CN109143563B

CN109143563B - micro-motion tracking system

Info

Publication number: CN109143563B
Application number: CN201811054472.XA
Authority: CN
Inventors: 吴旭; 曹云霞; 刘雅静; 刘振邦; 孙红娟
Original assignee: University of Science and Technology of China USTC
Current assignee: University of Science and Technology of China USTC
Priority date: 2018-09-11
Filing date: 2018-09-11
Publication date: 2020-01-31
Anticipated expiration: 2038-09-11
Also published as: CN109143563A

Abstract

The invention discloses an microscopic motion tracking system, which comprises a base, a front vertical plate, a rear vertical plate, a transverse plate, a sample fixing device, an optical imaging main body and a X, Y, Z triaxial motion mechanism, wherein a method of sample immobilization and light path system movement is adopted, so that the interference of shaking of a culture solution caused by a moving objective table to microscopic imaging is avoided under a suspension environment, a camera is used for carrying out characteristic identification and position identification on a sample image under a field of view and marking the characteristics of each micro organism living bodies, the tracking of micro organisms can be triggered according to the conditions set by a user, and the micro organisms are stabilized to form an image, and a micro operation worktable linked with a light path system can be provided with a micro operation and operate a specific target sample in a relatively static state.

Description

micro-motion tracking system

Technical Field

The invention relates to the technical field of microscopic imaging, in particular to microscopic motion tracking systems, which are systems capable of tracking large-range motion samples under a microscope.

Background

For micro organisms capable of moving in a large range in a culture solution, such as zebrafish, nematodes, plankton, sperm and the like, an imaging range is small because an objective lens of a microscope is in an enlarged state, and the micro organisms can easily move out of an observation range. Conventionally, to achieve continuous observation under a microscope, the stage must be moved. However, acceleration and deceleration of the stage movement can result in shaking of the culture fluid, which is generally unacceptable under microscopic observation conditions.

The current solution to the above-mentioned liquid shaking is to use kinds of special containers on an inverted microscope, the container has a double-glass observation chamber, the sample is injected between the double-glass, and the container can be placed under the microscope for observation, and the method has the defect that the sample can not be operated, such as catching, injecting, etc.

In addition, when the conventional microscope adjusts the imaging focal length, two knobs of coarse adjustment and fine adjustment are usually designed for target searching and fine adjustment of the focal plane, respectively, and when the microscope system is automatically designed or modified, the adjustment of the focal length is usually completed by adopting a screw rod and a motor. In order to ensure the fineness of fine adjustment, the screw pitch of the screw rod and the step angle of the motor have high requirements, or piezoelectric ceramic driving is adopted to obtain higher precision. Above-mentioned scheme manufacturing cost is high for when the repacking, has the big problem of the repacking degree of difficulty.

Disclosure of Invention

The invention aims to realize stable tracking observation and operation on tiny organisms in a suspension environment, such as zebra fish, nematodes, plankton, sperms and the like, and avoid shaking of a liquid environment due to acceleration change. According to the invention, the optical imaging mechanism is moved by fixing the objective table, so that the shaking of the culture solution can be avoided; realizing automatic focusing through image definition identification; a differential gear reducer is designed to replace a manual focusing mechanism and simultaneously serve as an actuator for automatic focusing. The specific micro organism is identified and tracked through the image processing and analyzing technology, so that the stable imaging is realized, and the specific micro organism is always positioned in the observation range, thereby facilitating the long-time physiological state observation and operation of the specific target.

The technical scheme includes that the microscopic motion tracking system comprises an equipment base, a front vertical plate, a rear vertical plate, a sample fixing device and a transverse plate, wherein the equipment base, the front vertical plate, the rear vertical plate and the transverse plate are fixedly connected through bolts to form stable whole bodies, the sample fixing device is fixed in an opening of the transverse plate and is kept stable and fixed, a Y-direction moving base is connected with the equipment base through 2Y-direction linear guide rails, a Y-direction driving motor and a ball screw rod to form a , meanwhile, an X-direction moving base is fixed to through 2 triangular supports and the Y-direction moving base, an upper light path mounting support plate and a lower light path mounting support plate are connected to through 2X-direction linear guide rails, the X-direction driving motor and the ball screw rod, the upper light path system is mounted in front of the upper light path mounting support plate and the lower light path mounting support plate, the lower light path system is mounted above the upper light path mounting support plate and the lower light path mounting support plate, the two micro-operated devices are also mounted above the upper light path mounting support plate and the lower light path mounting plate, the upper light path system comprises a light source, a reflector, a condenser lens, a.

