CN113598709A

CN113598709A - Medicine screening device based on in vivo visible light imaging of living animal

Info

Publication number: CN113598709A
Application number: CN202110891436.4A
Authority: CN
Inventors: 刘松柏
Original assignee: Suzhou Vocational Health College
Current assignee: Suzhou Vocational Health College
Priority date: 2021-08-04
Filing date: 2021-08-04
Publication date: 2021-11-05

Abstract

The invention discloses a drug screening device based on in vivo visible light imaging of living animals, which comprises: the drug screening device comprises an equipment box, a camera bellows, an electron multiplication CCD arranged in the equipment box, an excitation light source, a PMT detector, a detection lens and a camera arranged in the camera bellows, and further comprises an animal fixing component, a first rotary driving mechanism, a temperature control component and a three-dimensional displacement mechanism, wherein the animal fixing component is rotatably arranged in the camera bellows, the first rotary driving mechanism is used for driving the animal fixing component to rotate around a Y axis, the temperature control component is used for providing wind heating for a living animal fixed on the animal fixing component, and the three-dimensional displacement mechanism is arranged in the camera bellows. According to the invention, the probe of the PMT detector can be automatically moved through the three-dimensional displacement mechanism, and the fixed living animal can be rotated through the first rotation driving mechanism and the animal fixing component, so that on one hand, the whole animal can be conveniently optically detected, on the other hand, the animal can be conveniently matched with the three-dimensional displacement mechanism for local detection, and the operation is simpler, more convenient and faster in the detection process.

Description

Medicine screening device based on in vivo visible light imaging of living animal

Technical Field

The invention relates to the technical field of drug screening, in particular to a drug screening device based on in-vivo visible light imaging of living animals.

Background

Currently, with the increasing severity of the drug resistance problem, the development of novel antibacterial drugs and antitumor drugs for treating diseases related to microbial infection and treating tumors has become an urgent problem to be solved.

Drug screening is an important step in modern drug development processes for selecting products with a certain potency from a large number of compounds or new compounds by means of standardized experiments. At present, the more commonly used drug screening process is a plurality of links such as animal infection or transplantation tumor inoculation, drug administration, animal sacrifice, organ grinding, viable bacteria counting and the like, and the method has a plurality of defects, such as: complicated operation, long drug development period and high cost. Drug screening by in vivo imaging of living animals can solve these problems.

The in vivo imaging technology for living animals refers to a technology for qualitatively and quantitatively researching tissue, cell and molecular levels of biological processes in a living state by applying an imaging method, and can be used for drug research and development. The in vivo visible light imaging (optical in vivo imaging) technology mainly includes two technologies of bioluminescence (bioluminescence) and fluorescence (fluorescence) imaging. In the bioluminescence imaging, luciferase (luciferase) gene is used for marking cells or DNA, and protease generated by the luciferase gene and a corresponding substrate are used for generating a probe light signal in an organism through a biochemical reaction; the fluorescence imaging is to label a fluorescent substance, form a fluorescent light source in a body by using a fluorescence labeling substance under the action of an exciting light source, collect optical signals by using a highly sensitive optical detection device, and evaluate the drug effect of the drug by using an optical detection device (such as a highly sensitive CCD (charge coupled device), a PMT (photomultiplier tube) weak light detector and the like) so as to realize drug screening. In the living body fluorescence imaging, labeling methods such as fluorescent protein labeling (fluorescent protein is suitable for labeling tumor cells, viruses, bacteria, genes and the like, and generally used are GFP, EGFP, RFP and the like), fluorescent dye labeling (commonly used are Cy3, Cy5, Cy5.5 and Cy7, and antibodies, polypeptides, small molecule drugs and the like can be labeled), quantum dot labeling and the like can be adopted, and a fluorescent substance is injected into an animal body, is irradiated by an excitation light source, and then fluorescence data transmitted out of the animal is collected, so that analysis such as drug targeting research and the like can be carried out, and the method can be used for drug screening. For example, some antibacterial drug screening systems can distinguish drug effects by detecting animal body luminescence signals, so as to realize drug screening, but because the exposure imaging requires a long time, animals need to be anesthetized during imaging detection, and the luminous intensity of the animals before and after anesthesia has a large influence, so that the measurement result has a large error; and these systems cannot stably measure the luminescence value of the whole living animal and accurately measure the local luminescence value of the animal.

In order to solve the above problems, patent CN109998480A discloses an in vivo drug screening system, which can perform drug screening by visible light detection, can meet the requirements of rapid measurement and accurate counting of local weak light in the animal body, and can perform overall observation and detection of the weak light in the animal body. When local precise measurement is carried out, the animal can be directly measured in vivo without anesthesia, the luminous value is stable, the result is accurate, and the measured data value can be directly used for objective evaluation of the medicine without other processing. The patent provides a good idea for the living animal drug screening technology, and has higher reference value for drug screening research, but the scheme still has at least the following defects:

1. when this scheme carries out animal overall observation and detects, fix the animal and place the constant temperature platform at the biology, the exciting light source shines from the top, and electron multiplication CCD's camera lens is also only followed the top and is gathered light signal for the whole body of animal can not obtain even, abundant exciting light irradiation, and electron multiplication CCD's camera lens also can not be even, the whole fluorescence that sends of abundant collection animal, makes optical detection's sensitivity and degree of depth receive the restriction, can cause great error to the result of whole detection.

2. In the scheme, when local luminescence measurement is carried out, an operator needs to hold a small animal through a light-proof glove and attach an infected part on a PMT weak light detection counting device for measurement; on one hand, the mode is troublesome to operate, and the small animals need to be firstly released and fixed from the biological placing constant temperature table and then caught to a PMT weak light detection counting device; on the other hand, the light-resistant gloves block easily that the partial local position that awaits measuring sends fluorescence, lead to measuring result to receive the influence, and snatch the live body toy and will await measuring the operation on the position attaches PMT weak light detection counting assembly has certain degree of difficulty, has increased the detection operation degree of difficulty, and because of the misoperation influences the result easily, has also increased operating personnel's work load simultaneously, when carrying out continuous operation, can cause great burden to operating personnel.

Therefore, there is a need for improvements in current drug screening systems to provide more reliable solutions.

Disclosure of Invention

The technical problem to be solved by the present invention is to provide a drug screening device based on in vivo visible light imaging of living animals, aiming at the above-mentioned deficiencies in the prior art.

In order to solve the technical problems, the invention adopts the technical scheme that: a drug screening device based on in vivo visible light imaging of a living animal, comprising: the drug screening device comprises an equipment box, a camera bellows, an electron multiplication CCD arranged in the equipment box, an excitation light source, a PMT detector, a detection lens and a camera arranged in the camera bellows, wherein the detection lens is connected with the electron multiplication CCD;

the field of view of the detection lens covers the whole living animal on the animal fixing component so as to carry out whole optical detection on the living animal;

the probe of the PMT detector is arranged at the driving end of the three-dimensional displacement mechanism, and the three-dimensional displacement mechanism drives the probe to move in XYZ three axes so as to perform local light detection on the living animal on the animal fixing component.

Preferably, the animal fixing component comprises a supporting rod connected to the bottom surface of the top plate of the dark box, a rotating seat rotatably connected to the inner wall of the side plate of the dark box, and an animal fixing frame with one end rotatably connected to the supporting rod and the other end in driving connection with the rotating seat, and the first rotary driving mechanism drives the rotating seat to rotate so as to drive the animal fixing frame to rotate;

the animal fixing frame comprises a main beam arranged along the X direction, a head fixing plate slidably arranged on the main beam, a tail fixing plate slidably arranged on the main beam, an auxiliary fixing beam connected to the main beam and arranged along the Y direction, a first joint fixedly connected to the first end of the main beam and a second joint fixedly connected to the second end of the main beam;

the bottom of the supporting rod is connected with an installation seat, and the first joint is rotatably arranged on the installation seat;

the rotary seat is provided with a driving jack for the second connector to be matched and inserted, and the second connector is in driving connection with the rotary seat through a key groove structure.

