WO2023077598A1 - 一种自动化微生物涂布的装置和方法 - Google Patents

一种自动化微生物涂布的装置和方法 Download PDF

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Publication number
WO2023077598A1
WO2023077598A1 PCT/CN2021/135534 CN2021135534W WO2023077598A1 WO 2023077598 A1 WO2023077598 A1 WO 2023077598A1 CN 2021135534 W CN2021135534 W CN 2021135534W WO 2023077598 A1 WO2023077598 A1 WO 2023077598A1
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Prior art keywords
coating
hole
rod
sleeve
diameter
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English (en)
French (fr)
Inventor
张智彧
司同
蓝云泉
庞任维
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Shenzhen Institute of Advanced Technology of CAS
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Shenzhen Institute of Advanced Technology of CAS
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M33/00—Means for introduction, transport, positioning, extraction, harvesting, peeling or sampling of biological material in or from the apparatus
    • C12M33/02—Means for introduction, transport, positioning, extraction, harvesting, peeling or sampling of biological material in or from the apparatus by impregnation, e.g. using swabs or loops
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B08—CLEANING
    • B08B—CLEANING IN GENERAL; PREVENTION OF FOULING IN GENERAL
    • B08B1/00—Cleaning by methods involving the use of tools
    • B08B1/10—Cleaning by methods involving the use of tools characterised by the type of cleaning tool
    • B08B1/14—Wipes; Absorbent members, e.g. swabs or sponges
    • B08B1/143—Wipes
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B08—CLEANING
    • B08B—CLEANING IN GENERAL; PREVENTION OF FOULING IN GENERAL
    • B08B1/00—Cleaning by methods involving the use of tools
    • B08B1/30—Cleaning by methods involving the use of tools by movement of cleaning members over a surface
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B08—CLEANING
    • B08B—CLEANING IN GENERAL; PREVENTION OF FOULING IN GENERAL
    • B08B3/00—Cleaning by methods involving the use or presence of liquid or steam
    • B08B3/04—Cleaning involving contact with liquid
    • B08B3/08—Cleaning involving contact with liquid the liquid having chemical or dissolving effect
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M37/00—Means for sterilizing, maintaining sterile conditions or avoiding chemical or biological contamination
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M39/00—Means for cleaning the apparatus or avoiding unwanted deposits of microorganisms
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation

Definitions

  • the invention belongs to the technical field of microbial coating, in particular to an automatic microbial coating device and method.
  • the clone coating steps in the laboratory are mainly manually operated by experimental technicians; there are also some automatic cloning machines on the market for colony coating on square single-well plates, but this equipment is very expensive and can only complete 200 colonies per hour. For the coating of each sample, the coating efficiency also needs to be improved.
  • the purpose of the present invention is to provide a device and method for automatic microbial coating to solve the technical problems of low efficiency and high error rate of existing microbial coating technology.
  • the present invention provides a device for automatic microbial coating, comprising: a coating rod; the coating rod includes a coating rod and a sleeve that cooperate with each other;
  • the sleeve includes a sleeve body and a clamping part connected to each other; the clamping part is arranged on the upper end of the sleeve body; the diameter of the clamping part is larger than the diameter of the sleeve body; the sleeve is provided with a first through hole and a second through hole. Two through holes; the second through hole is located at the upper part of the sleeve, the first through hole is located at the lower part of the sleeve, and the diameter of the first through hole is smaller than the diameter of the second through hole;
  • the coating rod comprises a cylindrical coating rod main body, the lower end of the coating rod main body is equipped with a coating head, the upper end of the coating rod main body is provided with a guide rod, and the upper end of the guide rod passes through the first through hole and extends into the second In the through hole; the upper end of the guide rod is a threaded section; the threaded section is threaded with a nut, and the nut is located in the second through hole; the diameter of the nut is greater than the diameter of the first through hole, but smaller than the diameter of the second through hole.
  • a further improvement of the present invention lies in that: the length of the guide rod is greater than the length of the first through hole.
  • both the sleeve body and the clipping part are in the shape of a cylinder, and the clipping part is concentrically arranged on the upper end of the sleeve body; the diameter of the clipping part is larger than that of the sleeve body.
  • the further improvement of the present invention is: it also includes a carrier; the carrier includes a carrier body, and the carrier body includes a flat plate on the top and vertical plates supported on both sides of the flat plate; Three through holes; the diameter of the third through hole is larger than the diameter of the sleeve and smaller than the diameter of the gripping part.
