, drug sensitivity detection automatic operation sub-module;
所述任务输入模块，用于输入、建立待执行类器官药敏检测任务；The task input module is used to input and establish the organoid drug sensitivity detection task to be executed;
所述任务拆分模块，用于将输入、建立的待执行类器官药敏检测任务拆分为类器官消化任务、类器官接种任务、药物接触任务、药敏检测任务；The task splitting module is used to split the input and established organoid drug sensitivity testing tasks to be executed into organoid digestion tasks, organoid inoculation tasks, drug contact tasks, and drug sensitivity testing tasks;
所述任务时序排列模块，用于对拆分后的各个类器官药敏检测任务中的类器官消化任务、类器官接种任务、药物接触任务、药敏检测任务进行时序排列并生成任务时序排列表；The task timing arrangement module is used to arrange the organoid digestion tasks, organoid vaccination tasks, drug contact tasks, and drug sensitivity detection tasks in the split organoid drug sensitivity testing tasks in time series and generate a task timing arrangement table. ;
所述类器官消化自动作业子模块，用于按照所生成的任务时序排列表依次对各个类器官药敏检测任务中的类器官消化任务进行自动作业；The organoid digestion automatic operation submodule is used to automatically operate the organoid digestion tasks in each organoid drug sensitivity detection task in sequence according to the generated task time sequence arrangement table;
所述类器官接种自动作业子模块，用于按照所生成的任务时序排列表依次对各个类器官药敏检测任务中的类器官接种任务进行自动作业；The organoid inoculation automatic operation submodule is used to automatically operate the organoid inoculation tasks in each organoid drug sensitivity detection task in sequence according to the generated task time sequence arrangement table;
所述药物接触自动作业子模块，用于按照所生成的任务时序排列表依次对各个类器官药敏检测任务中的药物接触任务进行自动作业；The drug contact automatic operation sub-module is used to automatically operate the drug contact tasks in each organoid drug sensitivity detection task according to the generated task time sequence list;
所述药敏检测自动作业子模块，用于按照所生成的任务时序排列表依次对各个类器官药敏检测任务中的药敏检测任务进行自动作业；The drug susceptibility testing automatic operation sub-module is used to automatically operate the drug susceptibility testing tasks in each organoid drug susceptibility testing task in sequence according to the generated task time sequence list;
相较于以往传统的采用纯人工手动操作的方式来完成类器官药敏检测工序，采用本发明的一种类器官药敏检测自动化系统来进行类器官药敏检测工序时，具有以下优点：Compared with the traditional use of pure manual operation to complete the organoid drug sensitivity testing process, using an organoid drug sensitivity testing automated system of the present invention to perform the organoid drug sensitivity testing process has the following advantages:
1、相比以往需要相关技术人员经过漫长的系统化学习才能进行类器官药敏检测工序，采用本发明的一种类器官培养自动化系统来进行类器官药敏检测工序时，整个需要手动操作的过程较少，较容易上手，能够大大地减少培养学习成本；1. Compared with the past, which required relevant technicians to go through lengthy systematic learning to carry out the organoid drug sensitivity testing process, when using an organoid culture automated system of the present invention to carry out the organoid drug sensitivity testing process, the entire process requires manual operation. Less, easier to use, and can greatly reduce training and learning costs;
2、相比人为手动操作，容易出现操作误差，采用本发明的一种类器官药敏检测自动化系统来进行类器官药敏检测工序时，通过结合自动化设备实现自动化操作，不易出错，实验整个操作更加精准、实验结果能够得到保证；2. Compared with manual operation, operational errors are prone to occur. When using an organoid drug sensitivity detection automated system of the present invention to carry out the organoid drug sensitivity detection process, the automated operation is realized by combining automated equipment, which is less prone to errors and the entire experimental operation is more accurate. Accuracy and experimental results can be guaranteed;