The whole upper and lower light path system and the micro-operation device are connected into bodies and can move in any direction in a plane under the drive of X, Y motor.

The objective lens focusing mechanism can be a manual focusing mechanism, an electric focusing mechanism in a lead screw guide rail mode, or an electric focusing mechanism based on a differential gear reducer.

The electric focusing mechanism of the differential gear reducer is small in size, strong in adaptability and suitable for automatic modification on the basis of a manual focusing mechanism. The function of the advanced microscope can be realized at low cost by scientific research personnel.

Compared with the prior art, the invention has the advantages that:

the invention can greatly reduce the difficulty of operating living micro organisms, improve the working efficiency, reduce the workload of experimenters, enable professional operators to get rid of long-term training and physiological challenges, reduce the dependence on the professional experimenters, realize the high-difficulty microscopic operation which is difficult to be reached by manual work, and provide a new auxiliary research tool for biomedical engineering. The technical scheme provided by the invention can be realized by modifying a conventional microscope.

Drawings

FIG. 1 is an exploded view of the structure of the system of the present invention;

FIG. 2 is a view of the construction of the sample holding device of the present invention;

FIG. 3 is a front mounting schematic view of the Y-motion mechanism of the present invention;

FIG. 4 is a rear mounting schematic view of the Y-motion mechanism of the present invention;

FIG. 5 is a front mounting schematic view of the X-direction movement mechanism of the present invention;

FIG. 6 is a rear mounting schematic view of the X-direction movement mechanism of the present invention;

FIG. 7 is a schematic view of the mounting of the Z-focus mechanism of the present invention;

FIG. 8 is a schematic view of the overall motion mechanism of the present invention;

FIG. 9 is a schematic view of the overall structure of the apparatus of the present invention;

FIG. 10 is an exploded schematic view of the differential transmission gear set;

FIG. 11 is a differential speed change gear set assembly schematic;

FIG. 12 is a partial assembled diagrammatic view of the differential shift gear set;

FIG. 13 is a partial assembly schematic view of the differential gear set.

In the figure: 1 equipment base, 2 front vertical plate, 3 rear vertical plate, 4 sample fixing device, 5 transverse plate, 6Y-direction driving motor and ball screw, 7Y-direction linear guide rail, 8Y-direction moving base, 9 lower light path system, 9.1 microscope focusing gearbox, 9.2 microscope focusing output gear, 9.3 microscope focusing rack, 9.4 microscope, 10 upper and lower light path mounting support plate, 11 micro-operation mounting rack, 12 micro-operation device, 13 upper light path system, 14X-direction moving base, 15X-direction driving motor and ball screw, 16X-direction linear guide rail, 101 output double gear, 102 small transmission gear, 103 transmission support cover plate, 104 output transmission shaft, 105 large transmission shaft, 106 transmission support, 107 main body frame, 108 planet carrier, 109 sun gear, 110 planet gear shaft, 111 driving pinion, 112 double planet gear, 113 planet carrier gear, 114 differential large gear, 115 motor connecting plate, 116 active stepper motor, 117 differential stepper motor.

Detailed Description

The invention is further illustrated in conjunction with the figures and the detailed description.

The microscopic motion tracking systems of the invention design stable equipment bases, wherein the front and back peripheries of the equipment bases are provided with a front vertical plate and a back vertical plate or upright post, upper panels are fixedly arranged above the front vertical plate and the back vertical plate or upright post, holes are arranged on the upper panels, and a sample fixing device (a sample clamp) is embedded and arranged in the holes of the upper panels, thus the sample can be kept stable and still.

In addition , a X, Y console is arranged above the base of the device and in the space enclosed by the front and back vertical plates and the upper panel, the whole light path system of the microscope and the micro-operation mounting platform and the micro-operation device for micro-operation are arranged on the X, Y console, the light path system and the micro-operation mounting platform are driven by a motor X, Y and a ball screw rod to move in any direction in the plane, and the light path system and the micro-operation device can be moved to the position of the sample under the condition that the sample is kept still.