  • the further improvement of the present invention is: it also includes a coating rod cleaning system;
  • the coating rod cleaning system includes a transmission mechanism and a brushing mechanism;
  • the transmission mechanism includes a fixed frame and a DC motor installed on the fixed frame; the rotating shaft of the DC motor protrudes from one side of the fixed frame and is fixedly connected with a turntable;
  • the scrubbing mechanism includes a brush, an alcohol box, a linear slide rail and a slider; the brush is set in the alcohol box, the bottom plate of the alcohol box is equipped with a slider, and the slider is installed on the corresponding linear slide rail; Rod link shaft; one end of the connecting rod is eccentrically hinged to the turntable, and the other end is hinged to the link shaft of the connecting rod.
  • the further improvement of the present invention is that: the turntable and the connecting rod form a crank linkage; the alcohol box can reciprocate along the direction of the linear slide rail under the drive of the turntable and the connecting rod.
  • the fixed frame includes two vertical fixed plates arranged in parallel; the top of the fixed plate is fixedly connected to the motor seat; the motor seat is flat; the lower surface of the motor seat is equipped with a stepping motor; the rotation of the stepping motor
  • the shaft extends out of the motor base and is fixedly connected with a baffle; a bending induction part is arranged on the baffle;
  • a first sensor and a second sensor are respectively fixed on the upper surface of the motor base on both sides of the rotating shaft; the first sensor and the second sensor are used to sense the position of the baffle by cooperating with the bending sensing part.
  • the further improvement of the present invention is that: several coating rods are placed on the carrier, and the carrier is fixed above the station of the brushing mechanism through the limit block; the baffle is located above the station of the brushing mechanism to press the coating rod; The coating head of the coating rod extends into the alcohol box; the coating head can be cleaned when the brush reciprocates.
  • the coating rod cleaning system further includes a sterilizing mechanism; the sterilizing mechanism is provided with a halogen lamp for drying and sterilizing the cleaned coating head.
  • the present invention provides a working method of a device for automatic microbial coating, comprising the following steps:
  • the robotic arm of the automated pipetting workstation draws the bacterial liquid from the source plate to the target plate;
  • the mechanical arm carries the coating rod and places it on the carrier; the mechanical arm clamps the carrier and moves to the brushing mechanism to clean the coating rod, and then the mechanical arm clamps the carrier to the sterilization mechanism for coating Sticks were dried and sterilized.
  • the invention establishes an automatic technological process from liquid sample operation to microorganism coating by means of the developed, designed and customized coating device.
  • the 8-channel pipetting mechanical arm still loads the coating rod to coat the corresponding orifice plate.
  • the coating After the coating is completed, it returns to the cleaning mechanism in the coating device to clean the coating rod. After cleaning, it enters the sterilization mechanism. High temperature drying, sterilization, and then prepare for the next experiment.
  • the whole coating process has a high degree of automation, which reduces the errors caused by human factors.
  • the invention provides a device for automatic microbial coating, which includes a new coating rod structure;
  • the coating rod includes a coating rod and a sleeve that cooperate with each other;
  • the sleeve includes a sleeve main body and a clamping part that are connected to each other; the clamping The part is arranged on the upper end of the sleeve main body; the diameter of the clamping part is larger than the diameter of the sleeve main body; the first through hole and the second through hole are provided in the sleeve; the second through hole is located on the upper part of the sleeve, and the first through hole The hole is located at the lower part of the sleeve, and the diameter of the first through hole is smaller than the diameter of the second through hole;
  • the coating rod comprises a cylindrical coating rod main body, and the lower end of the coating rod main body is equipped with a coating head, and the coating rod main body
  • the upper end of the guide rod is provided with a guide rod, and
  • the present invention can determine the optimal cleaning and sterilization time through experiments, which can meet the above-mentioned automated microbial coating, high throughput, convenient operation, wide application range, and low overall cost. It can be seen that the present invention The process involved can completely replace the traditional manual coating, which promotes the construction of a high-throughput automation platform.
  • the present invention designs and customizes the coating device based on the automatic liquid transfer workstation.
  • a certain amount of bacterial liquid is drawn by the automatic liquid transfer workstation, and the liquid is transferred to the agar plate to be coated, and then the microorganism is coated by the coating device.
  • the sample is evenly dispersed on the surface of the medium plate, and finally the coating rod is cleaned and sterilized by the cleaning device to realize repeated use and prevent cross-contamination.
  • the device improves the speed and precision of the microbial plate coating operation, and liberates technicians from a large amount of repetitive work.
  • the throughput of monoclonal coating is 1 hour to complete the processing of not less than 300 samples.
  • the throughput of monoclonal coating in the present invention is increased by more than 1.5 times compared with existing commercialized instruments.
  • the device of the present invention can be integrated with any major brand of automatic liquid workstation, has a wide range of applications, and does not affect the original functions of the automatic liquid workstation.
  • Fig. 1 is the structural representation of coating bar among the present invention
  • Fig. 2 is the structural representation of coating rod and carrier of the present invention
  • Fig. 3 is a schematic structural view of a device for automatic microbial coating of the present invention.
