WO2023088014A1 - 转移式高通量超薄切片染色仪 - Google Patents

转移式高通量超薄切片染色仪 Download PDF

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WO2023088014A1
WO2023088014A1 PCT/CN2022/125798 CN2022125798W WO2023088014A1 WO 2023088014 A1 WO2023088014 A1 WO 2023088014A1 CN 2022125798 W CN2022125798 W CN 2022125798W WO 2023088014 A1 WO2023088014 A1 WO 2023088014A1
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rinsing
bottle
staining
manipulator
type high
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PCT/CN2022/125798
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English (en)
French (fr)
Inventor
梁静南
色日格楞
罗元明
李春立
樊峥
赵彤
孙树涛
王前
张晓兰
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中国科学院微生物研究所
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Publication of WO2023088014A1 publication Critical patent/WO2023088014A1/zh

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    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N1/00—Sampling; Preparing specimens for investigation
    • G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
    • G01N1/30—Staining; Impregnating ; Fixation; Dehydration; Multistep processes for preparing samples of tissue, cell or nucleic acid material and the like for analysis
    • G01N1/31—Apparatus therefor
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N23/00—Investigating or analysing materials by the use of wave or particle radiation, e.g. X-rays or neutrons, not covered by groups G01N3/00 – G01N17/00, G01N21/00 or G01N22/00
    • G01N23/02—Investigating or analysing materials by the use of wave or particle radiation, e.g. X-rays or neutrons, not covered by groups G01N3/00 – G01N17/00, G01N21/00 or G01N22/00 by transmitting the radiation through the material
    • G01N23/04—Investigating or analysing materials by the use of wave or particle radiation, e.g. X-rays or neutrons, not covered by groups G01N3/00 – G01N17/00, G01N21/00 or G01N22/00 by transmitting the radiation through the material and forming images of the material
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N23/00—Investigating or analysing materials by the use of wave or particle radiation, e.g. X-rays or neutrons, not covered by groups G01N3/00 – G01N17/00, G01N21/00 or G01N22/00
    • G01N23/20—Investigating or analysing materials by the use of wave or particle radiation, e.g. X-rays or neutrons, not covered by groups G01N3/00 – G01N17/00, G01N21/00 or G01N22/00 by using diffraction of the radiation by the materials, e.g. for investigating crystal structure; by using scattering of the radiation by the materials, e.g. for investigating non-crystalline materials; by using reflection of the radiation by the materials
    • G01N23/20008—Constructional details of analysers, e.g. characterised by X-ray source, detector or optical system; Accessories therefor; Preparing specimens therefor
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N23/00—Investigating or analysing materials by the use of wave or particle radiation, e.g. X-rays or neutrons, not covered by groups G01N3/00 – G01N17/00, G01N21/00 or G01N22/00
    • G01N23/20—Investigating or analysing materials by the use of wave or particle radiation, e.g. X-rays or neutrons, not covered by groups G01N3/00 – G01N17/00, G01N21/00 or G01N22/00 by using diffraction of the radiation by the materials, e.g. for investigating crystal structure; by using scattering of the radiation by the materials, e.g. for investigating non-crystalline materials; by using reflection of the radiation by the materials
    • G01N23/20058—Measuring diffraction of electrons, e.g. low energy electron diffraction [LEED] method or reflection high energy electron diffraction [RHEED] method
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N35/00—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor
    • G01N35/0099—Automatic analysis not limited to methods or materials provided for in any single one of groups G01N1/00 - G01N33/00; Handling materials therefor comprising robots or similar manipulators

Definitions

  • the invention belongs to the technical field of transmission electron microscope.
  • ultra-thin section sample preparation technology When using transmission electron microscope to observe the inner ultrastructure of biological sample cells, it is necessary to use ultra-thin section sample preparation technology to make the sample into ultra-thin slices with a thickness of less than 100nm, and carry them on a tube with a diameter of about 3mm, an inner diameter of about 2.6nm, and a ring width of About 0.2mm and 0.02mm thickness of the grid, and then use tweezers to clamp the ring width position of the grid as much as possible (to avoid mechanical damage to the biological section to the greatest extent), and perform the staining operation before microscopic imaging observe.