Preferably, the first rotary driving mechanism includes a first motor disposed on an outer wall of a side plate of the camera bellows, a driving gear drivingly connected to an output shaft of the first motor, and a driven gear fixedly sleeved on the rotary base and engaged with the driving gear.

Preferably, the mounting seat comprises a main mounting seat fixedly connected with the bottom of the supporting rod, a main mounting hole formed in the main mounting seat along the X direction, a sub mounting seat slidably arranged in the main mounting hole along the X direction, a sub mounting hole formed in the sub mounting seat along the X direction, and a pressure spring connected between the inner wall of the main mounting hole and the sub mounting seat;

the inner wall of the main mounting hole is also provided with a limiting guide groove along the X direction, and the outer wall of the sub mounting seat is connected with a limiting guide block which is slidably arranged in the limiting guide groove;

the first joint is rotatably arranged in the sub mounting hole.

Preferably, cavities are formed inside the main beam, the auxiliary fixing beam and the second joint, air outlet holes communicated with the cavities are formed in the surfaces of the main beam and the auxiliary fixing beam, and a horn mouth is formed at the tail end of the cavity of the second joint;

the side plate of the camera bellows is provided with an air supply outlet, the rotary seat is provided with an air supply channel communicated with the air supply outlet and the driving jack, the animal fixing frame is connected to the rear of the rotary seat, and the air supply outlet, the air supply channel, the horn mouth and the cavity are communicated in sequence.

Preferably, the temperature control assembly comprises a sealing cover connected to a side plate of the dark box, a fan arranged in the sealing cover, a heater arranged in the air supply port and a temperature sensor arranged in the air supply channel;

the first motor is arranged inside the sealing cover, an air inlet is formed in the sealing cover, the fan is arranged below the first motor, the air supply outlet is positioned below the first motor, and an arc-shaped air deflector is arranged below the inside of the sealing cover;

under the action of the fan, air outside the sealing cover enters the sealing cover through the air inlet, then is blown to the first motor, then enters the cavity after sequentially passing through the air supply outlet, the air supply channel and the horn mouth, and finally is discharged from the air outlet holes in the main beam and the auxiliary fixing beam and blown to the living animals fixed on the animal fixing frame.

Preferably, a reflecting cover is further arranged on the bottom plate of the dark box, the reflecting cover is positioned below the animal fixing frame, and a box door is arranged at the front part of the dark box;

the equipment box is also internally provided with a main control module and a power supply module, and a touch display screen is arranged on a front panel of the equipment box.

Preferably, a side plate of the dark box is provided with a ventilation port, the inner end of the ventilation port is provided with a filter, and the outer end of the ventilation port is provided with a light blocking assembly;

the light blocking assembly comprises a light blocking block connected to a side plate of the dark box, a ventilation channel arranged in the light blocking block and communicated with the ventilation port, and a light shield arranged on the light blocking block;

the air exchange channel comprises an inner port communicated with the air exchange port, a U-shaped channel communicated with the inner port and an outer port communicated with the U-shaped channel, and the inner port and the outer port are conical ports with large outer parts and small inner parts;

the light shield cover is arranged above the outer port, the light shield comprises a light shield cylinder connected with the light blocking block and a sealing cover plate connected to the light shield cylinder, a grid hole is formed in the light shield cylinder, and the ventilation channel is communicated with the external environment through the grid hole.

Preferably, the three-dimensional displacement mechanism comprises a Y-direction displacement mechanism arranged at the side part of the reflector, an X-direction displacement mechanism arranged on the Y-direction displacement mechanism, a Z-direction displacement mechanism arranged on the X-direction displacement mechanism, and a driving block arranged on the Z-direction displacement mechanism, wherein the driving block can move in three directions of XYZ under the driving of the Y-direction displacement mechanism, the X-direction displacement mechanism and the Z-direction displacement mechanism;

the driving block is also connected with a second rotary driving mechanism, the probe is arranged on the second rotary driving mechanism, and the rotating mechanism is used for driving the probe to rotate around an X axis;

the second rotary driving mechanism comprises a second motor arranged on the driving block and a rotary block in driving connection with an output shaft of the second motor, and the probe is connected to the rotary block.

Preferably, the rotating block comprises a main board body in driving connection with the output shaft of the second motor and a buffer board body connected with the back surface of the main board body, and the probe is connected to the buffer board body;

the mounting groove has been seted up at the back of the mainboard body, be provided with a plurality of guide bars in the mounting groove, be connected with the cooperation on the buffering plate body and insert and establish sliding connection piece in the mounting groove, the confession has been seted up on the sliding connection piece the guide bar cooperation male guiding hole, the upper portion and the lower part of guide bar are equipped with buffer spring and lower buffer spring respectively, it is in to go up buffer spring between the upper portion inner wall of mounting groove and the upper surface of sliding connection piece, lower buffer spring is in between the bottom inner wall of mounting groove and the lower surface of sliding connection piece, sliding connection piece can be relative the guide bar makes a round trip to slide.

The invention has the beneficial effects that:

1. the invention can realize the optical detection of the whole living animal through the electron multiplication CCD and the detection lens, carry out the overall optical signal analysis and realize the primary screening of the medicine; the PMT detector is matched with the three-dimensional displacement mechanism to perform optical detection on the local position of the living animal, perform optical signal analysis on a local interested part and provide more accurate experimental data for drug screening;

2. according to the invention, the fixed living animal can be rotated through the first rotary driving mechanism and the animal fixing component, so that the whole body of the living animal can be optically detected. By rotating the living animal, the exciting light can fully irradiate the whole body of the living animal from all directions of the outer periphery of the animal, and meanwhile, the detection lens can fully collect the fluorescence emitted by the whole living animal, so that the sensitivity and the depth of optical detection can be improved, and the whole optical detection result is more accurate;

3. according to the invention, the living animal is fixed through the animal fixing frame, and the whole animal fixing frame is directly taken down when the living animal is taken down, so that the operations of repeatedly unfastening, connecting and fixing ropes, binding bands and the like are not needed, the living animal is not needed to be anesthetized, and meanwhile, the living animal is fixed, and the injection of the medicine is more convenient;

4. according to the invention, the second rotary driving mechanism can drive the probe of the PMT detector to rotate around the X axis, and the position of the probe can be quickly and conveniently adjusted by matching with the first rotary driving mechanism and the three-dimensional displacement mechanism, so that the probe can move close to the local position of the living animal to perform optical detection; in some further embodiments, the buffer mechanism arranged in the rotating block can enable the probe to be tightly attached to the skin of the living animal without exerting excessive force on the probe, and by exerting flexible acting force, the probe can be prevented from moving excessively to crush the animal or damage the probe, and meanwhile, the probe can be ensured to be tightly attached to the skin of the living animal as much as possible, so that the reliability of the detection result is improved;

5. according to the invention, the temperature control assembly provides air heating, so that an even and mild heat source can be provided for living animals, a mild constant temperature environment close to the body temperature is provided, and the influence of temperature factors on detection results can be reduced; the temperature of the hot air can be controlled to be maintained in a set range through the feedback of the temperature sensor, so that the live animals are prevented from being scalded, the rotating function of the animal fixing frame can be adapted, and the problem of line winding can be avoided; in some further embodiments, the heat dissipation problem of the first motor can be solved through the action of the fan in the temperature control assembly, the equipment is simplified, the heat dissipated by the first motor can be recovered, and the energy consumption can be reduced.

