Transfer-type high-throughput ultrathin section staining instrument

Abstract

The present invention belongs to the technical field of transmission electron microscopy. The present invention provides a transfer-type high-throughput ultrathin section staining instrument, comprising: a support platform; a support column, the support column is located at the center of the support platform and fixed on the support platform; a bottle cap platform, the center of the bottle cap platform is fixed at the other end of the support column; a rinsing tank, a staining bottle A and a staining bottle B, the rinsing tank, the staining bottle A and the staining bottle B are all fixed on the support platform and around the support column. The device of the present invention can automatically perform dyeing, rinsing, air drying, dyeing again, rinsing, and air drying on a batch of carrier nets inserted on a carrier net rack, thereby improving work efficiency and saving dyeing liquid.

Description

Technical Field

[0001] The invention belongs to the technical field of transmission electron microscopes. Background Art

[0002] When using a transmission electron microscope to observe the internal ultrastructure of biological sample cells, it is necessary to use ultrathin sectioning technology to make the sample into ultrathin sections with a thickness of less than 100nm, and carry it on a grid with a diameter of about 3mm, an inner diameter of about 2.6nm, a ring width of about 0.2mm, and a thickness of 0.02mm. Then use tweezers to clamp the ring width position of the grid as much as possible (to minimize mechanical damage to the biological slices), and perform the staining operation before microscopic imaging observation. This staining process requires staining with two dye solutions, and after each dye solution staining is completed, it is necessary to rinse and air dry, a total of 6 steps.

[0003] How to solve the problem of complex manual dyeing has always been the difficulty and pain point of screen dyeing technology. Summary of the invention

[0004] In view of this, the dyeing instrument provided by the present invention takes two symmetrical clamping type batch grid racks of invention patent ZL 201610366342.4 and utility model ZL202020027034.0 as core functional components, and builds a mechanical transfer working mode on this basis to complete the automated operations of dyeing with dye liquid, rinsing, air-drying, re-dyeing with dye liquid, rinsing and air-drying of the batch grids inserted on the grid rack.

[0005] The present invention provides a transfer-type high-throughput ultrathin section staining instrument, comprising: a supporting platform; a supporting column, which is located at the center of the supporting platform and fixed on the supporting platform; a bottle cap platform, the center of which is fixed at the other end of the supporting column; a rinsing tank, a staining bottle A and a staining bottle B, which are all fixed on the supporting platform and around the supporting column.

[0006] In a specific embodiment of the present invention, there are three workstations on the bottle cap platform, wherein the first workstation is a hollow hole position, and the second and third workstations match the sizes and dimensions of the bottle caps of dye bottles A and B.

[0007] In a specific embodiment of the present invention, 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 is provided with a rinsing liquid inlet pipe interface, and the other side is connected to the rinsing liquid chamber B through a connecting wall; the bottom of the rinsing liquid chamber B is provided with a waste liquid outlet; the connecting wall has a number of holes or slits with small apertures, and the liquid introduced from the rinsing liquid chamber A enters the rinsing liquid chamber B through the connecting wall in a nearly parallel direction, so that the rinsing liquid rinses the grid rack in a direction nearly perpendicular to the grid rack; the waste liquid outlet is the same as the waste liquid tank and enables the waste liquid to flow into the waste liquid tank.

[0008] In a specific embodiment of the present invention, a waste liquid pool is further included. The waste liquid pool is below the supporting platform and is used to receive liquid flowing out of the waste liquid outlet of the rinsing tank.

[0009] In a specific embodiment of the present invention, it also includes: a manipulator and three stepper motors; the manipulator and the three stepper motors are arranged above the supporting platform, and the three stepper motors control the movement of the manipulator in the three directions of X, Y and Z, thereby realizing the manipulator's grasping and movement of objects; the manipulator is used to grasp and place the bottle caps of dye bottle A and dye bottle B, and realize the operations of capping and removing the caps; the manipulator can also grasp and place the top covers of the functional components of the carrier grid that have been clamped; the stepper motor is arranged on the keel frame.

[0010] In a specific embodiment of the present invention, the air-drying system includes four-corner and top fans.

[0011] The present invention also provides a staining method for a transfer-type high-throughput ultrathin section staining instrument, comprising the following steps: a robot grabs a bottle cap A' on a dye bottle A and leaves the dye bottle; the robot grabs a functional component S and enters the dye A; after staining, the robot carries the functional component S and moves it from the dye A to a rinsing tank for rinsing; the robot carries the functional component S out of the rinsing tank and puts it back to the original position of the bottle cap platform; the sections on the grid rack are air-dried; the robot grabs the functional component S, carries the functional component S out of the bottle cap platform and moves it to a dye bottle B; after staining, the robot carries the functional component S and moves it from the dye B to the rinsing tank for rinsing; the robot carries the functional component S out of the rinsing tank and puts it back to the original position of the bottle cap platform; the sections on the grid rack are air-dried.