  • Fig. 4 is the structural representation of transmission mechanism
  • Fig. 5 is the structural representation of scrubbing mechanism
  • FIG. 6 is a bottom view of the scrubbing mechanism shown in FIG. 5 .
  • this embodiment discloses an automatic microorganism coating device, including: several coating rods 20 and a coating rod cleaning system.
  • the coating rod 20 includes a coating rod 201 and a sleeve 202 that cooperate with each other.
  • the sleeve 202 includes a sleeve main body 2021 and a clamping portion 2022 connected to each other; the sleeve main body 2021 is cylindrical, and the clamping portion 2022 is also cylindrical, and is concentrically arranged on the upper end of the sleeve main body 2021; the clamping portion 2022 The diameter is greater than the diameter of the sleeve body 2021 .
  • the sleeve 202 is provided with a through first through hole 2024 and a second through hole 2023; the second through hole 2023 is located at the upper part of the sleeve 202, the first through hole 2024 is located at the lower part of the sleeve 202, and the diameter of the first through hole 2024 smaller than the diameter of the second through hole 2023 .
  • Coating bar 201 comprises cylindrical coating bar main body 2010, and coating head 2011 is installed on the lower end of coating bar main body 2010, and the upper end of coating bar main body 2010 is provided with guide bar 2012, and the upper end of guide bar 2012 passes through the first A through hole 2024 extends into the second through hole 2023; the upper end of the guide rod 2012 is a threaded section 2013; The diameter of the nut 2014 is larger than the diameter of the first through hole 2024 and smaller than the diameter of the second through hole 2023 .
  • the guide rod 2012 is in clearance fit with the first through hole 2024 .
  • the shape of the coating head 2011 shown in FIG. 1 is spherical; its shape can also be selected according to the specific culture medium and culture plate, such as rod-shaped, flat-bottomed and other shapes.
  • FIG. 2 Please refer to Fig. 2, several coating rods 20 can be placed on the carrier 21; ;
  • the plate 2100 is provided with a number of third through holes 2102; A plurality of coating rods 20 can be inserted into the corresponding third through holes 2102, so that the clamping portion 2022 is exposed on the upper part of the plate 2100 for the mechanical arm of the automatic liquid pipetting workstation to clamp.
  • the bottom of the vertical plate 2101 is provided with a large bottom plate 23 .
  • a number of limit blocks 13 are provided on the workbench of the automated liquid pipetting workstation; the large bottom plate 23 at the bottom of the vertical plate 2101 can be stuck in the limit blocks 13 to keep the carrier 21 at a certain position.
  • the coating rod cleaning system includes a transmission mechanism 101 , a scrubbing mechanism 102 and a sterilization mechanism 103 .
  • the transmission mechanism 101 , the scrubbing mechanism 102 and the sterilizing mechanism 103 are arranged on the working plate 100 of the automatic pipetting workstation.
  • the transmission mechanism 101 includes a fixed frame, a baffle 1 , a stepping motor 6 and a DC motor 7 .
  • the fixing frame includes two vertical fixing plates 1011 arranged in parallel; the two vertical fixing plates 1011 arranged in parallel are fixed on the working plate 100 in parallel, and the top is fixedly connected to the motor base 5 .
  • the motor base 5 is flat.
  • a stepping motor 6 is installed on the lower surface of the motor base 5; the rotating shaft 2 of the stepping motor 6 protrudes from the motor base 5 and is fixedly connected with the baffle 1; the baffle 1 is provided with a bending induction part 1012.
  • a first sensor 3 and a second sensor 4 are respectively fixed on the upper surface of the motor base 5 on both sides of the rotating shaft 2 .
  • the rotation of the stepping motor 6 can drive the baffle 1 to rotate through the rotating shaft 2 , and the position of the baffle 1 can be sensed through the first sensor 3 , the second sensor 4 and the bending sensing part 1012 .
  • the DC motor 7 is installed between two vertical fixing plates 1011, and the rotating shaft of the DC motor 7 extends out of one side of the vertical fixing plate 1011, and is fixedly connected with the rotating disk 8; the connecting rod 9 is eccentrically installed on the rotating disk 8.
  • the rotation of the DC motor 7 can drive the connecting rod 9 to reciprocate.
  • the fixed frame is covered with a motor cover 22 .
  • the scrubbing mechanism 102 includes a brush 10 , an alcohol box 11 , a linear slide rail 16 and a slider 17 .
  • the brush 10 is arranged in the alcohol box 11, and the bottom plate 12 of the alcohol box 11 is equipped with two sliders 17 in parallel, and the two sliders 17 are installed on two linear slide rails 16; the linear slide rails 16 are fixed on the working plate 100 superior.
  • One side of the base plate 12 is equipped with a connecting rod link shaft 14 .
  • One end of the connecting rod 9 is hinged with the rotating disk 8, and the other end is hinged with the connecting rod link shaft 14.