Drawings

Fig. 1 is a schematic diagram of the internal structure of a drug screening device based on visible light imaging in vivo of a living animal in one embodiment of the present invention;

fig. 2 is a front view of a drug screening device based on visible light imaging in vivo of a living animal in an embodiment of the present invention;

FIG. 3 is a schematic top view of a three-dimensional displacement mechanism according to an embodiment of the present invention;

FIG. 4 is a schematic view of an animal mount according to an embodiment of the present invention;

fig. 5 is a schematic structural view of an animal holder cooperating with a first rotary driving mechanism and a temperature control assembly according to an embodiment of the present invention;

FIG. 6 is a partial block diagram of FIG. 5 of the present invention;

FIG. 7 is a schematic cross-sectional view of an animal mount in one embodiment of the invention;

FIG. 8 is a schematic view of a mounting base and a first connector according to an embodiment of the present invention;

FIG. 9 is a schematic structural view of a main mount in an embodiment of the present invention;

FIG. 10 is a cross-sectional structural schematic view of a main mount in cross-section in one embodiment of the invention;

FIG. 11 is a schematic structural view of a sealing boot in an embodiment of the present invention;

FIG. 12 is a schematic structural diagram of a light barrier assembly in an embodiment of the present invention;

FIG. 13 is a schematic view of a light shield disposed on a sealing cap according to an embodiment of the present invention;

FIG. 14 is a schematic structural view of a three-dimensional displacement mechanism in an embodiment of the present invention;

FIG. 15 is a schematic view of another perspective of a three-dimensional displacement mechanism in an embodiment of the present invention;

FIG. 16 is a schematic view of a second rotary drive mechanism coupled to a probe in accordance with an embodiment of the present invention;

FIG. 17 is a schematic view of an X-direction displacement mechanism in an embodiment of the present invention;

FIG. 18 is a schematic longitudinal sectional view of a rotating block in an embodiment of the present invention;

fig. 19 is a cross-sectional structural view of a rotation block in an embodiment of the present invention.

Description of reference numerals:

1-equipment box; 10-electron multiplying CCD; 11-a master control module; 12-a power supply module; 13-a touch display screen; 14-light source control module; 15-power connection;

2-dark box; 20-side plate; 21-top plate; 22-a base plate; 23-a box door; 24-excitation light source; 25-PMT detector; 26-detection lens; 27-a camera; 28-a reflector; 200-air supply outlet; 201-a ventilation port; 202-a filter; 230-door lock; 250-a probe; 251-PMT detector body;

3-an animal fixation assembly; 30-a support bar; 31-a rotating base; 32-an animal mount; 33-a mounting seat; 34-locking screws; 310-a drive jack; 311, a transmission groove; 312-a bearing; 313-flange plate; 314 — air supply channel; 320-main beam; 321-head fixing plate; 322-tail fixing plate; 323-auxiliary fixed beam; 324 — a first joint; 325 — second joint; 326 — a cavity; 327-air outlet; 328-a fixed rope; 329-band; 330-main mounting seat; 331 — main mounting hole; 332-sub mount; 333-sub mounting hole; 334-compression spring; 335-limit guide groove; 336-limit guide block; 337-bearings; 3210-a tab; 3250 — drive key; 3260-horn mouth;

4-a first rotary drive mechanism; 40-a first motor; 41-a transmission shaft; 42-a drive gear; 43-driven gear;

5-temperature control component; 50-a sealing cover; 51-a fan; 52-a heater; 53-temperature sensor; 500-air inlet; 501-arc wind deflector; 502-door of the bin; 503-sealing door;

6-a three-dimensional displacement mechanism; 60-Y direction displacement mechanism; 61-X direction displacement mechanism; 62-Z displacement mechanism; 63-a drive block; 64-a second rotary drive mechanism; 610-X motor; 611-X lead screw; 612-X slide rail; 613-X slider; 640 — a second electric machine; 641 — rotation block; 642-main board body; 643 — a buffer plate body; 644-reducer; 6420-mounting groove; 6421-guide bar; 6422-upper buffer spring; 6423-lower buffer spring; 6430-sliding connection block; 6431-pilot hole; 6432-sliding sleeve;

7-a light barrier assembly; 70-light blocking block; 71-a ventilation channel; 72-a light shield; 710 — an inner port; 711-U-shaped channel; 712 — an external port; 713-horizontal channel; 720, a shading cylinder; 721, sealing a cover plate; 722-grid holes.

Detailed Description

The present invention is further described in detail below with reference to examples so that those skilled in the art can practice the invention with reference to the description.

It will be understood that terms such as "having," "including," and "comprising," as used herein, do not preclude the presence or addition of one or more other elements or groups thereof.

Example 1

As shown in fig. 1 to 3, a drug screening apparatus based on visible light imaging in vivo of a living animal according to the present embodiment includes: the drug screening device comprises an equipment box 1, a camera box 2, an electron multiplying CCD10 arranged in the equipment box 1, an excitation light source 24, a PMT detector 25, a detection lens 26 and a camera 27 arranged in the camera box 2, wherein the detection lens 26 is connected with the electron multiplying CCD 10;

the visual field of the detection lens 26 covers the whole live animal on the animal fixing component 3 so as to carry out whole optical detection on the live animal;

the probe 250 of the PMT detector 25 is disposed at the driving end of the three-dimensional displacement mechanism 6, and the three-dimensional displacement mechanism 6 drives the probe 250 to perform XYZ three-axis movement so as to perform local optical detection on the living animal on the animal fixing assembly 3.

The device of the invention is based on the in vivo fluorescence imaging of the living animal to screen the medicine, firstly, the fluorescence labeling is carried out by means of fluorescent protein or fluorescent dye, the fluorescence can be generated on the living animal body when the excitation light source 24 irradiates, and the action effect of the medicine can be researched by the methods of detecting, counting, imaging and the like on the fluorescence, thereby realizing the medicine screening. In addition, the invention can realize the optical detection of the whole living animal through the electron multiplication CCD10 and the detection lens 26, and carry out the global optical signal analysis; the invention can realize the automatic movement of the probe 250 of the PMT detector 25 through the three-dimensional displacement mechanism 6, and can rotate the fixed living animal through the first rotary driving mechanism 4 and the animal fixing component 3, thereby being convenient for realizing the optical detection of the whole animal on one hand, and being convenient for matching with the three-dimensional displacement mechanism 6 to carry out the local detection on the other hand, and the operation is simpler, more convenient and faster in the detection process. Furthermore, in the detection process, the temperature control assembly 5 provides air heating, so that a mild constant temperature environment close to the body temperature of the small animal is provided for the small animal, and the influence of temperature factors on the detection result can be reduced.

The device according to the present invention may be used in a variety of ways to perform drug screening studies, for example, in one embodiment, the overall process of drug screening using the device is:

1. firstly, carrying out fluorescence labeling on a target strain acted by a drug to be selected, then injecting the strain into a small animal body, fixing the small animal on an animal fixing component 3 after a certain time, carrying out integral optical detection through an electron multiplication CCD10 and a detection lens 26, and initially counting fluorescence photons (fluorescence intensity); then injecting the drug to be selected, carrying out integral optical detection again after a certain time, and comparing the difference of the number of fluorescence photons (the difference of fluorescence intensity) of two times, thereby preliminarily judging the effect of the drug to be selected and rapidly screening the drug with certain drug effect from a large amount of developed drugs;

2. the method can further and accurately screen the medicines with certain drug effects through local detection, and comprises the following steps: injecting the target strain after fluorescent marking into the small animal body, fixing the small animal on the animal fixing component 3 after a certain time, observing the main action area of the medicine as a local interested position through integral optical detection, and then carrying out optical detection on the local interested position on the small animal through a PMT detector 25; and then injecting the medicine with certain efficacy into the small animal, performing local optical detection again after a certain time, comparing the results of the two times, further determining the efficacy and the main action part of the medicine, and providing information for the screening of the medicine, thereby realizing efficient and accurate medicine screening.

Of course, the device of the present invention may also be used for drug screening studies using other procedures based on its structure.