[0012] In a specific embodiment of the present invention, the step of rinsing in the rinsing pool includes: the rinsing liquid first enters the rinsing pool chamber A from the infusion tube, and then sprays into the rinsing liquid chamber B in a nearly horizontal manner from several gaps in the connecting wall of the rinsing liquid chamber A and the rinsing liquid chamber B to achieve rinsing. BRIEF DESCRIPTION OF THE DRAWINGS

[0013] Figure 1This is a partial device diagram of the dyeing instrument. Among them, 1 is the support platform, 2 is the support column, 3 is the bottle cap platform, 4 is the rinsing tank, 5 is the dyeing bottle A, 6 is the dyeing bottle B, 7 is the waste liquid tank, and 8 is the functional component S.

[0014] Figure 2 This is a diagram of the bottle cap platform components.

[0015] Figure 3 41 is a rinse liquid chamber A, 42 is a rinse liquid inlet pipe interface, 43 is a connecting wall, 44 is a rinse liquid chamber B, and 45 is a waste liquid outlet.

[0016] Figure 4 9 is a schematic diagram of the dyeing apparatus, wherein 9 is a manipulator, 10 is a stepper motor x, 11 is a stepper motor y, and 12 is a stepper motor z. DETAILED DESCRIPTION

[0017] Example 1

[0018] The transfer-type high-throughput ultrathin section stainer includes:

[0019] Support platform;

[0020] A support column, which is located at the center of the support platform and fixed on the support platform. The height of the support column is higher than the maximum height of the grid frame and ensures that the grid frame is stably suspended and does not contact other components;

[0021] A bottle cap platform, the center position of which is fixed at the other end of the support column; there are three workstations on the bottle cap platform, wherein the first workstation is a hollow hole position, which can be used to clamp the dyeing functional component S, the top of which is a bottle cap structure that is consistent with the size and shape of the bottle caps of dye bottles A and B, and the lower end can be clamped on the loading grid; the second and third workstations are matched with the size and dimensions of the bottle caps of dye bottles A and B.

[0022] The rinsing tank, the dyeing bottle A and the dyeing bottle B are all fixed on the supporting platform and around the supporting column.

[0023] A rinsing tank, wherein the lower end of the rinsing tank has a liquid outlet or a pipe and is connected to a waste liquid tank; the rinsing tank is used to rinse the grid carrier. 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 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 holes or slits with small apertures, and the liquid introduced from the rinsing liquid chamber A enters the rinsing liquid chamber B through the connecting wall in a nearly parallel direction, so that the rinsing liquid rinses the grid carrier in a direction nearly perpendicular to the grid carrier; the waste liquid outlet is the same as the waste liquid tank and enables the waste liquid to flow into the waste liquid tank.

[0024] The dyeing bottle A and the dyeing bottle B are used to contain dyeing liquid and dye the grid carrier.

[0025] The waste liquid pool is located below the supporting platform and is used to receive liquid flowing out of the waste liquid outlet of the rinsing pool.

[0026] A manipulator and three stepper motors are arranged above the supporting platform, and the three stepper motors control the movement of the manipulator in three directions of X, Y and Z, so as to realize the grabbing and movement of objects by the manipulator; the manipulator is used to grab and place the bottle caps of dye bottle A and dye bottle B, and realize the operations of capping and removing the caps; the manipulator can also grab and place the top covers of the functional components of the carrier grid that have been installed; the stepper motor is arranged on the keel frame.

[0027] The air drying system comprises fans at four corners and a top end, and the air drying system is arranged around the supporting platform and is used for air drying the slices of the grid carrier.

[0028] Pipes and flow meters are matched with the rinsing tank and control the flow rate and flow velocity of the liquid in the rinsing tank.

[0029] 5) The gas phase fluid system includes a four-corner fan system that matches the air-drying grid frame function and an air outlet at the top of the box.

[0030] Working method of transfer high-throughput ultra-thin section staining instrument:

[0031] (1) Place dye bottles A and B (with corresponding bottle caps A' and B') at the corresponding workstations; place the card carrier rack at the workstation of functional component S on the bottle cap platform.

[0032] (2) The stepper motor controls the manipulator to move to the top of the dye bottle A. The manipulator grabs the bottle cap A' on the dye bottle A and leaves the dye bottle. The cap A is placed on the bottle cap stand.

[0033] (3) The robot grabs the functional component S and enters the dye solution A under the control of the stepper motor.

[0034] (4) After dyeing for a certain period of time, the robot carrying the functional component S moves from the dye solution A to the rinsing tank under the control of the stepper motor.

[0035] (5) ① The flow valve of the rinsing tank is started, and the rinsing liquid first enters the rinsing tank chamber A from the infusion tube, and then sprays into the rinsing liquid chamber B in a nearly horizontal manner from the several gaps in the connecting wall of the rinsing liquid chamber A and the rinsing liquid chamber B to achieve rinsing, and the waste liquid flows into the waste liquid tank from the waste liquid outlet at the lower end of the rinsing liquid chamber B.