The method comprises the steps of carrying out feature recognition and position recognition on a sample image in a visual field through a data acquisition camera, and identifying the features of living microorganisms, wherein the features comprise but are not limited to movement speed, shape, size, color, content and the like, and according to conditions set by a user, the tracking of living microorganisms can be triggered.

After micro-organism samples are tracked, the micro-organism samples are kept in a relatively static state and can be automatically placed in the operating range of micro-operation, so that the next steps of operation, such as suction, clamping, injury, injection and the like, are easily carried out.

In order to realize that the light path system of the microscope can track the motion track of a sample in real time and enable a microscopic image to be always aligned with the sample, the invention provides sets of motion mechanisms, and the motion tracks of cells can be automatically tracked by the light path system in the X direction and the Y direction through the motion mechanisms.

The invention solves the technical problem that two sets of Y-direction linear guide rails and a Y-direction driving motor are arranged on a base of the equipment, a Y-direction moving base of the whole moving system is connected with the Y-direction motor through the linear guide rails and the ball screw to drive the Y-direction movement of the whole system, a vertical X-direction moving base is arranged at the rear part of the base, two sets of X-direction linear guide rails and an X-direction driving motor are arranged on the vertical X-direction moving base, an upper light path system, a lower light path mounting frame and a micro-manipulation device are arranged in front of the upper light path mounting plate and the lower light path mounting plate, a Z-direction moving device is arranged in the lower light path system, a Z-direction focusing moving device can be arranged, the cell can be automatically focused, the cell can be automatically shot in real time, the cell can be tracked, and the cell can move along with the movement of the cell, and the whole cell can be driven to move along with the movement of the cell, the lower light path system, the cell can be automatically focused, and the cell can be shot in real time.

As shown in FIG. 1, the sample-fixed microscopic imaging device of the invention comprises an apparatus base 1, a front vertical plate 2, a rear vertical plate 3, a sample-fixing device 4 and a transverse plate 5.

As shown in FIG. 2, the apparatus base 1, the front vertical plate 2, the rear vertical plate 3 and the horizontal plate 5 are fastened and connected by bolts to form stable units, and the sample fixing device 4 is fixed in the opening of the horizontal plate 5 and remains stable.

As shown in fig. 3 and 4, the Y-direction moving base 8 is connected to the apparatus base 1 via 2Y-direction linear guides 7, a Y-direction driving motor, and a ball screw 6 at , and the X-direction moving base 14 is fixed to the Y-direction moving base 8 at by 2 triangular brackets.

As shown in fig. 5 and 6, the upper and lower optical path installation support plates 10 are connected to the X-direction moving base 14 through 2X-direction linear guides 16, an X-direction driving motor, and a ball screw 15 at , the upper optical path system 13 is installed above the upper and lower optical path installation support plates 10, the lower optical path system 9 is installed in front of the upper and lower optical path installation support plates 10, and two micro-manipulation mounts 11 and micro-manipulation devices 12 are also installed above the upper and lower optical path installation support plates 10.

In fig. 7, the micro-focusing gear 9.3 is driven by the micro-focusing gear box 9.1 and the micro-focusing output gear 9.2 to realize the precise movement of the microscope 9.4 on the Z axis. Thereby achieving an accurate focusing of the microscope 9.4.

In fig. 8, the whole upper and lower optical path systems and the micro-manipulation device are connected to form bodies, and can be driven by X, Y directional motors to move in any direction in a plane.

As shown in fig. 10, the differential gear reducer is used in place of the manual focus mechanism, and at the same time, serves as an actuator for automatic focusing. The differential gear reducer comprises an output duplicate gear 101, a small transmission gear 102, a transmission bracket cover plate 103, an output transmission shaft 104, a large transmission shaft 105, a transmission bracket 106, a main body frame 107, a planet carrier 108, sun teeth 109, a planet gear shaft 110, a driving pinion 111, a duplicate planet gear 112, a planet carrier gear 113, a differential large gear 114, a motor connecting plate 115, a driving stepping motor 116 and a differential stepping motor 117.