The electron multiplying CCD10(EMCCD) can realize the rough measurement and detection of the weak light intensity of the whole living animal; the PMT detector 25 is a PMT weak light detector, is tightly attached to a living animal through the probe 250, and can realize accurate measurement and detection of local weak light intensity.

In a preferred embodiment, the bottom plate 22 of the dark box 2 is further provided with a reflector 28, the reflector 28 is located below the animal fixing frame 32, on one hand, the excitation light source 24 can be sufficiently irradiated to each part of the living animal through the reflector 28, on the other hand, fluorescence emitted by the living animal can be sufficiently collected by the detection lens 26, and the accuracy of the detection result is improved.

In a preferred embodiment, a door 23 is provided at the front of the camera chamber 2, a door lock 230 is further provided on the door 23, and a sealing strip is provided between the door 23 and the camera chamber 2 to ensure that no light enters the camera chamber 2 after the door 23 is closed.

In a preferred embodiment, a main control module 11, a power supply module 12 and a light source control module 14 are further disposed in the equipment box 1, and a touch display screen 13 is disposed on a front panel of the equipment box 1. The electron multiplying CCD10, the PMT detector 25, the touch display screen 13, the light source control module 14, the camera 27 and the like are connected with the main control module 11. The excitation light source 24 is connected to the light source control module 14, and the light source control module 14 is configured to control the excitation light source 24 to emit a desired laser light source. The main control module 11 may adopt a microcomputer or a control chip, and can implement a control function of each component and a data processing function. The touch screen 13 can display the results of the detection and can provide an input interface for controlling the device and setting the relevant parameters. The power module 12 is used to provide power. In a further embodiment, a power connector 15 is included for connection to an external power source.

The camera 27 is preferably an infrared camera, which can image a living animal, can display an image by touching the display screen 13, and can also guide the operation and control of the three-dimensional displacement mechanism 6 inside the dark box 2.

The foregoing is a general idea of the present invention, and more specific embodiments thereof are provided below.

Example 2

Referring to fig. 4 to 10, the animal fixing member 3 is used for fixing a small living animal for optical detection, and can realize rotation of the small living animal, and can be conveniently detached and removed to meet the operation requirement in the detection process.

As a further improvement on the basis of embodiment 1, in this embodiment, the animal fixing assembly 3 includes a supporting rod 30 connected to the bottom surface of the top plate 21 of the dark box 2, a rotating base 31 rotatably connected to the inner wall of the side plate 20 of the dark box 2, and an animal fixing frame 32 having one end rotatably connected to the supporting rod 30 and the other end drivingly connected to the rotating base 31, and the first rotary driving mechanism 4 drives the rotating base 31 to rotate to drive the animal fixing frame 32 to rotate; in this embodiment, a flange 313 is fixed on the inner wall of the side plate 20 of the dark box 2, and the rotary base 31 is rotatably connected to the flange 313 through a bearing 312.

The animal holder 32 includes a main beam 320 disposed along the X-direction, a head fixing plate 321 slidably disposed on the main beam 320, a tail fixing plate 322 slidably disposed on the main beam 320, an auxiliary fixing beam 323 attached to the main beam 320 and disposed along the Y-direction, a first joint 324 fixed to a first end of the main beam 320, and a second joint 325 fixed to a second end of the main beam 320.

The tail end of the head fixing plate 321 is provided with a protruding piece 3210 for placing the head of the small animal, the head fixing plate 321 and the tail fixing plate 322 are both connected with a fixing rope 328 or other fixing structures (such as buckles) for fixing the limbs of the small animal, the two ends of the auxiliary fixing beam 323 are also connected with a binding belt 329 or other fixing structures (such as buckles) for fixing the body of the small animal, and the small animal can be firmly fixed on the animal fixing frame 32 through the fixing rope 328 and the binding belt 329.

In an embodiment, the main beam 320, the auxiliary fixing beam 323, the head fixing plate 321, and the tail fixing plate 322 are made of transparent materials (such as acrylic materials), so that light can pass through the main beam, and the influence on optical detection is reduced.

Rectangular sliding holes for the main beams 320 to pass through are formed in the middle of the head fixing plate 321 and the tail fixing plate 322, locking screws 34 are connected to the head fixing plate 321 and the tail fixing plate 322 in a threaded mode, the inner ends of the locking screws 34 extend into the rectangular sliding holes, and the head fixing plate 321 and the tail fixing plate 322 can be fixed to the main beams 320 by jacking the outer walls of the main beams 320. When the device is used, the positions of the head fixing plate 321 and the tail fixing plate 322 on the main beam 320 are adjusted in a sliding mode according to the size of a small living animal, and the locking screws 34 are screwed after the adjustment is completed, so that the head fixing plate 321 and the tail fixing plate 322 are kept fixed. The rectangular main beams 320 and the rectangular sliding holes are matched, so that the head fixing plate 321 and the tail fixing plate 322 can slide along the X direction but cannot rotate.

The bottom of the support rod 30 is connected with a mounting seat 33, and a first joint 324 is rotatably arranged on the mounting seat 33;

the rotary seat 31 is provided with a driving jack 310 for the second connector 325 to be inserted in a matching manner, and the second connector 325 is in driving connection with the rotary seat 31 through a key groove structure. In this embodiment, the second connector 325 is provided with a transmission key 3250, the inner wall of the driving jack 310 is provided with a transmission groove 311, and the transmission key 3250 is inserted into the transmission groove 311 in a matching manner, so that the second connector 325 can be driven to rotate by the rotating base 31.

The first rotary driving mechanism 4 includes a first motor 40 disposed on the outer wall of the side plate 20 of the dark box 2, a transmission shaft 41 drivingly connected to an output shaft of the first motor 40, a driving gear 42 connected to the transmission shaft 41, and a driven gear 43 fixedly sleeved on the rotary base 31 and engaged with the driving gear 42. The first motor 40 drives the driving gear 42 to rotate, and then the rotary seat 31 is driven to rotate by the driven gear 43, and the rotary seat 31 drives the animal fixing frame 32 and the living animal thereon to rotate. In a preferred embodiment, the first motor 40 is selected from a stepper motor, and the speed reducer 644 can be disposed between the first motor 40 and the driving gear 42, or the speed reduction can be achieved by configuring the driving gear 42 to have a smaller outer diameter than the driven gear 43. Wherein, set up the shaft hole that supplies transmission shaft 41 to pass on the curb plate 20 of camera bellows 2, need make sealedly between shaft hole and the transmission shaft 41, prevent that light from getting into inside camera bellows 2.

The mounting seat 33 comprises a main mounting seat 330 fixedly connected with the bottom of the support rod 30, a main mounting hole 331 formed in the main mounting seat 330 along the X direction, a sub mounting seat 332 slidably arranged in the main mounting hole 331 along the X direction, a sub mounting hole 333 formed in the sub mounting seat 332 along the X direction, and a pressure spring 334 connected between the inner wall of the main mounting hole 331 and the sub mounting seat 332; the inner wall of the main mounting hole 331 is also provided with a limiting guide groove 335 along the X direction, and the outer wall of the sub-mounting seat 332 is connected with a limiting guide block 336 which is slidably arranged in the limiting guide groove 335; the first joint 324 is rotatably disposed in the sub-mounting hole 333. In one embodiment, the first connector 324 is rotatably disposed within the sub-mounting hole 333 via a bearing 337.