3、相比人为手动操作，需要重复性操作劳动，特别是一些大规模、大数据的类器官培养药敏检测，需要反反复复重复非常多次的操作，采用本发明的一种类器官药敏检测自动化系统来进行类器官药敏检测工序时，不仅能够减少大量的重复性操作劳动、减轻操作人员的负担，而且能够提升效率；3. Compared with manual operation, which requires repetitive operation labor, especially some large-scale, big data organoid culture drug sensitivity detection, which requires repeated operations many times, the use of an organoid drug sensitivity detection automation system of the present invention to perform organoid drug sensitivity detection process can not only reduce a large amount of repetitive operation labor and reduce the burden on operators, but also improve efficiency;
本发明的一种类器官药敏检测自动化系统能够更好地适用于各种大规模、大数据的类器官培养药敏检测；The automated organoid drug susceptibility testing system of the present invention can be better adapted to various large-scale, big data organoid culture drug susceptibility testing;
由于类器官药敏检测工序需要一个较长的周期，如果每次都仅对其中一个样本进行类器官药敏检测工序，待完成当前样本的类器官药敏检测工序后才对下一样本进行类器官药敏检测工序，整体操作不现实；本发明的一种类器官药敏检测方法，先将各个待执行类器官药敏检测任务进行输入、建立，将输入、建立的待执行类器官药敏检测任务拆分为类器官消化任务、类器官接种任务、药物接触任务、药敏检测任务，通过类器官消化自动作业子模块、类器官接种自动作业子模块、药物接触自动作业子模块、药敏检测自动作业子模块分别对相对应的任务进行自动化执行，本发明的一种类器官药敏检测方法通过采用多线程并发执行技术，同时对多个待执行类器官药敏检测任务进行并发执行，能够充分地调动各个自动作业子模块同步进行运转，能够更好地同时完成多组类器官药敏检测，能够更好地适用于各种大规模、大数据的类器官培养药敏检测，能够很好地适用于类器官药敏检测自动化系统。Since the organoid drug susceptibility testing process requires a long cycle, if only one sample is subjected to the organoid drug susceptibility testing process at a time, the organoid drug susceptibility testing process of the current sample will be completed before the organoid drug susceptibility testing process is completed on the next sample. The overall operation of the organ drug susceptibility testing process is unrealistic; the organoid drug susceptibility testing method of the present invention first inputs and establishes each organoid drug susceptibility testing task to be performed, and then inputs and establishes the organoid drug susceptibility testing to be performed. The tasks are divided into organoid digestion tasks, organoid vaccination tasks, drug contact tasks, and drug sensitivity testing tasks. Through the organoid digestion automatic operation sub-module, organoid vaccination automatic operation sub-module, drug contact automatic operation sub-module, and drug sensitivity testing The automatic operation sub-module automatically executes corresponding tasks respectively. An organoid drug sensitivity detection method of the present invention adopts multi-threaded concurrent execution technology to concurrently execute multiple organoid drug sensitivity detection tasks to be executed at the same time, which can fully It can effectively mobilize each automatic operation sub-module to run synchronously, which can better complete multiple sets of organoid drug susceptibility testing at the same time, and can be better applied to various large-scale, big data organoid culture drug susceptibility testing. Suitable for automated system for organoid drug susceptibility testing.
附图说明：Picture description:
图1为本发明的一种类器官药敏检测自动化系统的结构示意图。Figure 1 is a schematic structural diagram of an automated system for organoid drug sensitivity detection according to the present invention.
图2为本发明的一种类器官药敏检测方法的流程示意图。Figure 2 is a schematic flow chart of an organoid drug sensitivity detection method of the present invention.
图3为本发明一种类器官药敏检测自动化系统工作时长与人工工作时长的对比图。Figure 3 is a comparison chart of the working hours of an automated system for organoid drug sensitivity detection according to the present invention and the manual working hours.
图4为本发明一种类器官药敏检测自动化系统药敏检测流程与人工实验的稳定性对比图。Figure 4 is a stability comparison diagram between the drug sensitivity detection process of an automated organoid drug sensitivity detection system and manual experiments according to the present invention.