  • coating rods 20 are placed on the carrier 21, and the carrier 21 is fixed above the station of the brushing mechanism 102 through the limit block 13; when the DC motor 7 rotates, it drives the turntable 8 to rotate, and then the alcohol box can be driven by the connecting rod 9 11 Under the cooperation of the slide block 17 and the linear slide rail 16, reciprocating linear motion is performed, and then the coating head 2011 of the coating rod 20 is cleaned by the brush 10.
  • the stepper motor 6 in the transmission mechanism 101 rotates to drive the baffle 1 to rotate through the rotating shaft 2, and cooperates with the bending sensing part 1012 through the first sensor 3 and the second sensor 4 to sense the position of the baffle 1, so that When baffle plate 1 is positioned at the station top of scrubbing mechanism 102 and presses carrier 21, stops rotating; Effect.
  • the station below of brushing mechanism 102 is also equipped with sensor seat 18, and the 3rd sensor 15 is installed in sensor 18;
  • the bottom edge of alcohol box 11 is equipped with trigger piece 19; The working limit position of induction alcohol box 11.
  • a halogen lamp 24 is provided in the sterilization mechanism 103 .
  • Halogen lamp 24 is fixed on the work plate 100, can heat the coated rod 20 that is positioned at the sterilizing mechanism 103 tops cleaned; Drying temperature is more than 500 degrees Celsius in the sterilizing mechanism 103, to dry the coated rod 20, Sterilization: the dried and sterilized coating rod 20 can be grasped by the mechanical arm as a whole and moved to the working position for standby.
  • This embodiment discloses a working method of an automatic microorganism coating device, comprising the following steps:
  • the 8-channel robotic arm of the automated pipetting workstation draws a certain amount of bacterial liquid from the source plate to the target plate;
  • the 8-channel pipetting mechanical arm knocks off the suction head, it moves to the carrier 21 to load a clean coating rod 20, and then moves to the target plate to perform a moving action to complete the coating; the coating in the coating rod 20 Due to the effect of gravity, the rod 201 is generally at the lowest working position; some culture medium fluctuates up and down, and the coating rod 201 will be squeezed upward and stretched according to the height fluctuation of the culture medium, without destroying the bacterial liquid itself, so as to ensure the accuracy of the experiment;