The animal holder 32 is detachably provided on the mount 33 and the rotary base 31, and is rotated by the driving of the first rotary drive mechanism 4, thereby enabling the living animal to perform optical detection of the whole body. By rotating the living animal, the exciting light can fully irradiate the whole body of the living animal from all directions of the outer periphery of the animal, and meanwhile, the detection lens 26 can fully collect the fluorescence emitted by the whole living animal, so that the sensitivity and the depth of optical detection can be improved, and the whole optical detection result is more accurate. For example, in one embodiment, the average fluorescence intensity (e.g., optical signal such as fluorescence photon amount) detected by the detection lens 26 during one or more cycles of the rotation of the living animal is used as the overall optical detection result, so that the result has less error and is more valuable for drug screening and evaluation. On the other hand, by rotating the living animal, the local position of the living animal can be detected by the PMT detector 25 conveniently without grasping the living animal by hand (further described in the following embodiments).

The principle of the detachable installation of the animal fixing frame 32 on the installation seat 33 and the rotating seat 31 is as follows: when in use, the animal fixing frame 32 is taken down, and the small living animal is fixed on the animal fixing frame 32;

referring to fig. 8, the sub-mount 332 is pressed leftward to retract the sub-mount 332 into the main mounting hole 331, and the first connector 324 of the animal holder 32 is inserted into the sub-mounting hole 333 of the sub-mount 332;

referring to fig. 5 and 6, the animal holder 32 is rotated to align the driving key 3250 on the second connector 325 with the driving groove 311 on the rotary base 31, and the second connector 325 is aligned with the driving insertion hole 310 (the second connector 325 is aligned with the driving insertion hole 310), then the sub-mount 332 is released, the sub-mount 332 extends out of the main mounting hole 331 under the elastic force of the compression spring 334, the first connector 324 of the animal holder 32 is pressed to move the animal holder 32 to the right, the second connector 325 is inserted into the driving insertion hole 310 to be mounted in place, the animal holder 32 is stably arranged on the mount 33 and the rotary base 31, and when the rotary base 31 rotates the animal holder 32, the first connector 324 can freely rotate in the sub-mounting hole 333.

When the animal fixing frame 32 is taken down, the sub-mounting seat 332 is pressed leftwards, the sub-mounting seat 332 is retracted into the main mounting hole 331, then the first joint 324 of the animal fixing frame 32 is moved, the first joint 324 is separated from the sub-mounting hole 333, and then the second joint 325 is pulled out of the rotating seat 31, so that the animal fixing frame 32 can be conveniently taken down.

In the detection process, the living animal is usually required to be taken down for operation for a plurality of times, for example, after the whole optical detection is carried out on the living animal after the bacterial strain is injected, the living animal is taken out for injecting the medicine, then the detection is carried out, and then the local optical detection is carried out and the medicine injection is carried out on the living animal; if the living animal is directly taken off from the animal fixing frame 32, the operation is troublesome (a plurality of fixing ropes, binding bands and the like need to be uncovered, the fixing ropes, the binding bands need to be connected again when the living animal is re-fixed, and the living animal needs to be anesthetized in some cases). In this embodiment, the animal holder 32 is directly removed as a whole, the living animal is always fixed to the animal holder 32, the operations of repeatedly unfastening and connecting the fixing string 328 and the strap 329 are not required, anesthesia is not required for the living animal, and the injection of the medicine is more convenient because the living animal is fixed.

Example 3

In the detection process, a constant temperature environment close to the body temperature of the living small animal needs to be provided for the living small animal so as to maintain the normal physiological state of the living small animal and reduce the influence of temperature factors on the measurement result. In the prior art, a living small animal is usually fixed on a constant temperature platform and is heated by contacting a heating plate or a heating sheet and the like with the living small animal, so that the defects of uneven heating, non-mild heating, easy scald on the small animal and the like exist; in addition, the mechanism for fixing the small animal in the invention also needs to provide a rotating function, and the problem that the line is wound due to rotation is easy to occur by adopting the scheme in the prior art, so the constant temperature control scheme in the prior art is difficult to meet the requirement. The present invention provides a constant temperature environment by the structure of the following embodiments.

Referring to fig. 5 and 7, as a further improvement on the basis of embodiment 2, in this embodiment, cavities 326 are provided inside the main beam 320, the auxiliary fixing beam 323, and the second joint 325, a plurality of air outlets 327 communicating with the cavities 326 are densely provided on the surfaces of the main beam 320 and the auxiliary fixing beam 323, and a bell mouth 3260 is provided at the end of the cavity 326 of the second joint 325; the side plate 20 of the camera bellows 2 is provided with an air supply outlet 200, the rotary seat 31 is provided with an air supply channel 314 communicated with the air supply outlet 200 and the driving jack 310, and after the animal fixing frame 32 is connected to the rotary seat 31, the air supply outlet 200, the air supply channel 314, the bell mouth 3260 and the cavity 326 are communicated in sequence. The flare 3260 can facilitate the entry of heated air in the supply air path 314 into the cavity 326.

Wherein, the temperature control assembly 5 comprises a sealing cover 50 connected with the side plate 20 of the dark box 2, a fan 51 arranged in the sealing cover 50, a heater 52 arranged in the air supply outlet 200 and a temperature sensor 53 arranged in the air supply channel 314;

the first motor 40 is arranged inside the sealing cover 50, the sealing cover 50 is provided with an air inlet 500, the fan 51 is arranged below the first motor 40, the air supply outlet 200 is arranged below the first motor 40, and an arc-shaped air deflector 501 is arranged below the inside of the sealing cover 50; the arc-shaped air deflector 501 plays a role in guiding the hot air passing through the first motor 40 to enter the air supply outlet 200.

Under the action of the fan 51, air outside the sealed enclosure 50 enters the sealed enclosure 50 through the air inlet 500, blows towards the first motor 40, then sequentially passes through the air supply outlet 200, the air supply channel 314 and the bell mouth 3260, enters the cavity 326, is finally discharged from the main beam 320 and the air outlet 327 on the auxiliary fixing beam 323, and blows towards the living animal fixed on the animal fixing frame 32.

In one embodiment, the fan 51, the heater 52 and the temperature sensor 53 are all connected to the main control module 11, and the temperature is controlled by the main control module 11. In another embodiment, the temperature control assembly 5 further comprises a control chip, and the fan 51, the heater 52 and the temperature sensor 53 are all connected with the control chip, and the temperature control is performed by the control chip.

The working principle of the temperature control component 5 is as follows: under the action of the fan 51, air outside the sealed enclosure 50 enters the sealed enclosure 50 through the air inlet 500, is blown to the first motor 40, takes away heat of the first motor 40, then enters the air supply opening 200 under the guide action of the arc-shaped air deflector 501, is heated to a set temperature by the heater 52 in the air supply opening 200, then sequentially enters the air supply channel 314 and the bell mouth 3260, enters the cavity 326, finally is discharged from the main beam 320, the auxiliary fixing beam 323 and the air outlet 327 on the second connector 325, is blown to the living animal fixed on the animal fixing frame 32, and provides a temperature environment close to the body temperature of the living animal through uniform and mild hot air. In the process, when the animal fixing frame 32 rotates, the hot air in the air supply channel 314 continuously enters the cavity 326, and the supply of the hot air is not affected. The temperature sensor 53 detects the temperature of the hot air in the air supply channel 314 in real time, and when the temperature of the hot air is higher than a set value, the main control module 11 or the control chip controls the heater 52 to reduce power or stop working, so as to ensure that the temperature is maintained within a set range.

In the embodiment, a uniform and mild heat source can be provided for the living animal through hot air, and the temperature of the hot air can be controlled to be maintained in a set range through the feedback of the temperature sensor 53, so that the living animal is prevented from being scalded, meanwhile, the rotating function of the animal fixing frame 32 can be adapted, and the problem of line winding cannot occur; on the other hand, the first motor 40 generates heat after working for a long time, the fan 51 blows air to the first motor 40, the heat dissipation problem of the first motor 40 can be solved, the equipment is simplified, heat dissipated by the first motor 40 can be recovered, and energy consumption can be reduced. It should be understood that, in general, the heat dissipation amount of the first motor 40 is limited, and the hot air generated by heat dissipation of the first motor 40 cannot reach the set temperature range, so that the heater 52 is required to assist heating.