  • This dyeing process requires the dyeing of two dye solutions before and after. After each dye solution is dyed, it needs to be rinsed and air-dried, a total of 6 steps.
  • the dyeing instrument uses two symmetrical clamping batch grid-carrying racks as the core functional components of the invention patent ZL 201610366342.4 and the utility model ZL 202020027034.0. On this basis, a mechanical transfer working mode is established to complete the loading The batch-loaded nets inserted on the grid racks are automatically operated for dyeing, rinsing, air-drying, re-dyeing, rinsing, and air-drying.
  • the invention provides a transfer type high-throughput ultra-thin section dyeing instrument, comprising: a support platform; a support column, the support column is at the center of the support platform and fixed on the support platform; a bottle cap platform, the bottle cap platform The central position of the center is fixed on the other end of the support column; the rinsing pool, staining bottle A and staining bottle B, the rinsing pool, staining bottle A and staining bottle B are all fixed on the support platform and around the support column.
  • the bottle cap platform there are three stations on the bottle cap platform, wherein the first station is a hollow hole position, and the second station and the third station are connected with the bottle caps of dye liquor bottles A and B. Size and dimensions match.
  • the rinsing tank includes: a rinsing liquid chamber A, a rinsing liquid inlet pipe interface, a connecting wall, a rinsing liquid chamber B, and a waste liquid outlet; one of the rinsing liquid chambers A There is a rinsing liquid inlet pipe interface on one side, and the other side is connected to the rinsing liquid chamber B through a connecting wall; the bottom of the rinsing liquid chamber B has a waste liquid outlet; the connecting wall has a number of small holes or slits, The liquid introduced from the rinsing liquid chamber A enters the rinsing liquid chamber B through the connecting wall in a direction close to parallel, so that the rinsing liquid can be rinsed in a direction nearly perpendicular to the grid carrier; the waste liquid outlet is connected to the waste liquid pool Same and realize the waste liquid flows into the waste liquid pool.
  • a waste liquid pool is further included, the waste liquid pool is under the supporting platform, and is used for receiving the liquid flowing out from the waste liquid outlet of the rinsing tank.
  • the present invention also includes: a manipulator and 3 stepper motors; the manipulator and 3 stepper motors are arranged above the support platform, and 3 stepper motors control the control manipulator in X, Y, Z Movement in three directions, and then realize the gripping and movement of the article by the manipulator; the manipulator is used to grab and place the bottle caps of the dye solution bottle A and the dye solution bottle B, and realize the operation of capping and uncapping; The manipulator can also grab and place the top cover of the functional parts of the loaded grid carrier; the stepper motor is arranged on the keel frame.
  • the air-drying system includes four-corner and top fans.
  • the present invention also provides a dyeing method for a transfer-type high-throughput ultra-thin section dyeing instrument, which includes the following steps: the manipulator grabs the bottle cap A' on the dye solution bottle A and leaves the dye solution bottle; the manipulator grabs the functional part S and enters In the dye solution A; after dyeing, the manipulator moves the functional part S from the dye solution A to the rinsing tank for rinsing; the manipulator carries the functional part S out of the rinsing tank and puts it back to the original position of the bottle cap table; the section on the grid rack Carry out air-drying; the manipulator grabs the functional part S, removes the functional part S from the bottle cap table and moves it to the dye solution bottle B; after dyeing, the manipulator carries the functional part S from the dye solution B to the rinse tank for rinsing; the manipulator carries the functional part S was removed from the rinsing tank and returned to the original position of the cap table; the slices on the grid rack were air-d
  • the step of rinsing in the rinsing tank includes: the rinsing liquid first enters the rinsing tank chamber A from the infusion tube, and then flows from the connecting wall of the rinsing liquid chamber A and the rinsing liquid chamber B. In the several gaps of the rinsing liquid, the jet flows into the rinsing liquid chamber B in a nearly horizontal manner to realize rinsing.