  • the flexible profiling design of the coating rod 20 to different heights of the medium, the coating rod and the sleeve can move relatively, can adapt to the unevenness of the solid medium in the orifice plate, and move up and down relative to the height of the medium, so that The bacteria liquid is very evenly distributed in the orifice plate;
  • the 8-channel pipetting robot arm carries the coating rod and moves to the scrubbing mechanism 102 for cleaning, and then goes to the sterilization mechanism 103 for drying and sterilizing at high temperature, and finally the 8-channel liquid pipetting robot arm moves to the carrier for cleaning.
  • the applicator bar is unloaded and the applicator bar is left to dry.

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Abstract

一种自动化微生物涂布的装置和方法；所述装置，包括：涂布棒；涂布棒包括相互配合的涂布杆和套筒；套筒包括相互连接的套筒主体和夹取部；夹取部设置于套筒主体的上端；夹取部的直径大于套筒主体的直径；套筒中设有贯通的第一通孔和第二通孔；第一通孔的直径小于第二通孔的直径；涂布杆包括圆柱体状的涂布杆主体，涂布杆主体的下端安装有涂布头，涂布杆主体的上端设有导向杆，导向杆的上端穿过第一通孔伸入第二通孔中；导向杆的上端为螺纹段；螺纹段上螺纹连接有螺母，所述螺母位于第二通孔中。

Description

一种自动化微生物涂布的装置和方法 技术领域
本发明属于微生物涂布技术领域，特别涉及一种自动化微生物涂布的装置和方法。
 背景技术
将微生物样品通过固体琼脂培养基铺板操作获得单克隆菌落，是微生物研究中的关键工艺。传统的自动化涂板需经过移液、涂布等步骤实现，目前主要由技术人员手持涂布棒来回移动进行涂布，标准化水平低；若需要涂布大量的样品，则非常费时，且技术人员易疲劳，往往会出错。
目前实验室的克隆涂布步骤主要是实验技术人员手动操作；市场上也出现了一些自动化挑选克隆仪对方形单孔板进行菌落涂布，但是这种设备特别昂贵，而且1小时只能完成200个样品的涂布，涂布效率也待提升。
由于微生物涂布在相关分子生物学实验流程中往往成为实验通量的瓶颈所在，因此迫切需要一种装置和工艺实现微生物自动化涂布，大幅提高实验通量和标准化水平，降低错误率。
 技术问题
本发明的目的在于提供一种动化微生物涂布的装置和方法，以解决现有微生物涂布技术效率低、错误率高的技术问题。
 技术解决方案
为了实现上述目的，本发明采用如下技术方案：
第一方面，本发明提供一种自动化微生物涂布的装置，包括：涂布棒；涂布棒包括相互配合的涂布杆和套筒；
套筒包括相互连接的套筒主体和夹取部；夹取部设置于套筒主体的上端；夹取部的直径大于套筒主体的直径；套筒中设有贯通的第一通孔和第二通孔；第二通孔位于套筒上部，第一通孔位于套筒下部，且第一通孔的直径小于第二通孔的直径；
涂布杆包括圆柱体状的涂布杆主体，涂布杆主体的下端安装有涂布头，涂布杆主体的上端设有导向杆，导向杆的上端穿过第一通孔伸入第二通孔中；导向杆的上端为螺纹段；螺纹段上螺纹连接有螺母，所述螺母位于第二通孔中；螺母的直径大于第一通孔的直径，而小于第二通孔的直径。
本发明进一步的改进在于：导向杆的长度大于第一通孔的长度。
本发明进一步的改进在于：套筒主体和夹取部均呈圆柱体状，夹取部同心设置于套筒主体的上端；夹取部的直径大于套筒主体的直径。
本发明进一步的改进在于：还包括载架；载架包括载架主体，载架主体包括位于顶部的平板以及支撑在平板两侧的竖板；平板上设有若干用于放置涂布棒的第三通孔；第三通孔的直径大于套筒的直径，小于夹取部的直径。
本发明进一步的改进在于：还包括涂布棒清洁系统；所述涂布棒清洁系统包括传动机构和刷洗机构；