具体实施方式：Detailed ways:
下面结合附图对本发明作进一步的说明，如图1所示，本发明的一种类器官药敏检测自动化系统，包括任务输入模块、任务拆分模块、任务时序排列模块、类器官消化自动作业子模块、类器官接种自动作业子模块、药物接触自动作业子模块、药敏检测自动作业子模块；The present invention will be further described below with reference to the accompanying drawings. As shown in Figure 1, an automated organoid drug sensitivity detection system of the present invention includes a task input module, a task splitting module, a task sequence arrangement module, and an organoid digestion automatic operation sub-system. Module, automatic operation sub-module for organoid vaccination, automatic operation sub-module for drug exposure, and automatic operation sub-module for drug susceptibility testing;
所述任务输入模块，用于输入、建立待执行类器官药敏检测任务；The task input module is used to input and establish organoid drug sensitivity testing tasks to be executed;
所述任务拆分模块，用于将输入、建立的待执行类器官药敏检测任务拆分为类器官消化任务、类器官接种任务、药物接触任务、药敏检测任务；The task splitting module is used to split the input and established organoid drug sensitivity testing tasks to be executed into organoid digestion tasks, organoid inoculation tasks, drug contact tasks, and drug sensitivity testing tasks;
所述任务时序排列模块，用于对拆分后的各个类器官药敏检测任务中的类器官消化任务、类器官接种任务、药物接触任务、药敏检测任务进行时序排列并生成任务时序排列表；The task timing arrangement module is used to arrange the organoid digestion tasks, organoid vaccination tasks, drug contact tasks, and drug sensitivity detection tasks in the split organoid drug sensitivity testing tasks in time series and generate a task timing arrangement table. ;
所述类器官消化自动作业子模块，用于按照所生成的任务时序排列表依次对各个类器官药敏检测任务中的类器官消化任务进行自动作业；The organoid digestion automatic operation sub-module is used to automatically operate the organoid digestion tasks in each organoid drug sensitivity detection task according to the generated task time sequence list;
所述类器官接种自动作业子模块，用于按照所生成的任务时序排列表依次对各个类器官药敏检测任务中的类器官接种任务进行自动作业；The organoid inoculation automatic operation sub-module is used to automatically operate the organoid inoculation tasks in each organoid drug sensitivity testing task according to the generated task time sequence list;
所述药物接触自动作业子模块，用于按照所生成的任务时序排列表依次对各个类器官药敏检测任务中的药物接触任务进行自动作业；The drug contact automatic operation sub-module is used to automatically operate the drug contact tasks in each organoid drug sensitivity detection task according to the generated task time sequence list;
所述药敏检测自动作业子模块，用于按照所生成的任务时序排列表依次对各个类器官药敏检测任务中的药敏检测任务进行自动作业。The drug susceptibility testing automatic operation sub-module is used to automatically operate the drug susceptibility testing tasks in each organoid drug susceptibility testing task in sequence according to the generated task time sequence list.
所述类器官消化自动作业子模块包括机械手装置、移液装置、细胞3D成像仪、离心机；The organoid digestion automatic operation sub-module includes a robot device, a pipetting device, a 3D cell imager, and a centrifuge;
所述类器官接种自动作业子模块包括机械手装置、移液装置、CO2培养箱、细胞计数仪；The organoid inoculation automatic operation sub-module includes a robot device, a pipetting device, a CO2 incubator, and a cell counter;
所述药物接触自动作业子模块包括机械手装置、移液装置、CO2培养箱；The drug contact automatic operation sub-module includes a robot device, a pipetting device, and a CO2 incubator;
所述药敏检测自动作业子模块包括机械手装置、移液装置、化学发光仪；其中，各个自动作业子模块中所包含的部件之间可以为同一结构，也可以为相独立设置的结构，如类器官消化自动作业子模块中的机械手装置与类器官接种自动作业子模块中的机械手装置既可以为同一结构，也可以为相独立设置的结构。The automatic operation submodule for drug sensitivity detection includes a robotic arm device, a pipetting device, and a chemiluminometer; wherein the components contained in each automatic operation submodule can be of the same structure or of independently arranged structures, such as the robotic arm device in the automatic operation submodule for organoid digestion and the robotic arm device in the automatic operation submodule for organoid inoculation can be of the same structure or of independently arranged structures.