  • Figure 1 is a diagram of part of the dyeing instrument. Among them, 1 is a support platform, 2 is a support column, 3 is a bottle cap platform, 4 is a rinsing tank, 5 is a dyeing bottle A, 6 is a dyeing bottle B, 7 is a waste liquid tank, and 8 is a functional part S.
  • Fig. 2 is a part diagram of the bottle cap platform.
  • FIG. 3 is a diagram of parts of the rinse tank. 41 is the rinse liquid chamber A, 42 is the rinse liquid inlet pipe interface, 43 is the connecting wall, 44 is the rinse liquid chamber B, and 45 is the waste liquid outlet.
  • Figure 4 is an overall view of the staining instrument. Wherein, 9 is a manipulator; 10 is a stepping motor x, 11 is a stepping motor y, and 12 is a stepping motor z.
  • Transfer high-throughput ultrathin section stainer includes:
  • Supporting platform is not limited to the circular shape in the accompanying drawings, and can also be set in a triangular, quadrangular, pentagonal or more than five-sided shape according to specific needs.
  • Support column described support column can be one or more, if support column selects one, then support column is preferably in the center position of support platform and is fixed on the support platform, and the height of described support column will be higher than the maximum Height and ensure that the grid frame is stable and suspended without contact with other parts;
  • Bottle cap platform the center position of the bottle cap platform is fixed on the other end of the supporting column; the corresponding number of stations is set on the bottle cap platform according to the needs of the functional parts S, the rinsing tank and the dyeing bottle, in this embodiment Taking three stations as an example, the first station is a hollow hole, which can hold the dyeing functional part S, and the top of the functional part S can be designed with the size and shape of the caps of the dye solution bottles A and B.
  • the lower end can be clamped and uploaded to the grid frame; the second station and the third station match the size and size of the caps of dye solution bottles A and B.
  • Rinse pool, staining bottle A and staining bottle B, described rinsing pool, staining bottle A and staining bottle B are all fixed on the support platform and around the support column; the size of the rinsing pool, staining bottle A and staining bottle B It matches the size and shape of the grid rack of the shape and functional part S, and is used to dye or rinse the grid of the grid rack; if the horizontal width of the grid rack is greater than the vertical height, the rinsing tank, The size and shape of dyeing bottle A and dyeing bottle B should be set accordingly so that the horizontal width is greater than the vertical height; The size and shape of B should be set so that the vertical height is greater than the horizontal width.
  • the rinsing tank includes a rinsing liquid chamber A, a rinsing liquid inlet pipe interface, a connecting wall, a rinsing liquid chamber B, and a waste liquid outlet; one side of the rinsing liquid chamber A has a rinsing liquid inlet pipe interface, and the other One side is connected to the rinse liquid chamber B through a connecting wall; there is a waste liquid outlet at the bottom of the rinse liquid chamber B; the connecting wall has a number of small holes or slits, and the liquid introduced from the rinse liquid chamber A is Enter the rinsing liquid chamber B through the connecting wall at a certain angle or in a direction close to parallel, so that the rinsing liquid can be rinsed in a direction nearly perpendicular to
  • Dyeing bottle A and dyeing bottle B are used to hold the dyeing liquid and realize the dyeing of the carrier grid.
  • This embodiment is set with two kinds of dyeing liquids. More than three staining liquids can be correspondingly arranged as three or more staining bottles according to the scheme of this embodiment.
  • a waste liquid pool, the waste liquid pool is below the support platform, and is used to receive the liquid flowing out from the waste liquid outlet of the rinsing pool.
  • Manipulator and 3 stepper motors the manipulator and 3 stepper motors are arranged above the support platform, and 3 stepper motors control the movement of the manipulator in the three directions of X, Y, and Z, so as to realize the movement of the manipulator to the object Grab and move; the manipulator is used to grab and place the caps of dye solution bottle A and dye solution bottle B, and realize the operation of capping and uncapping; The action of grabbing and grabbing the top cover of the functional parts of the grid; the stepping motor is arranged on the keel frame.
  • the air-drying system includes four-corner and top fans, and the air-drying system is arranged around the supporting platform, and is used for air-drying the slices carried by the net frame.