As shown in FIG. 11, the transmission bracket 106 is mounted on the main body frame 107, the output transmission shaft 104 and the large transmission shaft 105 are respectively mounted in the shaft holes of the transmission bracket 106 with bearings, the transmission bracket cover plate 103 is mounted on the transmission bracket 106 to fix the two shafts, the output double gear 101 and the small transmission gear 102 are respectively tightly mounted with the output transmission shaft 104 and the large transmission shaft 105, the driving stepping motor 116 and the differential stepping motor 117 are respectively tightly mounted with the motor connecting plate 115 and then tightly connected with the main body frame 107, the differential large gear 114 is tightly connected with the motor shaft of the differential stepping motor 117, the planet carrier 113 is in rolling connection with the motor shaft of the driving stepping motor 116 through the bearings, the driving pinion 111 is tightly connected with the motor shaft of the driving stepping motor 116, the driving pinion 111 is mounted above the planet carrier gear 111, the two double planet gears 112 are respectively tightly mounted with the two planet gear shafts 110, the two integral subassemblies are in rolling connection with the corresponding mounting holes on the planet carrier 113 through the bearings, the sun gear 109 and the large transmission shaft 108 pass through the central hole of the planet carrier 108, the planet carrier 108 is tightly mounted with the planet carrier 108, the planet carrier 84 is also in rolling connection with the planet carrier frame 108, and the planet carrier 84 is also has a rolling connection with the planet carrier 84, and a bearing mounted with the planet carrier 84.

In the actual movement shown in fig. 11, the differential stepping motor 117 is kept stationary in the torque mode, so that the carrier gear 113 is also stationary and the revolution of the two double planetary gears 112 is limited, and only the double planetary gears 112 are allowed to rotate for motion transmission, while the active stepping motor 116 is rotated, at this time, fixed transmission ratio fixed gear train reduction gear sets are provided, in which case, the torque mode of the differential stepping motor 117 is cancelled, and given its corresponding same rotation speed with respect to the active stepping motor 116, it will drive the differential gearwheel 14 and the carrier gear 113 engaged therewith to rotate, while the carrier gear 113 will drive the double planetary gear 112 to revolve around the sun gear 109, and since the rotation speeds of the two motors are the same, the revolution direction of the double planetary gear 112 is opposite to that of the active stepping motor 116, so that the rotation speed ratio calculated according to the transmission ratio of the differential planetary gear will be greatly changed and amplified by the original fixed shaft transmission ratio value, and the final rotation ratio of the stepping motor 117 will be changed if the rotation ratio of the two motors is the same or the ratio of the stepper motor 117 is changed.

The present invention is not limited to the above description of the embodiments, and those skilled in the art should, in light of the present disclosure, appreciate that many changes and modifications can be made without departing from the spirit and scope of the invention.

Claims

The microscopic motion tracking system is characterized by comprising an equipment base (1), a front vertical plate (2), a rear vertical plate (3), a sample fixing device (4) and a transverse plate (5), the equipment base (1), the front vertical plate (2), the rear vertical plate (3) and the transverse plate (5) are fixedly connected through bolts to form stable whole bodies, the sample fixing device (4) is fixed in an opening of the transverse plate (5) and keeps stable, a Y-direction moving base (8) is connected with the equipment base (1) through 2Y-direction linear guide rails (7) and a Y-direction driving motor and a ball screw (6), meanwhile, the X-direction moving base (14) is fixed on a through 2 triangular supports and a Y-direction moving base (8), an upper light path mounting support plate (10) and a lower light path mounting support plate (10) are connected with the X-direction moving base (14) through 2X-direction linear guide rails (16) and an X-direction driving motor, the ball screw (15) and the X-direction moving base (14), an upper light path system (13) and a lower light path mounting plate (10) are mounted above the upper light path mounting plate (10), and a lower light path mounting frame (11) and two micro-mounting devices (11) and a lower light path mounting rack (10) are mounted in front of the micro-operation system.
2. The micro-motion tracking system of claim 1, wherein the entire upper and lower optical systems and the micro-manipulator are connected to form bodies and can move in any direction in a plane under the drive of X, Y-direction motor.