相较于以往传统的采用纯人工手动操作的方式来完成类器官药敏检测工序，采用本发明的一种类器官药敏检测自动化系统来进行类器官药敏检测工序时，具有以下优点：Compared with the traditional use of pure manual operation to complete the organoid drug sensitivity testing process, using an organoid drug sensitivity testing automated system of the present invention to perform the organoid drug sensitivity testing process has the following advantages:
1、相比以往需要相关技术人员经过漫长的系统化学习才能进行类器官药敏检测工序，采用本发明的一种类器官培养自动化系统来进行类器官药敏检测工序时，整个需要手动操作的过程较少，较容易上手，能够大大地减少培养学习成本；1. Compared with the past, which required relevant technicians to go through lengthy systematic learning to carry out the organoid drug sensitivity testing process, when using an organoid culture automated system of the present invention to carry out the organoid drug sensitivity testing process, the entire process requires manual operation. Less, easier to use, and can greatly reduce training and learning costs;
2、相比人为手动操作，容易出现操作误差，采用本发明的一种类器官药敏检测自动化系统来进行类器官药敏检测工序时，通过结合自动化设备实现自动化操作，不易出错，实验整个操作更加精准、实验结果能够得到保证；2. Compared with manual operation, operational errors are prone to occur. When using an organoid drug sensitivity detection automated system of the present invention to perform the organoid drug sensitivity detection process, the automated operation is realized by combining automated equipment, which is less prone to errors and the entire experimental operation is more accurate. Accuracy and experimental results can be guaranteed;
3、相比人为手动操作，需要重复性操作劳动，特别是一些大规模、大数据的类器官培养药敏检测，需要反反复复重复非常多次的操作，采用本发明的一种类器官药敏检测自动化系统来进行类器官药敏检测工序时，不仅能够减少大量的重复性操作劳动、减轻操作人员的负担，而且能够提升效率；3. Compared with manual operation, which requires repetitive operation labor, especially some large-scale, big data organoid culture drug susceptibility testing, which requires repeated operations many times, using an organoid drug susceptibility test of the present invention When the detection automation system performs the organoid drug susceptibility testing process, it can not only reduce a large amount of repetitive operations and reduce the burden on operators, but also improve efficiency;
本发明的一种类器官药敏检测自动化系统能够更好地适用于各种大规模、大数据的类器官培养药敏检测。The automated organoid drug susceptibility testing system of the present invention can be better adapted to various large-scale, big data organoid culture drug susceptibility testing.