[0036] ②At the same time, the robot releases the top cover of the working component 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 stepper motor, releases the bottle cap A', seals the dye bottle A, and returns to the starting position.

[0037] (6) The flow valve of the rinse tank stops the rinse liquid from entering.

[0038] (7) The robot carries the functional component S out of the rinsing tank and puts it back to its original position on the bottle cap table.

[0039] (8) Start the fans at the four corners to air dry the slices on the grid rack.

[0040] (9) The robot arm, under the control of the stepper motor, grabs the bottle cap B' in the dye bottle B and places the bottle cap B' on the bottle cap table under the control of the stepper motor.

[0041] (10) The manipulator grabs the functional component S under the control of the stepper motor, moves the functional component S out of the bottle cap platform and moves it into the dye bottle B.

[0042] (11) After dyeing for a period of time, repeat steps 4-8 for the functional component S and the corresponding dye bottle cap B'.

[0043] The device of the invention can automatically perform dyeing, rinsing, air-drying, re-dyeing, rinsing and air-drying on batches of carrier nets inserted on the carrier net rack, thereby improving work efficiency and saving dyeing liquid.

Claims

1. A transfer-type high-throughput ultrathin section staining instrument, characterized in that: include: Support platform; A support column, wherein the support column is located at the center of the support platform and is fixed on the support platform; A bottle cap platform, the center of which is fixed at the other end of the support column; the bottle cap platform has three workstations, wherein the first workstation is a hollow hole, and the second and third workstations match the size and dimensions of the bottle caps of dye bottles A and B; A rinsing tank, a dyeing bottle A and a dyeing bottle B, wherein the rinsing tank, the dyeing bottle A and the dyeing bottle B are all fixed on the supporting platform and around the supporting column; the rinsing tank comprises: Rinsing liquid chamber A, rinsing liquid inlet pipe interface, connecting wall, rinsing liquid chamber B, waste liquid outlet; One side of the rinsing liquid chamber A is provided with a rinsing liquid inlet pipe interface, and the other side is connected to the rinsing liquid chamber B through a connecting wall; The bottom of the rinsing liquid chamber B is provided with a waste liquid outlet; The connecting wall has a plurality of holes or slits with small apertures, and the liquid introduced from the rinsing liquid chamber A passes through the connecting wall in a nearly parallel direction into the rinsing liquid chamber B, so that the rinsing liquid rinses the grid carrier in a direction nearly perpendicular to the grid carrier. The waste liquid outlet is the same as the waste liquid pool and enables the waste liquid to flow into the waste liquid pool.

2. The transfer-type high-throughput ultrathin section staining instrument according to claim 1, characterized in that: It also includes a waste liquid pool, which is located below the supporting platform and is used to receive liquid flowing out of the waste liquid outlet of the rinsing pool.

3. The transfer-type high-throughput ultrathin section staining instrument according to claim 1, characterized in that: Also includes: Robot and 3 stepper motors; The manipulator and three stepper motors are arranged above the support platform, and the three stepper motors control the movement of the manipulator in the three directions of X, Y and Z, thereby realizing the grasping and movement of the manipulator to the object; The manipulator is used to grab and place the bottle caps of the liquid bottle A and the dye bottle B, and to perform the capping and uncapping operations; The manipulator can also grasp and place the top cover of the functional component of the grid carrier that has been installed; The stepping motor is arranged on the keel frame.

4. The transfer-type high-throughput ultrathin section staining instrument according to claim 1, characterized in that: Also included is an air drying system including four corner and top fans.

5. The staining method of the transfer-type high-throughput ultrathin section staining instrument according to any one of claims 1 to 4, characterized in that: The steps include: The robot grabs the bottle cap A' on the dye bottle A and leaves the dye bottle; The robot grabs the functional component S and puts it into the dye solution A; After dyeing, the robot carries the functional component S from the dye solution A to the rinsing tank for rinsing; The robot carries the functional component S out of the rinsing tank and puts it back to the bottle cap table; Air-dry the slices on the grid rack; The robot grabs the functional component S, moves the functional component S out of the bottle cap table and moves it into the dye bottle B; After dyeing, the robot carries the functional component S from the dye solution B to the rinsing tank for rinsing; The robot carries the functional component S out of the rinsing tank and puts it back to the bottle cap table; Air dry the sections on the grid rack.

6. The staining method of the transfer-type high-throughput ultrathin section staining instrument according to claim 5, characterized in that: The step of rinsing in the rinsing tank comprises: The rinsing liquid first enters the rinsing tank chamber A from the infusion tube, and then sprays into the rinsing liquid chamber B in a nearly horizontal manner from several gaps in the connecting wall of the rinsing liquid chamber A and the rinsing liquid chamber B to achieve rinsing.

Images