传动机构包括固定框架和安装于固定框架上的直流电机；直流电机的转轴伸出固定框架一侧固定连接有转盘；
刷洗机构包括毛刷、酒精盒、直线滑轨和滑块；毛刷设置于酒精盒中，酒精盒的底板安装有滑块，滑块安装在对应直线滑轨上；底板的一侧安装有连杆链接轴；连杆一端与转盘偏心铰接，另一端与连杆链接轴铰接。
本发明进一步的改进在于：转盘和连杆构成曲柄连杆机构；酒精盒能够在转盘和连杆的驱动下沿直线滑轨方向进行往复运动。
本发明进一步的改进在于：固定框架包括两块平行设置的竖向固定板；固定板顶部固定连接电机座；电机座为平板状；电机座的下表面安装有步进电机；步进电机的旋转轴伸出电机座固定连接有挡板；挡板上设有一个弯折感应部；
电机座上表面位于旋转轴两侧分别固定有第一传感器和第二传感器；所述第一传感器和第二传感器用于通过与弯折感应部相配合，感应挡板的位置。
本发明进一步的改进在于：若干涂布棒放置于载架上，载架通过限位块固定在刷洗机构的工位上方；所述挡板位于刷洗机构的工位上方压住涂布棒；若干涂布棒的涂布头伸入所述酒精盒中；所述毛刷往复运动时能够清洁所述涂布头。
本发明进一步的改进在于：所述涂布棒清洁系统还包括灭菌机构；所述灭菌机构中设有用于对清洁过的涂布头进行干燥、灭菌的卤素灯。
第二方面，本发明提供一种自动化微生物涂布的装置的工作方法，包括以下步骤：
S1、自动化移液工作站的机械臂从来源板中吸取菌液到目标板中；
S2、机械臂打掉吸头后，移动到载架上加载若干洁净的涂布棒，然后移动到目标板上进行移动动作来完成涂布；
S3、完成涂布后，机械臂携带涂布棒放置到载架上；机械臂夹持载架移动到刷洗机构对涂布棒进行清洗，然后机械臂夹持载架到灭菌机构对涂布棒进行干燥、灭菌。
本发明借助开发并设计、定制的涂布装置，建立了一个从液体样品操作到微生物涂布自动化的工艺过程。首先由工程技术人员编写软件驱动和工艺脚本，然后由自动化移液工作站的8通道移液机械臂加载吸头吸取一定量的菌液，放液至方孔板或者6孔板或者其它培养板，然后仍然由8通道移液机械臂加载涂布棒进行相应孔板的涂布，涂布完成后回到涂布装置内的清洗机构内，进行涂布棒的清洗，清洗完成后进入灭菌机构高温干燥、灭菌，进而为下次的实验做准备。整个涂布过程自动化程度高，减少了人为因素带来的误差。
 有益效果
本发明提供一种自动化微生物涂布的装置，设置新的涂布棒结构；涂布棒包括相互配合的涂布杆和套筒；套筒包括相互连接的套筒主体和夹取部；夹取部设置于套筒主体的上端；夹取部的直径大于套筒主体的直径；套筒中设有贯通的第一通孔和第二通孔；第二通孔位于套筒上部，第一通孔位于套筒下部，且第一通孔的直径小于第二通孔的直径；涂布杆包括圆柱体状的涂布杆主体，涂布杆主体的下端安装有涂布头，涂布杆主体的上端设有导向杆，导向杆的上端穿过第一通孔伸入第二通孔中；导向杆的上端为螺纹段；螺纹段上螺纹连接有螺母，所述螺母位于第二通孔中；螺母的直径大于第一通孔的直径，而小于第二通孔的直径；涂布棒可以通过机械臂批量夹取，批量进行菌液的涂布；相对于现有技术，本发明能够很好替代传统的手动涂布，解决了现有技术存在的单克隆数量少和微生物涂布过程耗时的问题，其整体工艺流程简单，实际操作方便，适用性广。
进一步的，本发明能够通过实验确定最佳清洗和灭菌时间，能够满足对于上述中自动化的微生物涂布，通量高，操作方便，且适用范围广，整体成本低，由此可见，本发明涉及的工艺完全可以代替传统的手动涂布，促进了高通量自动化平台的建设。
进一步的，本发明基于自动化移液工作站设计、定制了涂布装置，首先由自动化移液工作站吸取一定量的菌液，并移液至需要涂布的琼脂平板上，然后利用涂布装置将微生物样品均匀分散在培养基平板表面，最后使用清洗装置对涂布棒进行清洗灭菌实现反复利用，防止交叉污染。本装置提高了微生物平板涂布操作的速度和精度，将技术人员从大量重复性工作中解放出来。本发明中单克隆涂布通量为1小时可完成不少于300个样品的处理。本发明中单克隆涂布通量与现有商业化仪器相比提高了1.5倍以上。同时，本发明装置可以与任何主要品牌的自动化液体工作站整合使用，应用范围广，且不影响自动化液体工作站的原有功能。
 附图说明
构成本发明的一部分的说明书附图用来提供对本发明的进一步理解，本发明的示意性实施例及其说明用于解释本发明，并不构成对本发明的不当限定。在附图中：
图1为本发明中涂布棒的结构示意图；
图2为本发明涂布棒和载架的结构示意图；
图3为本发明一种自动化微生物涂布的装置的结构示意图；
图4为传动机构的结构示意图；
图5为刷洗机构的结构示意图；
图6为图5所示刷洗机构的底部视图。
 本发明的最佳实施方式
下面将参考附图并结合实施例来详细说明本发明。需要说明的是，在不冲突的情况下，本发明中的实施例及实施例中的特征可以相互组合。
以下详细说明均是示例性的说明，旨在对本发明提供进一步的详细说明。除非另有指明，本发明所采用的所有技术术语与本发明所属领域的一般技术人员的通常理解的含义相同。本发明所使用的术语仅是为了描述具体实施方式，而并非意图限制根据本发明的示例性实施方式。
实施例1
请参阅图1至图6所示，本实施例公开一种自动化微生物涂布的装置，包括：若干涂布棒20以及涂布棒清洁系统。