如图2所示，本发明的一种类器官药敏检测方法，适用于具有类器官消化自动作业子模块、类器官接种自动作业子模块、药物接触自动作业子模块、药敏检测自动作业子模块的类器官药敏检测自动化系统，包括以下步骤：As shown in Figure 2, the organoid drug sensitivity detection method of the present invention is suitable for use with an automatic operation sub-module for organoid digestion, an automatic operation sub-module for organoid inoculation, an automatic operation sub-module for drug contact, and an automatic operation sub-module for drug sensitivity detection. The automated system for organoid drug susceptibility testing includes the following steps:
A、对各个待执行类器官药敏检测任务进行输入、建立；A. Input and establish each organoid drug sensitivity testing task to be executed;
B、将输入、建立的待执行类器官药敏检测任务拆分为类器官消化任务、类器官接种任务、药物接触任务、药敏检测任务；B. Split the input and established organoid drug susceptibility testing tasks to be executed into organoid digestion tasks, organoid inoculation tasks, drug contact tasks, and drug susceptibility testing tasks;
C、将拆分后的各个类器官药敏检测任务中的类器官消化任务、类器官接种任务、药物接触任务、药敏检测任务进行时序排列并生成任务时序排列表；C. Arrange the organoid digestion tasks, organoid inoculation tasks, drug contact tasks, and drug susceptibility testing tasks in time series among the split organoid drug sensitivity testing tasks and generate a task time series list;
D、类器官消化自动作业子模块进行自动作业：所述类器官消化自动作业子模块，用于按照所生成的任务时序排列表依次对各个类器官药敏检测任务中的类器官消化任务进行自动作业；D. The organoid digestion automatic operation sub-module performs automatic operations: the organoid digestion automatic operation sub-module is used to automatically perform organoid digestion tasks in each organoid drug sensitivity detection task according to the generated task time sequence list. Operation;
类器官接种自动作业子模块进行自动作业：所述类器官接种自动作业子模块，用于按照所生成的任务时序排列表依次对各个类器官药敏检测任务中的类器官接种任务进行自动作业；The organoid inoculation automatic operation submodule performs automatic operation: the organoid inoculation automatic operation submodule is used to automatically operate the organoid inoculation tasks in each organoid drug sensitivity detection task in sequence according to the generated task time sequence arrangement table;
药物接触自动作业子模块进行自动作业：所述药物接触自动作业子模块，用于按照所生成的任务时序排列表依次对各个类器官药敏检测任务中的药物接触任务进行自动作业；The drug contact automatic operation submodule performs automatic operation: the drug contact automatic operation submodule is used to automatically operate the drug contact tasks in each organoid drug sensitivity detection task in sequence according to the generated task time sequence arrangement table;
药敏检测自动作业子模块进行自动作业：所述药敏检测自动作业子模块，用于按照所生成的任务时序排列表依次对各个类器官药敏检测任务中的药敏检测任务进行自动作业。The drug sensitivity detection automatic operation sub-module performs automatic operations: the drug sensitivity detection automatic operation sub-module is used to automatically operate the drug sensitivity detection tasks in each organoid drug sensitivity detection task in sequence according to the generated task time sequence list.
由于类器官药敏检测工序需要一个较长的周期，如果每次都仅对其中一个样本进行类器官药敏检测工序，待完成当前样本的类器官药敏检测工序后才对下一样本进行类器官药敏检测工序，整体操作不现实；本发明的一种类器官药敏检测方法，先将各个待执行类器官药敏检测任务进行输入、建立，将输入、建立的待执行类器官药敏检测任务拆分为类器官消化任务、类器官接种任务、药物接触任务、药敏检测任务，通过类器官消化自动作业子模块、类器官接种自动作业子模块、药物接触自动作业子模块、药敏检测自动作业子模块分别对相对应的任务进行自动化执行，本发明的一种类器官药敏检测方法通过采用多线程并发执行技术，同时对多个待执行类器官药敏检测任务进行并发执行，能够充分地调动各个自动作业子模块同步进行运转，能够更好地同时完成多组类器官药敏检测，能够更好地适用于各种大规模、大数据的类器官培养药敏检测，能够很好地适用于类器官药敏检测自动化系统。Since the organoid drug susceptibility testing process requires a long cycle, if only one sample is subjected to the organoid drug susceptibility testing process at a time, the organoid drug susceptibility testing process of the current sample will be completed before the organoid drug susceptibility testing process is completed on the next sample. The overall operation of the organ drug susceptibility testing process is unrealistic; the organoid drug susceptibility testing method of the present invention first inputs and establishes each organoid drug susceptibility testing task to be performed, and then inputs and establishes the organoid drug susceptibility testing to be performed. The tasks are divided into organoid digestion tasks, organoid vaccination tasks, drug contact tasks, and drug sensitivity testing tasks. Through the organoid digestion automatic operation sub-module, organoid vaccination automatic operation sub-module, drug contact automatic operation sub-module, and drug sensitivity testing The automatic operation sub-module automatically executes corresponding tasks respectively. An organoid drug sensitivity detection method of the present invention adopts multi-threaded concurrent execution technology to concurrently execute multiple organoid drug sensitivity detection tasks to be executed at the same time, which can fully It can effectively mobilize each automatic operation sub-module to run synchronously, which can better complete multiple sets of organoid drug susceptibility testing at the same time, and can be better applied to various large-scale, big data organoid culture drug susceptibility testing. Suitable for automated system for organoid drug susceptibility testing.