  • a pipeline and a flow meter are matched with the rinsing tank and control the flow and velocity of the liquid in the rinsing tank.
  • the gas-phase fluid system includes a four-corner fan system that matches the function of the air-drying grid carrier and the air outlet on the top of the box.
  • the stepping motor controls the manipulator to move to the top of the dye solution bottle A, the manipulator grabs the bottle cap A' on the dye solution bottle A to leave the dye solution bottle, and the A cover is placed on the bottle cap table.
  • the manipulator grabs the functional component S and enters the dye solution A under the control of the stepping motor.
  • the manipulator moves the functional part S from the dye solution A to the position of the rinsing tank under the control of the stepping motor.
  • the manipulator releases the top cover of the functional part S, moves to the bottle cap A', grabs the bottle cap A', moves to the top of the dye bottle A under the control of the stepping motor, releases the bottle cap A', and After the dyeing bottle A is sealed, return to the starting position.
  • the manipulator takes the functional part S out of the rinsing tank and puts it back to the original position of the bottle cap table.
  • the manipulator grabs the functional component S under the control of the stepping motor, and moves the functional component S out of the bottle cap table and into the dye solution bottle B.
  • the device of the invention can automatically perform dyeing, rinsing, air-drying, re-dyeing, rinsing, and air-drying of batches of nets inserted on the net rack, thereby improving work efficiency and saving dyeing liquid.

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Abstract