请参阅图1所示，涂布棒20包括相互配合的涂布杆201和套筒202。套筒202包括相互连接的套筒主体2021和夹取部2022；套筒主体2021呈圆柱体状，夹取部2022也呈圆柱体状，同心设置于套筒主体2021的上端；夹取部2022的直径大于套筒主体2021的直径。套筒202中设有贯通的第一通孔2024和第二通孔2023；第二通孔2023位于套筒202上部，第一通孔2024位于套筒202下部，且第一通孔2024的直径小于第二通孔2023的直径。
涂布杆201包括圆柱体状的涂布杆主体2010，涂布杆主体2010的下端安装有涂布头2011，涂布杆主体2010的上端设有导向杆2012，导向杆2012的上端穿过第一通孔2024伸入第二通孔2023中；导向杆2012的上端为螺纹段2013；螺纹段2013上螺纹连接有螺母2014，所述螺母位于第二通孔2023中。螺母2014的直径大于第一通孔2024的直径，而小于第二通孔2023的直径。导向杆2012与第一通孔2024间隙配合。图1所示涂布头2011的形状为球状；其形状也可以根据具体的培养基和培养板进行选择，例如棒状、平底装等形状。
请参阅图2所示，若干涂布棒20可以放置于载架21上；载架21包括载架主体210，载架主体210包括位于顶部的平板2100以及支撑在平板2100两侧的竖板2101；平板2100上设有若干第三通孔2102；第三通孔2102的直径大于套筒202的直径，小于夹取部2022的直径。若干涂布棒20可以插于对应的第三通孔2102中，而使夹取部2022露在平板2100上部，以供自动化移液工作站的机械臂夹取。竖板2101的底部设有大底板23。自动化移液工作站的工作台上设有若干限位块13；竖板2101底部的大底板23能够卡在限位块13中，而将载架21保持在某一工位上。
请参阅图3所示，涂布棒清洁系统包括传动机构101、刷洗机构102和灭菌机构103。传动机构101、刷洗机构102和灭菌机构103设置在自动化移液工作站的工作板100上。
请参阅图4所示，传动机构101包括固定框架、挡板1、步进电机6和直流电机7。固定框架包括两块平行设置的竖向固定板1011；两块平行设置的竖向固定板1011平行固定在工作板100上，顶部固定连接电机座5。电机座5为平板状。电机座5的下表面安装有步进电机6；步进电机6的旋转轴2伸出电机座5与挡板1固定连接；挡板1上设有一个弯折感应部1012。电机座5上表面位于旋转轴2两侧分别固定有第一传感器3和第二传感器4。步进电机6旋转能够通过旋转轴2带动挡板1旋转，通过第一传感器3和第二传感器4与弯折感应部1012能够感应出挡板1的位置。直流电机7安装在于两块竖向固定板1011之间，直流电机7的转轴伸出一侧竖向固定板1011，并固定连接有转盘8；转盘8上偏心安装有连杆9。直流电机7的转动能够驱动连杆9往复运动。固定框架外罩有电机罩22。
请参阅图5和图6所示，刷洗机构102包括毛刷10、酒精盒11、直线滑轨16和滑块17。毛刷10设置于酒精盒11中，酒精盒11的底板12下部平行安装有两个滑块17，两个滑块17安装在两个直线滑轨16上；直线滑轨16固定在工作板100上。底板12的一侧安装有连杆链接轴14。连杆9一端与转盘8铰接，另一端与连杆链接轴14铰接。若干涂布棒20放置于载架21上，载架21通过限位块13固定在刷洗机构102的工位上方；直流电机7转动时，带动转盘8旋转，进而通过连杆9能够驱动酒精盒11在滑块17、直线滑轨16的配合下，进行往复直线运动，进而通过毛刷10清洗涂布棒20的涂布头2011。清洗前，传动机构101中的步进电机6旋转通过旋转轴2带动挡板1旋转，通过第一传感器3和第二传感器4与弯折感应部1012配合，感应出挡板1的位置，使挡板1位于刷洗机构102的工位上方压住载架21时，停止转动；这样毛刷10清洗涂布棒20的涂布头2011时，涂布棒20的位置能够保持稳定，保证清洗的效果。刷洗机构102的工位下方还安装有传感器座18，传感器做18中安装有第三传感器15；酒精盒11的底部边缘安装有触发片19；通过触发片19和第三传感器15相互配合，能够感应酒精盒11的工作极限位置。
灭菌机构103中设有卤素灯24。卤素灯24固定在工作板100上，能够对位于灭菌机构103上方的清洗过的涂布棒20进行加热；灭菌机构103中干燥温度为500摄氏度以上，以对涂布棒20进行干燥、灭菌；干燥、灭菌后的涂布棒20可以通过机械臂整体抓取移到待工作位置进行备用。
实施例2
本实施例公开一种自动化微生物涂布的装置的工作方法，包括以下步骤：
S1、自动化移液工作站的8通道机械臂从来源板中吸取一定量的菌液到目标板中；
S2、8通道移液机械臂打掉吸头后，移动到载架21上加载洁净的涂布棒20，然后移动到目标板上进行移动动作来完成涂布；涂布棒20中的涂布杆201由于重力作用，一般都在最低的工作位；有些培养基高低起伏，涂布杆201会根据培养基的高低起伏被挤压向上伸缩，不会破坏菌液本身，保证实验的准确性；涂布棒20对不同高低培养基的柔性仿形的设计，涂布杆和套筒能够相对运动，能够适应孔板中固体培养基不平的情况，根据培养基的高低进而相对上下运动，从而使菌液非常均匀的分布在孔板内；
S3、完成涂布后，8通道移液机械臂携带涂布棒移动到刷洗机构102进行清洗，然后到灭菌机构103高温进行干燥、灭菌，最后8通道移液机械臂移动到载架将涂布棒卸载掉，然后涂布棒进行静置晾干。