所述类器官消化自动作业子模块包括机械手装置、移液装置、细胞3D成像仪、离心机；The organoid digestion automatic operation sub-module includes a robot device, a pipetting device, a 3D cell imager, and a centrifuge;
所述步骤D中的类器官消化自动作业子模块进行自动作业具体包括以下步骤：The automatic operation of the organoid digestion automatic operation sub-module in step D specifically includes the following steps:
通过移液装置向培养皿中加入凝胶消化液；Add gel digestion solution to the Petri dish using a pipetting device;
通过细胞3D成像仪进行观察；Observed through cell 3D imager;
待类器官消化成均一的含3-5个细胞的细胞团时，通过移液装置加入HBSS终止消化；When the organoids are digested into a uniform cell mass containing 3-5 cells, add HBSS through a pipetting device to terminate the digestion;
通过机械手装置将终止消化后的细胞团送入离心机进行离心，弃上清；Send the digested cell pellets to a centrifuge through a robotic device for centrifugation, and discard the supernatant;
离心后，通过移液装置加入HBSS重悬；After centrifugation, add HBSS via pipetting device to resuspend;
通过机械手装置将重悬后的细胞团送入离心机进行离心，弃上清。Send the resuspended cell pellet to the centrifuge for centrifugation through a robot device, and discard the supernatant.
所述用于终止消化的HBSS的量为凝胶消化液体积的1.5倍；The amount of HBSS used to terminate the digestion is 1.5 times the volume of the gel digestion solution;
在将终止消化后的细胞团送入离心机进行离心时，以1200rpm离心3min；When sending the digested cell pellets into a centrifuge for centrifugation, centrifuge at 1200 rpm for 3 minutes;
在将重悬后的细胞团送入离心机进行离心时，以1200rpm离心3min。When the resuspended cell pellet is sent to the centrifuge for centrifugation, centrifuge at 1200 rpm for 3 minutes.
相较于以往传统通过人工手动的方式进行类器官消化，本发明通过机械手装置、移液装置、细胞3D成像仪、离心机的结合，能够替代人手操作并实现自动类器官消化。Compared with the traditional manual digestion of organoids in the past, the present invention can replace manual operation and realize automatic organoid digestion through the combination of a robot device, a pipetting device, a 3D cell imager, and a centrifuge.
所述类器官接种自动作业子模块包括机械手装置、移液装置、CO2培养箱、细胞计数仪；The organoid inoculation automatic operation sub-module includes a robot device, a pipetting device, a CO2 incubator, and a cell counter;
所述步骤D中的类器官接种自动作业子模块进行自动作业具体包括以下步骤：The automatic operation of the organoid inoculation automatic operation submodule in step D specifically includes the following steps:
通过机械手装置、移液装置、细胞计数仪的配合，以每孔2000活细胞进行类器官接种；Through the cooperation of robot device, pipetting device and cell counter, organoid inoculation is carried out with 2000 viable cells per well;
通过机械手装置、移液装置的配合设置阴性对照组；A negative control group is set up through the cooperation of the robot device and the pipetting device;
通过机械手装置、移液装置的配合设置空白对照组。A blank control group was set up through the cooperation of the robot device and the pipetting device.