Referring to fig. 11, in another alternative embodiment, a door 502 is further disposed below the sealing cover 50, an air outlet is further formed on the sealing cover 50, and a sealing door 503 is also disposed on the air outlet. In general, the door 502 is opened, the sealing door 503 is closed, and the hot air having passed through the first motor 40 enters the air blowing port 200. The embodiment can also meet the requirements of the following special cases: the environment temperature of the device is high, and hot air does not need to be introduced to provide a temperature environment; at this time, the door 502 is closed, the sealing door 503 is opened, and the hot air passing through the first motor 40 is directly discharged from the outlet and does not enter the air supply opening 200.

Example 4

Referring to fig. 1, 12 and 13, as a further improvement on the basis of the above embodiment 3, a ventilation opening 201 is provided on the side plate 20 of the dark box 2, a filter 202 is provided at an inner end of the ventilation opening 201, and a light blocking assembly 7 is provided at an outer end of the ventilation opening 201;

the light blocking assembly 7 comprises a light blocking block 70 connected to the side plate 20 of the dark box 2, a ventilation channel 71 arranged in the light blocking block 70 and communicated with the ventilation port 201, and a light shield 72 arranged on the light blocking block 70;

the ventilation channel 71 comprises an inner port 710 communicated with the ventilation port 201, a U-shaped channel 711 communicated with the inner port 710 and an outer port 712 communicated with the U-shaped channel 711, wherein the inner port 710 and the outer port 712 are tapered ports with large outer parts and small inner parts; the inner port 710 and the outer port 712 are communicated with the upper end of the U-shaped channel 711 through the horizontal channel 713, the two horizontal channels 713 are communicated through the U-shaped bent channel, air can smoothly circulate through the bent channel structure, and external light cannot enter the dark box 2.

In the preferred embodiment, the inner wall of the ventilation channel 71 has a black surface, and further preferably, a black light-absorbing surface (which may be made of a conventional light-absorbing material) is used to reduce the reflection of light, and further prevent the light from entering the interior of the dark box 2.

The light shielding cover 72 is disposed above the outer port 712, the light shielding cover 72 includes a light shielding cylinder 720 connected to the light blocking block 70 and a sealing cover 721 connected to the light shielding cylinder 720, the light shielding cylinder 720 is provided with a grid hole 722, and the ventilation channel 71 is communicated with the external environment through the grid hole 722.

In a preferred embodiment, the light-shielding cylinder 720 has black light-absorbing inner and outer surfaces to reduce light entering the grid holes 722, and air circulates outside the interior of the light-shielding cylinder 720 through the grid holes 722. The inner surface of the sealing cover 721 and the right end surface of the light blocking block 70 are also black light absorbing surfaces to reduce light reflection.

An air vent 201 is arranged on the side plate 20 of the dark box 2 to maintain the air requirement of the living animal during the detection process, and a filter 202 is arranged on the air vent 201 to filter the air entering and exiting. In alternative embodiments, active ventilation may be provided by a fan 51 or the like.

The interior of the camera bellows 2 needs to be kept in a dark state during detection, so that external light is prevented from entering, and the door 23 of the camera bellows 2 can be protected from light by arranging a sealing strip; at the scavenge port 201, realize through setting up the subassembly 7 that is in the light in the outside of curb plate 20, can satisfy the demand of taking a breath, can prevent again that light from passing through scavenge port 201 and getting into inside camera bellows 2.

The outside air enters the light shielding cylinder 720 through the grill holes 722, then enters the interior of the dark box 2 after sequentially passing through the outer port 712, the ventilation channel 71, the inner port 710, the ventilation port 201 and the filter 202, and the air in the dark box 2 can be discharged to the outside, so that the inside and outside ventilation of the dark box 2 is realized.

A small portion of the light irradiated onto the light shield 72 enters the light shielding cylinder 720 through the grid holes 722, the small portion of the light is substantially absorbed by the inner surface of the sealing cover 721 and the black light absorbing surface of the right end face of the light blocking block 70, and the remaining light is substantially completely absorbed by the inner surface of the right side of the ventilation channel 71 after entering the outer port 712, so that substantially no light enters the interior of the dark box 2 through the ventilation port 201, and the optical detection of the interior of the dark box 2 is not affected.

In the preferred embodiment, the air inlet 500 above the sealing cap 50 is also provided with a light shield 72, which can reduce the light entering the sealing cap 50; in a further preferred embodiment, the inner walls of the air supply opening 200, the air supply passage 314, the bell mouth 3260 and the cavity 326 of the second joint 325 are all black light absorbing surfaces, and since the air supply opening 200 is perpendicular to the air inlet 500, substantially no light can enter the interior of the dark box 2 through the air supply opening 200, the air supply passage 314, the bell mouth 3260 and the cavity 326, and the optical detection of the interior of the dark box 2 cannot be affected.

Example 5

Referring to fig. 14 to 19, as a further improvement of any one of embodiments 2 to 4, in this embodiment, the three-dimensional displacement mechanism 6 includes a Y-direction displacement mechanism 60 provided at a side portion of the reflector 28, an X-direction displacement mechanism 61 provided at the Y-direction displacement mechanism 60, a Z-direction displacement mechanism 62 provided at the X-direction displacement mechanism 61, and a drive block 63 provided at the Z-direction displacement mechanism 62, and the drive block 63 is capable of being driven by the Y-direction displacement mechanism 60, the X-direction displacement mechanism 61, and the Z-direction displacement mechanism 62 to perform XYZ three-directional movements. The three-dimensional displacement mechanism 6 needs to be arranged to ensure that it is not affected by the reflector 28.

The X-direction displacement mechanism 61, the Y-direction displacement mechanism 60 and the Z-direction displacement mechanism 62 all adopt conventional linear displacement mechanisms, such as a screw motor mechanism, a belt pulley mechanism or a push rod mechanism, and the three-dimensional driving function is realized through the combination of the three linear displacement mechanisms. For example, taking the X-direction displacement mechanism 61 as an example, in an embodiment, the X-direction displacement mechanism includes an X motor 610, an X lead screw 611 connected to the X motor 610 in a driving manner, an X slide rail 612, and an X slider 613 fitted over the X lead screw 611 and fitted with the X slide rail 612, and the displacement driving in the X direction is realized through the lead screw motor mechanism. The same mechanism can be used for the Y-displacement mechanism 60 and the Z-displacement mechanism 62, and the details thereof are omitted.

The driving block 63 is also connected with a second rotary driving mechanism 64, the probe 250 of the PMT detector 25 is arranged on the second rotary driving mechanism 64, and the rotary mechanism is used for driving the probe 250 to rotate around the X axis; the second rotary drive mechanism 64 includes a second motor 640 provided on the drive block 63 and a rotary block 641 drivingly connected to an output shaft of the second motor 640, and the probe 250 is connected to the rotary block 641. In other embodiments, the PMT detector 25 may be integrally mounted on the rotating block 641, but since this would increase the load on the second rotating driving mechanism 64, in a preferred embodiment, the PMT detector body 251 is mounted on the dark box 2 or the equipment box 1, only the probe 250 is mounted on the rotating block 641, the probe 250 is connected to the PMT detector body 251, and the PMT detector body 251 is connected to the main control module 11. The probe 250 detects the optical signal and transmits the optical signal to the PMT detector body 251, or the probe 250 detects the optical signal, converts the optical signal into an electrical signal and transmits the electrical signal to the PMT detector body 251 for further data processing, and the processed data is transmitted to the main control module 11.

Wherein, the top of the probe 250 is spherical, which is convenient for being closely contacted with the surface of the animal to detect the weak light.