由技术常识可知，本发明可以通过其它的不脱离其精神实质或必要特征的实施方案来实现。因此，上述公开的实施方案，就各方面而言，都只是举例说明，并不是仅有的。所有在本发明范围内或在等同于本发明的范围内的改变均被本发明包含。

Claims (10)

  1. 一种自动化微生物涂布的装置，其特征在于，包括：涂布棒（20）；涂布棒（20）包括相互配合的涂布杆（201）和套筒（202）；
    套筒（202）包括相互连接的套筒主体（2021）和夹取部（2022）；夹取部设置于套筒主体（2021）的上端；夹取部（2022）的直径大于套筒主体（2021）的直径；套筒（202）中设有贯通的第一通孔（2024）和第二通孔（2023）；第二通孔（2023）位于套筒（202）上部，第一通孔（2024）位于套筒（202）下部，且第一通孔（2024）的直径小于第二通孔（2023）的直径；
    涂布杆（201）包括圆柱体状的涂布杆主体（2010），涂布杆主体（2010）的下端安装有涂布头（2011），涂布杆主体（2010）的上端设有导向杆（2012），导向杆（2012）的上端穿过第一通孔（2024）伸入第二通孔（2023）中；导向杆（2012）的上端为螺纹段（2013）；螺纹段（2013）上螺纹连接有螺母（2014），所述螺母位于第二通孔（2023）中；螺母（2014）的直径大于第一通孔（2024）的直径，而小于第二通孔（2023）的直径。
  2. 根据权利要求1所述的一种自动化微生物涂布的装置，其特征在于，导向杆（2012）的长度大于第一通孔（2024）的长度。
  3. 根据权利要求1所述的一种自动化微生物涂布的装置，其特征在于，套筒主体（2021）和夹取部（2022）均呈圆柱体状，夹取部（2022）同心设置于套筒主体（2021）的上端；夹取部（2022）的直径大于套筒主体（2021）的直径。
  4. 根据权利要求1所述的一种自动化微生物涂布的装置，其特征在于，还包括载架（21）；载架（21）包括载架主体（210），载架主体（210）包括位于顶部的平板（2100）以及支撑在平板（2100）两侧的竖板（2101）；平板（2100）上设有若干用于放置涂布棒（20）的第三通孔（2102）；第三通孔（2102）的直径大于套筒（202）的直径，小于夹取部（2022）的直径。
  5. 根据权利要求1所述的一种自动化微生物涂布的装置，其特征在于，还包括涂布棒清洁系统；所述涂布棒清洁系统包括传动机构（101）和刷洗机构（102）；
    传动机构（101）包括固定框架和安装于固定框架上的直流电机（7）；直流电机（7）的转轴伸出固定框架一侧固定连接有转盘（8）；
    刷洗机构（102）包括毛刷（10）、酒精盒（11）、直线滑轨（16）和滑块（17）；毛刷（10）设置于酒精盒（11）中，酒精盒（11）的底板（12）安装有滑块（17），滑块（17）安装在对应直线滑轨（16）上；底板（12）的一侧安装有连杆链接轴（14）；连杆（9）一端与转盘（8）偏心铰接，另一端与连杆链接轴（14）铰接。
  6. 根据权利要求5所述的一种自动化微生物涂布的装置，其特征在于，转盘（8）和连杆（9）构成曲柄连杆机构；酒精盒（11）能够在转盘（8）和连杆（9）的驱动下沿直线滑轨（16）方向进行往复运动。
  7. 根据权利要求5所述的一种自动化微生物涂布的装置，其特征在于，固定框架包括两块平行设置的竖向固定板（1011）；固定板（1011）顶部固定连接电机座（5）；电机座（5）为平板状；电机座（5）的下表面安装有步进电机（6）；步进电机（6）的旋转轴（2）伸出电机座（5）固定连接有挡板（1）；挡板（1）上设有一个弯折感应部（1012）；
    电机座（5）上表面位于旋转轴（2）两侧分别固定有第一传感器（3）和第二传感器（4）；所述第一传感器（3）和第二传感器（4）用于通过与弯折感应部（1012）相配合，感应挡板（1）的位置。
  8. 根据权利要求7所述的一种自动化微生物涂布的装置，其特征在于，若干涂布棒（20）放置于载架（21）上，载架（21）通过限位块（13）固定在刷洗机构（102）的工位上方；所述挡板（1）位于刷洗机构（102）的工位上方压住涂布棒（20）；若干涂布棒（20）的涂布头（2011）伸入所述酒精盒（11）中；所述毛刷（10）往复运动时能够清洁所述涂布头（2011）。
  9. 根据权利要求5所述的一种自动化微生物涂布的装置，其特征在于，所述涂布棒清洁系统还包括灭菌机构（103）；所述灭菌机构（103）中设有用于对清洁过的涂布头（2011）进行干燥、灭菌的卤素灯（24）。
  10. 一种自动化微生物涂布的装置的工作方法，其特征在于，基于权利要求9所述的一种自动化微生物涂布的装置，包括以下步骤：
    S1、自动化移液工作站的机械臂从来源板中吸取菌液到目标板中；
    S2、机械臂打掉吸头后，移动到载架上加载若干洁净的涂布棒（20），然后移动到目标板上进行移动动作来完成涂布；
    S3、完成涂布后，机械臂携带涂布棒放置到载架上；机械臂夹持载架移动到刷洗机构（102）对涂布棒（20）进行清洗，然后机械臂夹持载架到灭菌机构（103）对涂布棒（20）进行干燥、灭菌。
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