所述类器官接种具体包括以下操作：The organoid inoculation specifically includes the following operations:
用200μl对应癌种培养基重悬后进行细胞计数；Resuspend in 200 μl of the corresponding cancer medium and count the cells;
所述设置阴性对照组具体包括以下操作：The setting of the negative control group specifically includes the following operations:
将细胞悬液与生物支架按1:1.3的比例进行充分稀释混匀后，将混合胶以10μl/孔分液到384微孔板中，其中，阴性对照组的孔数为12；其中，所述生物支架可以为生物基质、基底膜、基质胶、胶原、明胶、细胞外骨架、水凝胶或化学胶。After the cell suspension and biological scaffold are fully diluted and mixed at a ratio of 1:1.3, the mixed gel is dispensed into a 384 microwell plate at 10 μl/well. Among them, the number of holes in the negative control group is 12; among them, the The biological scaffold can be a biological matrix, basement membrane, Matrigel, collagen, gelatin, extracellular skeleton, hydrogel or chemical glue.
所述设置阴性对照组具体包括以下操作：The setting of the negative control group specifically includes the following operations:
将对应癌种培养基与生物支架按1:1.3的比例进行充分稀释混匀后，将混合胶以10μl/孔分液到384微孔板中，其中，空白组的孔数为6。After fully diluting and mixing the corresponding cancer culture medium and biological scaffold at a ratio of 1:1.3, dispense the mixed gel into a 384 microwell plate at 10 μl/well, in which the number of holes in the blank group is 6.
相较于以往传统通过人工手动的方式进行类器官接种，本发明通过机械手装置、移液装置、CO2培养箱、细胞计数仪的结合，能够替代人手操作并实现自动类器官接种。Compared with the traditional manual method of organoid inoculation, the present invention can replace manual operation and realize automatic organoid inoculation through the combination of a robotic device, a pipetting device, a CO2 incubator, and a cell counter.
所述药物接触自动作业子模块包括机械手装置、移液装置、CO2培养箱；The drug contact automatic operation sub-module includes a robot device, a pipetting device, and a CO 2 incubator;
通过机械手装置、移液装置的配合对类器官接种中所设置的药物测试组进行添加药液；The drug solution is added to the drug test group set up in the organoid inoculation through the cooperation of the robot device and the pipetting device;
通过机械手装置、移液装置的配合对类器官接种中所设置的药阴性对照组进行添加培养基；Through the cooperation of the robot device and the pipetting device, the culture medium is added to the drug negative control group set up in the organoid inoculation;
通过机械手装置、移液装置的配合对类器官接种中所设置的空白对照组进行添加培养基；Add culture medium to the blank control group set during organoid inoculation through the cooperation of the robot device and the pipetting device;
通过机械手装置将加完药的微孔板送入CO2培养箱进行培养。The microplate with added drugs is sent into the CO2 incubator for cultivation by a robotic arm device.
所述药物测试组接种有类器官、添加有药液6μl；The drug test group was inoculated with organoids and added with 6 μl of medicinal solution;
所述阴性对照组接种有类器官、不添加药物、添加有培养基6μl；The negative control group was inoculated with organoids, no drugs, and 6 μl of culture medium;
所述空白对照组不含类器官、不添加药物、添加有培养基6μl；The blank control group did not contain organoids, no drugs, and 6 μl of culture medium;
将加完药的微孔板送入CO2培养箱内在37℃、5％的CO2浓度条件下进行培养4天。Put the microplate with medicine added into a CO 2 incubator and culture it for 4 days at 37°C and 5% CO 2 concentration.
相较于以往传统通过人工手动的方式进行药物接触，本发明通过机械手装置、移液装置、CO2培养箱的结合，能够替代人手操作并实现自动药物接触。Compared with the traditional manual method of drug contact, the present invention can replace manual operation and realize automatic drug contact through the combination of a robot device, a pipetting device, and a CO2 incubator.