本发明属于透射电镜技术领域。本发明提供了一种转移式高通量超薄切片染色仪，包括：支撑平台；支撑柱，所述支撑柱在支撑平台中心位置并固定在支撑平台上；瓶盖平台，所述瓶盖平台的中心位置固定在支撑柱的另一端；漂洗池、染色瓶A和染色瓶B,所述漂洗池、染色瓶A和染色瓶B都固定在支撑平台上并在支撑柱周围。本发明的装置能对载网架上插放的批量载网进行染液染色、漂洗、风干、再染液染色、漂洗、风干的自动化操作，提高工作效率，节约染液。

Description

转移式高通量超薄切片染色仪 技术领域
本发明属于透射电镜技术领域。
背景技术
利用透射电镜观察生物样品细胞内部超微结构时，需要利用超薄切片制样技术，将样品制成100nm厚度以内的超薄切片，并承载在一个直径约3mm、内径约为2.6nm、环宽约为0.2mm、厚度为0.02mm的载网上，而后利用镊子尽可能夹取载网的环宽位置(最大程度避免对生物切片的机械损伤)，进行染色的操作后，方能进行显微成像观察。该染色环节需要前后两种染液的染色，每种染液染色完成后，均需要漂洗和风干的环节，总共6步操作。
如何解决复杂手工染色问题，一直是载网染色技术的难点和痛点。
发明内容
有鉴于此，本发明提供的染色仪以发明专利ZL 201610366342.4和实用新型ZL 202020027034.0两种对称夹持型批量载网架为核心功能部件，在此基础上搭建了机械转移的工作模式，完成对载网架上插放的批量载网进行染液染色、漂洗、风干、再染液染色、漂洗、风干的自动化操作。
本发明提供了一种转移式高通量超薄切片染色仪，包括：支撑平台；支撑柱，所述支撑柱在支撑平台中心位置并固定在支撑平台上；瓶盖平台，所述瓶盖平台的中心位置固定在支撑柱的另一端；漂洗池、染色瓶A和染色瓶B,所述漂洗池、染色瓶A和染色瓶B都固定在支撑平台上并在支撑柱周围。
在本发明的具体实施方式中，所述瓶盖平台上有三个工位，其中第一工位为中空孔位，第二工位和第三工位与染液瓶A和B的瓶盖的大小和尺寸相匹配。
在本发明的具体实施方式中，所述漂洗池包括：漂洗液腔室A、漂洗液进液管接口、连接壁、漂洗液腔室B、废液出口；所述漂洗液腔室A的一侧有漂洗液进液管接口，另一侧通过连接壁与漂洗液腔室B相连；所述漂洗液腔室B底部有废液出口；所述连接壁有若干孔径细小的孔洞或者狭缝，从漂洗液腔室A引入的液体接近平行方向通过连接壁进入漂洗液腔室B,达到漂洗液以近乎与载网架载网垂直的方向对其进行漂洗；所述废液出口与废液池相同并实现废液流入废液池。
在本发明的具体实施方式中，还包括废液池，所述废液池在支撑平台下方，用于接收漂洗池废液出口流出的液体。
在本发明的具体实施方式中，还包括：机械手和3个步进电机；所述机械手和3个步进电机设置在支撑平台上方，有3个步进电机控制控制机械手在X、Y、Z三个方向的移动，进而实现机械手对物品的抓取和运动；所述机械手用来抓取和抓放染液瓶A和染液瓶B的瓶盖， 并实现盖盖和拔盖的操作；所述机械手还可以对已装卡上的载网架的功能部件的顶盖的抓取和抓放的动作；所述步进电机设置在龙骨架上。
在本发明的具体实施方式中，所述风干系统，所述风干系统包括四角和顶端风扇。
本发明还提供了一种转移式高通量超薄切片染色仪的染色方法，包括如下步骤：机械手抓取染液瓶A上的瓶盖A’离开染液瓶；机械手抓取功能部件S进入染液A中；染色后机械手携功能部件S从染液A中移动到漂洗池中漂洗中；机械手携功能部件S从漂洗池中移出并放回瓶盖台原位；将载网架上的切片进行风干；机械手抓取功能部件S，携功能部件S从瓶盖台中移出并移动到染液瓶B中；染色后机械手携功能部件S从染液B中移动到漂洗池中漂洗；机械手携功能部件S从漂洗池中移出并放回瓶盖台原位；将载网架上的切片进行风干。
在本发明的具体实施方式中，所述漂洗池中漂洗的步骤包括：漂洗液从输液管中先进入漂洗池腔室A中，而后从漂洗液腔室A和漂洗液腔室B的连接壁的若干空隙中以近乎水平方式喷流到漂洗液腔室B中实现漂洗。
附图说明
图1是染色仪部分装置图。其中，1是支撑平台，2是支撑柱，3是瓶盖平台，4是漂洗池，5是染色瓶A，6是染色瓶B，7是废液池，8是功能部件S。
图2是瓶盖平台部件图。
图3是漂洗池部件图。其中41是漂洗液腔室A，42是漂洗液进液管接口，43是连接壁，44是漂洗液腔室B，45是废液出口。