In a preferred embodiment, a reducer 644 is drivingly connected to the output shaft of the second motor 640, and the output shaft of the reducer 644 is connected to the rotating block 641. The second motor 640 rotates the rotating block 641, so that the probe 250 of the PMT detector 25 can rotate around the X-axis, thereby facilitating adjustment of the position of the probe 250, and facilitating optical detection by moving the probe 250 proximate to the local position of the living animal. For example, in one embodiment, when local light detection is performed, the animal holder 32 to which the living animal is fixed is rotated to a proper angle by the first rotary drive mechanism 4, then the probe 250 is moved to a position near the target region to be detected of the living animal by the three-dimensional displacement mechanism 6, the angle of rotation of the probe 250 about the X axis is adjusted by the second rotary drive mechanism 64, and the probe 250 is raised by the Z-direction displacement mechanism 62 so that the probe 250 is closely attached to the target region to be detected, and local light detection is performed. Through the cooperation of the second rotary driving mechanism 64, the first rotary driving mechanism 4 and the three-dimensional displacement mechanism 6, the probe 250 can be conveniently and accurately moved to a target area to be detected.

Referring to fig. 18-19, in a further preferred embodiment, the rotation block 641 includes a main plate body 642 drivingly connected to the output shaft of the second motor 640, and a buffer plate body 643 connected to a rear surface of the main plate body 642, the probe 250 being connected to the buffer plate body 643; the back of the main plate body 642 is provided with an installation groove 6420, a plurality of guide rods 6421 are arranged in the installation groove 6420, the buffer plate body 643 is connected with a sliding connection block 6430 which is inserted into the installation groove 6420 in a matching manner, the sliding connection block 6430 is provided with a guide hole 6431 for the guide rod 6421 to be inserted in a matching manner, the upper portion and the lower portion of the guide rod 6421 are respectively sleeved with an upper buffer spring 6422 and a lower buffer spring 6423, the upper buffer spring 6422 is positioned between the inner wall of the upper portion of the installation groove 6420 and the upper surface of the sliding connection block 6430, the lower buffer spring 6423 is positioned between the inner wall of the bottom of the installation groove 6420 and the lower surface of the sliding connection block 6430, and the sliding connection block 6430 can slide back and forth relative to the guide rod 6421. In a preferred embodiment, a sliding sleeve 6432 is provided in the guide hole 6431, and the guide bar 6421 is fittingly inserted in the sliding sleeve 6432.

When local light detection is carried out, in order to improve the accuracy of a detection result, the probe 250 needs to be attached to the skin of a living animal as closely as possible, and when the probe 250 is driven to automatically move by a mechanical driving mechanism, if the operation is improper, the risk of excessively moving and injuring the animal or damaging the probe 250 exists; if the probe 250 is not tightly attached to the skin of the living animal, the accuracy of the detection result may be affected. In this embodiment, the above-mentioned buffer mechanism can overcome this problem, and the reliability of the device can be further improved. The specific principle is as follows: taking the probe 250 approaching the living animal from the lower direction as an example, when the rotating block 641 drives the probe 250 to move upwards, after the probe 250 contacts the skin of the animal, the rotating block 641 still moves upwards for a short distance, at this time, the main plate body 642 continues to move upwards, the buffer plate body 643 receives resistance due to the contact of the front end of the probe 250 with the living animal, at this time, the lower buffer spring 6423 is compressed, the upper buffer spring 6422 is stretched, and the main plate body 642 applies an upward acting force to the buffer plate body 643 through the two buffer springs, so that the probe 250 can be tightly attached to the skin of the living animal, no excessive force is applied to the probe 250, the probe 250 can be prevented from being excessively moved to crush the animal or damage the probe 250 by applying a flexible acting force, the probe 250 can be ensured to be tightly attached to the skin of the living animal as much as possible, and the reliability of the detection result is improved.

Example 6

This embodiment provides the drug screening method of the drug screening apparatus based on in vivo visible light imaging of a living animal of embodiment 5, including the steps of:

the screening of the pharmacological effects of several drugs developed against a certain target strain will be described as an example.

Step 1, integral optical detection: carrying out fluorescence labeling (such as fluorescent protein labeling) on a target strain acted by a medicament to be selected, then injecting the strain into a small living animal body, fixing the small living animal body on the animal fixing component 3 after a certain time, closing the box door 23, and ensuring a dark environment in the dark box 2; the temperature control component 5 works to provide a proper temperature environment through air heating, the first rotary driving mechanism 4 works to drive the small living animal to rotate at a constant speed, the exciting light source 24 works to emit exciting light to irradiate the small living animal, the detection lens 26 of the electron multiplying CCD10 collects fluorescence emitted by the small living animal, and the average value of fluorescence data collected in unit rotation number or unit time is used as a primary detection result;

step 2, taking down the whole animal fixing component 3, injecting a research and development medicine to the living small animal fixed on the animal fixing component, carrying out fluorescence detection according to the same method as the step 1 after a certain time, and comparing detection results of two times, so as to quickly screen out the medicine with a certain drug effect; in the embodiment, when the drug has the drug effect on the target strain, the target strain can be killed or destroyed to quench or reduce the intensity of fluorescence generated by excitation, so that the drug effect can be preliminarily judged according to the difference of the fluorescence intensity of two detection results;

step 3, local optical detection: when the research and development of the drug 1 is found to have a certain drug effect through screening in the step 2, and a deep screening example is needed (for example, whether the drug effect can reach the expectation or not, differences of the effects of the drug in each main organ and the like are judged), local optical detection is needed:

step 4, after finding that the medicine 1 has a certain efficacy in the step 2, carrying out local optical detection according to the following method:

A. injecting a target strain into a living small animal body, carrying out integral optical detection after a certain time, recording a result, observing a main part of the strain infected by the small animal, and taking the part as an interested part of local optical detection;

B. the electron multiplication CCD10 and the exciting light source 24 stop working first, the first rotary driving mechanism 4 drives the animal fixing component 3 to rotate to a proper position to facilitate the local optical detection of the interested part of the living small animal, the three-dimensional displacement mechanism 6 and the second rotary driving mechanism 64 work to make the probe 250 of the PMT detector 25 to be in place and close to the interested part (in the process, the door of the box can be kept closed, the operation can be carried out by the guidance of an infrared camera, at the moment, the probe does not start working, and the door of the box can also be opened to position the probe); then under the condition of ensuring the dark environment in the camera bellows 2, the excitation light source 24 works, the three-dimensional displacement mechanism 6 and the second rotary driving mechanism 64 work, so that the probe 250 of the PMT detector 25 moves and clings to the interested part to carry out local optical detection, in the process, the position and the angle of the probe 250 can be controlled to carry out small-range adjustment so as to fully collect fluorescence emitted by the interested part, and the maximum value in the adjustment process can be taken as a measurement result so as to reduce the measurement error caused by the position factor of the probe 250;

C. the drug 1 was injected into the body of a small living animal, the detection was performed according to the method of step A, B, the result obtained at step A, B was compared with the result obtained at step C, and the drug effect of the developed drug 1 could be demonstrated by accurate data based on the comparison of fluorescence intensity or photon amount.

Of course, the device of the present invention is not limited to the above method, and may be used in combination with other methods for drug screening. For example, compared with the drug 1 with drug effect, the drug 1 can be subjected to fluorescence labeling (fluorescent dye labeling method or quantum dot labeling method, etc.), then the drug 1 is injected into the body of a small living animal, and the device of the invention is used for carrying out overall optical detection and local optical detection, so that the specific distribution and metabolic condition of the drug in the animal can be observed, and richer experimental data can be provided for drug screening.

While embodiments of the invention have been disclosed above, it is not limited to the applications listed in the description and the embodiments, which are fully applicable in all kinds of fields of application of the invention, and further modifications may readily be effected by those skilled in the art, so that the invention is not limited to the specific details without departing from the general concept defined by the claims and the scope of equivalents.