所述药敏检测自动作业子模块包括机械手装置、移液装置、化学发光仪；The drug sensitivity detection automatic operation sub-module includes a manipulator device, a pipetting device, and a chemiluminescence instrument;
所述步骤D中的药敏检测自动作业子模块进行自动作业具体包括以下步骤：The automatic operation of the drug sensitivity detection automatic operation sub-module in step D specifically includes the following steps:
通过机械手装置、移液装置的配合分别对完成药物接触后的药物测试组、阴性对照组、空白对照组添加发光试剂；Through the cooperation of the manipulator device and the pipetting device, luminescent reagents are added to the drug test group, negative control group, and blank control group after drug contact;
通过机械手装置将添加完发光试剂的药物测试组、阴性对照组、空白对照组送入化学发光仪进行化学发光检测。The drug test group, negative control group, and blank control group that have added luminescent reagents are sent to the chemiluminescence instrument through the manipulator device for chemiluminescence detection.
相较于以往传统通过人工手动的方式进行药敏检测，本发明通过机械手装置、移液装置、化学发光仪的结合，能够替代人手操作并实现自动药敏检测。Compared with the traditional manual method of conducting drug susceptibility testing in the past, the present invention can replace human operation and realize automatic drug susceptibility testing through the combination of a manipulator device, a pipetting device, and a chemiluminescence instrument.
如图3-4所示，准备20例交接药敏的样本，每例样本人工和自动化各铺3组药，每组药3复孔,药敏检测后统计出40例样本的Z值，Z值≥0.5即合格，计算得本发明一种类器官药敏检测自动化系统自动化稳定性与人工稳定性，可知本发明一种类器官药敏检测自动化系统各个自动作业子模块分工合作配合，不仅能够高效、快速的完成类器官从培养到药敏检测的一系列工作，有效避免了人工操作的高失误率以及不稳定性，且各个自动作业子模块分工互不影响，单个自动作业子模块故障不会影响另外自动作业子模块工作，在提高了工作效率的同时保证了稳定性，相较于效率低下的人工操作实验，能够进一步适用于大批量、大产量的批量类器官培养及药敏检测工作场合；另外，设备1人维护每月可处理300例样本，传统月产能300例则需要人工12人，节约费用为11人*10000元/月。As shown in Figure 3-4, prepare 20 samples for handing over drug susceptibility. Each sample is spread with 3 groups of drugs manually and automatically. Each group of drugs has 3 duplicate holes. After drug susceptibility testing, the Z value of 40 samples is calculated. Z If the value is ≥0.5, it is qualified. The automated stability and manual stability of the automated organoid drug sensitivity testing system of the present invention are calculated. It can be seen that the division of labor and cooperation of each automatic operation sub-module of the organoid drug sensitivity testing automated system of the present invention can not only be efficient, Quickly complete a series of tasks from organoid culture to drug sensitivity testing, effectively avoiding the high error rate and instability of manual operations, and the division of labor of each automatic operation sub-module does not affect each other, and the failure of a single automatic operation sub-module will not affect In addition, the automatic operation sub-module works to improve work efficiency while ensuring stability. Compared with inefficient manual experiments, it can be further suitable for large-scale, high-volume batch organoid culture and drug sensitivity testing workplaces; In addition, the equipment can be maintained by one person and can process 300 samples per month. The traditional monthly production capacity of 300 cases requires 12 workers, and the cost savings is 11 people * 10,000 yuan/month.
当然，以上所述仅是本发明的较佳实施方式，故凡依本发明专利申请范围所述的构造、特征及原理所做的等效变化或修饰，均包括于本发明专利申请范围内。Of course, the above are only preferred embodiments of the present invention. Therefore, any equivalent changes or modifications based on the structures, features and principles described in the patent application scope of the present invention are included in the patent application scope of the present invention.
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| CN117778166A (en) * | 2023-11-27 | 2024-03-29 | 创芯国际生物科技(广州)有限公司 | System and method for detecting drug sensitivity of organoid culture |
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