图4是染色仪整体图。其中，9是机械手；10是步进电机x，11是步进电机y，12是步进电机z。
具体实施方式
实施例1
转移式高通量超薄切片染色仪包括：
支撑平台；所述支撑平台不限于附图中圆形，也可以依据具体需要设置成三边形、四边形、五边形及五边以上形状等。
支撑柱，所述支撑柱可以为一个或多个，若支撑柱选择一个，则支撑柱优选在支撑平台中心位置并固定在支撑平台上，所述支撑柱的高度要高于载网架的最大高度并保证载网架稳定悬空且不与其他部件接触；
瓶盖平台，所述瓶盖平台的中心位置固定在支撑柱的另一端；所述瓶盖平台上依据功能部件S、漂洗池和染色瓶的需要设置成相应数量的工位，本实施例中以设有三个工位为例，其中第一工位为中空孔位，可卡套住染色功能部件S，所述功能部件S的顶端为与染液瓶A和B瓶盖大小、形状可以设计为一致的瓶盖结构或不一致的结构，下端可装卡上载网架；第二工位和第三工位与染液瓶A和B的瓶盖的大小和尺寸相匹配。
漂洗池、染色瓶A和染色瓶B,所述漂洗池、染色瓶A和染色瓶B都固定在支撑平台上并在支撑柱周围；所述漂洗池、染色瓶A和染色瓶B的大小尺寸和形状与功能部件S的载网架的尺寸和形状相匹配，用于对载网架的载网进行染色或漂洗；如依据载网架的横向的宽度大于竖向的高度，则漂洗池、染色瓶A和染色瓶B的尺寸和形状要相应设置为横向的宽度大于竖向的高度；或者如依据载网架的竖向的高度大于横向的宽度，则漂洗池、染色瓶A和染色瓶B的尺寸和形状要相应设置为竖向的高度大于横向的宽度。
漂洗池，所述漂洗池下端有液体出口或管道并与废液池相通；所述漂洗池用于对载网架载网实现漂洗。所述漂洗池包括漂洗液腔室A、漂洗液进液管接口、连接壁、漂洗液腔室B、废液出口；所述漂洗液腔室A的一侧有漂洗液进液管接口，另一侧通过连接壁与漂洗液腔室B相连；所述漂洗液腔室B底部有废液出口；所述连接壁有若干孔径细小的孔洞或者狭缝，从漂洗液腔室A引入的液体以一定角度或接近平行方向通过连接壁进入漂洗液腔室B,达到漂洗液以近乎与载网架载网垂直的方向或成一定角度对其进行漂洗；所述废液出口与废液池相同并实现废液流入废液池。
染色瓶A和染色瓶B，所述染色瓶A和染色瓶B用于盛放染色液体并对载网架载网实现染色，本实施例以两种染色液需要而设置，如若需要三个或三个以上染色液，可以依据本实施例方案相应设置成三个或三个以上染色瓶。
废液池，所述废液池在支撑平台下方，用于接收漂洗池废液出口 流出的液体。
机械手和3个步进电机，所述机械手和3个步进电机设置在支撑平台上方，有3个步进电机控制控制机械手在X、Y、Z三个方向的移动，进而实现机械手对物品的抓取和运动；所述机械手用来抓取和抓放染液瓶A和染液瓶B的瓶盖，并实现盖盖和拔盖的操作；所述机械手还可以对已装卡上的载网架的功能部件的顶盖的抓取和抓放的动作；所述步进电机设置在龙骨架上。
风干系统，包括四角和顶端风扇，所述风干系统设置在支撑平台周围，用于对载网架载网的切片进行风干。
管路和流量计，所述管路和流量计与漂洗池相配套并控制漂洗池液体的流量和流速。
5)气相流体系统包括与风干载网架功能相配套的四角风扇系统及箱体顶端的出风口。
转移式高通量超薄切片染色仪的工作方法，以两种染色液为例：
(1)、将染液瓶A、B(携对应瓶盖A’、B’)放置在相应工位；将装卡载网架放在瓶盖台上功能部件S的工位。
(2)、步进电机控制机械手移动到染液瓶A上方，机械手抓取染液瓶A上的瓶盖A’离开染液瓶，A盖放在瓶盖台上。
(3)、机械手抓取功能部件S在步进电机控制下进入染液A中。
(4)、染色一定时间后，机械手携功能部件S在步进电机控制下从染液A中移动到漂洗池位置。
(5)、①漂洗池的流量阀启动工作，漂洗液从输液管中先进入漂洗池腔室A中，而后从漂洗液腔室A和漂洗液腔室B的连接壁的若干空隙中以近乎水平方式或成一定角度喷流到漂洗液腔室B中实现漂洗，废液从漂洗液腔室B下端的废液出口流入废液池。
②于此同时，机械手松开功部件S顶盖，移动到瓶盖A’处，抓取瓶盖A’，在步进电机控制下移动到染液瓶A上方，释放瓶盖A’，将染色瓶A密封后，回到起始位。
(6)、漂洗池的流量阀停止漂洗液继续进入。
(7)、机械手携功能部件S从漂洗池中移出并放回瓶盖台原位。
(8)、四角风扇启动，对载网架上的切片进行风干。
(9)、机械手在步进电机控制下，抓取染液瓶B中瓶盖B’，并在步进电机控制下将瓶盖B’放置在瓶盖台。
(10)、机械手在步进电机控制下抓取功能部件S，携功能部件S从瓶盖台中移出并移动到染液瓶B中。
(11)、染色一段时间后对功能部件S和对应的染液瓶盖B’重复相应4-8步骤。
本发明的装置能对载网架上插放的批量载网进行染液染色、漂洗、风干、再染液染色、漂洗、风干的自动化操作，提高工作效率，节约染液。

Claims (9)

  1. 转移式高通量超薄切片染色仪，其特征在于，包括：
    支撑平台；
    支撑柱，所述支撑柱在支撑平台中心位置并固定在支撑平台上；
    瓶盖平台，所述瓶盖平台的中心位置固定在支撑柱的另一端；
    漂洗池、染色瓶A和染色瓶B,所述漂洗池、染色瓶A和染色瓶B都固定在支撑平台上并在支撑柱周围。
  2. 依据权利要求1所述转移式高通量超薄切片染色仪，其特征在于，所述瓶盖平台上有三个工位，其中第一工位为中空孔位，第二工位和第三工位与染液瓶A和B的瓶盖的大小和尺寸相匹配。
  3. 依据权利要求1所述转移式高通量超薄切片染色仪，其特征在于，所述漂洗池包括：
    漂洗液腔室A、漂洗液进液管接口、连接壁、漂洗液腔室B、废液出口；
    所述漂洗液腔室A的一侧有漂洗液进液管接口，另一侧通过连接壁与漂洗液腔室B相连；
    所述漂洗液腔室B底部有废液出口；
    所述连接壁有若干孔径细小的孔洞或者狭缝，从漂洗液腔室A引入的液体以一定角度或接近平行方向通过连接壁进入漂洗液腔室B,达到漂洗液以近乎与载网架载网垂直的方向或成一定角度对其进行漂洗；
    所述废液出口与废液池相同并实现废液流入废液池。
  4. 依据权利要求1所述转移式高通量超薄切片染色仪，其特征在于，所述漂洗池、染色瓶A和染色瓶B的竖向的高度大于横向的宽度或者横向的宽度大于竖向的高度。
  5. 依据权利要求1至4任一所述转移式高通量超薄切片染色仪，其特征在于，还包括废液池，所述废液池在支撑平台下方，用于接收漂洗池废液出口流出的液体。
  6. 依据权利要求1至4任一所述转移式高通量超薄切片染色仪，其特征在于，还包括：
    机械手和3个步进电机；
    所述机械手和3个步进电机设置在支撑平台上方，有3个步进电机控制控制机械手在X、Y、Z三个方向的移动，进而实现机械手对物品的抓取和运动；
    所述机械手用来抓取和抓放液瓶A和染液瓶B的瓶盖，并实现盖盖和拔盖的操作；
    所述机械手还可以对已装卡上的载网架的功能部件的顶盖的抓取和抓放的动作；
    所述步进电机设置在龙骨架上。
  7. 依据权利要求1至4任一所述转移式高通量超薄切片染色仪，其特征在于，所述风干系统，所述风干系统包括四角和顶端风扇。
  8. 依据权利要求1至4任一所述转移式高通量超薄切片染色仪的染色方法，其特征在于，包括如下步骤：
    机械手抓取染液瓶A上的瓶盖A’离开染液瓶；
    机械手抓取功能部件S进入染液A中；
    染色后机械手携功能部件S从染液A中移动到漂洗池中漂洗中；
    机械手携功能部件S从漂洗池中移出并放回瓶盖台原位；
    将载网架上的切片进行风干；
    机械手抓取功能部件S，携功能部件S从瓶盖台中移出并移动到染液瓶B中；
    染色后机械手携功能部件S从染液B中移动到漂洗池中漂洗；
    机械手携功能部件S从漂洗池中移出并放回瓶盖台原位；
    将载网架上的切片进行风干。
  9. 依据权利要求8所述转移式高通量超薄切片染色仪的染色方法，其特征在于，所述漂洗池中漂洗的步骤包括：
    漂洗液从输液管中先进入漂洗池腔室A中，而后从漂洗液腔室A和漂洗液腔室B的连接壁的若干空隙中以近乎水平方式或以一定角度喷流到漂洗液腔室B中实现漂洗。
PCT/CN2022/125798 2021-11-22 2022-10-18 转移式高通量超薄切片染色仪 WO2023088014A1 (zh)

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