Claims

1. A drug screening device based on in vivo visible light imaging of a living animal, comprising: the drug screening device comprises an equipment box (1), a camera box (2), an electron multiplying CCD (10) arranged in the equipment box (1), an excitation light source (24), a PMT detector (25), a detection lens (26) and a camera (27) arranged in the camera box (2), wherein the detection lens (26) is connected with the electron multiplying CCD (10), and the drug screening device is characterized by further comprising an animal fixing component (3) rotatably arranged in the camera box (2), a first rotation driving mechanism (4) used for driving the animal fixing component (3) to rotate around a Y axis, a temperature control component (5) used for providing wind warm for a living animal fixed on the animal fixing component (3) and a three-dimensional displacement mechanism (6) arranged in the camera box (2);

the field of view of the detection lens (26) covers the whole living animal on the animal fixing component (3) so as to carry out whole optical detection on the living animal;

the probe (250) of the PMT detector (25) is arranged at the driving end of the three-dimensional displacement mechanism (6), and the three-dimensional displacement mechanism (6) drives the probe (250) to move in XYZ three axes so as to perform local optical detection on the living animal on the animal fixing component (3).

2. The drug screening device based on in vivo visible light imaging of the living animal according to claim 1, wherein the animal fixing component (3) comprises a support rod (30) connected to the bottom surface of the top plate (21) of the dark box (2), a rotating seat (31) rotatably connected to the inner wall of the side plate (20) of the dark box (2), and an animal fixing frame (32) with one end rotatably connected to the support rod (30) and the other end drivingly connected to the rotating seat (31), the first rotating driving mechanism (4) drives the rotating seat (31) to rotate so as to drive the animal fixing frame (32) to rotate;

the animal fixing frame (32) comprises a main beam (320) arranged along the X direction, a head fixing plate (321) slidably arranged on the main beam (320), a tail fixing plate (322) slidably arranged on the main beam (320), an auxiliary fixing beam (323) connected to the main beam (320) and arranged along the Y direction, a first joint (324) fixedly connected to a first end of the main beam (320) and a second joint (325) fixedly connected to a second end of the main beam (320);

the bottom of the supporting rod (30) is connected with a mounting seat (33), and the first joint (324) is rotatably arranged on the mounting seat (33);

the rotating seat (31) is provided with a driving jack (310) for the second connector (325) to be inserted in a matched mode, and the second connector (325) is in driving connection with the rotating seat (31) through a key groove structure.

3. The drug screening apparatus based on in vivo visible light imaging of living animal as claimed in claim 2, wherein the first rotary driving mechanism (4) comprises a first motor (40) disposed on the outer wall of the side plate (20) of the camera bellows (2), a driving gear (42) in driving connection with the output shaft of the first motor (40), and a driven gear (43) fixedly sleeved on the rotary base (31) and engaged with the driving gear (42).

4. The drug screening device based on in vivo visible light imaging of a living animal according to claim 3, wherein the mounting seat (33) comprises a main mounting seat (330) fixedly connected to the bottom of the support rod (30), a main mounting hole (331) formed in the main mounting seat (330) along the X direction, a sub-mounting seat (332) slidably disposed in the main mounting hole (331) along the X direction, a sub-mounting hole (333) formed in the sub-mounting seat (332) along the X direction, and a compression spring (334) connected between the inner wall of the main mounting hole (331) and the sub-mounting seat (332);

the inner wall of the main mounting hole (331) is also provided with a limiting guide groove (335) along the X direction, and the outer wall of the sub-mounting seat (332) is connected with a limiting guide block (336) which is slidably arranged in the limiting guide groove (335);

the first joint (324) is rotatably disposed in the sub-mounting hole (333).

5. The drug screening device based on in-vivo visible light imaging of the living animal according to claim 2, wherein the main beam (320), the auxiliary fixing beam (323) and the second joint (325) are provided with cavities (326) inside, the surfaces of the main beam (320) and the auxiliary fixing beam (323) are provided with air outlets (327) communicated with the cavities (326), and the end of the cavity (326) of the second joint (325) is provided with a bell mouth (3260);

the animal fixing frame is characterized in that an air supply opening (200) is formed in a side plate (20) of the camera bellows (2), an air supply channel (314) communicated with the air supply opening (200) and the driving insertion hole (310) is formed in the rotary seat (31), the animal fixing frame (32) is connected to the rotary seat (31), and the air supply opening (200), the air supply channel (314), the horn mouth (3260) and the cavity (326) are communicated in sequence.

6. The drug screening device based on in vivo visible light imaging of living animals according to claim 5, wherein the temperature control assembly (5) comprises a sealing cover (50) connected to a side plate (20) of the camera bellows (2), a fan (51) disposed inside the sealing cover (50), a heater (52) disposed inside the air supply outlet (200), and a temperature sensor (53) disposed inside the air supply channel (314);

the first motor (40) is arranged inside the sealing cover (50), an air inlet (500) is formed in the sealing cover (50), the fan (51) is arranged below the first motor (40), the air supply outlet (200) is positioned below the first motor (40), and an arc-shaped air deflector (501) is arranged below the inside of the sealing cover (50);

under the action of the fan (51), air outside the sealing cover (50) enters the sealing cover (50) through the air inlet (500), then is blown to the first motor (40), then enters the cavity (326) after sequentially passing through the air supply outlet (200), the air supply channel (314) and the horn mouth (3260), and finally is discharged from the main beam (320) and the air outlet (327) on the auxiliary fixing beam (323) and blown to the living animals fixed on the animal fixing frame (32).

7. The drug screening device based on in-vivo visible light imaging of the living animal according to claim 1, characterized in that a reflector (28) is further arranged on the bottom plate (22) of the dark box (2), the reflector (28) is arranged below the animal fixing frame (32), and a box door (23) is arranged at the front part of the dark box (2);

the equipment box is characterized in that a main control module (11) and a power supply module (12) are further arranged in the equipment box (1), and a touch display screen (13) is arranged on a front panel of the equipment box (1).

8. The drug screening device based on in vivo visible light imaging of the living animal according to claim 1, characterized in that a ventilation port (201) is arranged on a side plate (20) of the dark box (2), a filter (202) is arranged at the inner end of the ventilation port (201), and a light blocking assembly (7) is arranged at the outer end of the ventilation port (201);

the light blocking assembly (7) comprises a light blocking block (70) connected to a side plate (20) of the dark box (2), a ventilation channel (71) arranged in the light blocking block (70) and communicated with the ventilation port (201), and a light shield (72) arranged on the light blocking block (70);

the air exchange channel (71) comprises an inner port (710) communicated with the air exchange port (201), a U-shaped channel (711) communicated with the inner port (710), and an outer port (712) communicated with the U-shaped channel (711), wherein the inner port (710) and the outer port (712) are conical ports with large outer parts and small inner parts;

the light shield (72) is arranged above the outer port (712) in a covering mode, the light shield (72) comprises a light shield cylinder (720) connected with the light blocking block (70) and a sealing cover plate (721) connected to the light shield cylinder (720), grid holes (722) are formed in the light shield cylinder (720), and the air exchange channel (71) is communicated with the external environment through the grid holes (722).

9. The drug screening apparatus based on in vivo visible light imaging of a living animal according to claim 7, wherein the three-dimensional displacement mechanism (6) comprises a Y-direction displacement mechanism (60) disposed at a side portion of the reflector (28), an X-direction displacement mechanism (61) disposed on the Y-direction displacement mechanism (60), a Z-direction displacement mechanism (62) disposed on the X-direction displacement mechanism (61), and a driving block (63) disposed on the Z-direction displacement mechanism (62), wherein the driving block (63) can perform XYZ three-direction movements under the driving of the Y-direction displacement mechanism (60), the X-direction displacement mechanism (61), and the Z-direction displacement mechanism (62);

the driving block (63) is also connected with a second rotary driving mechanism (64), the probe (250) is arranged on the second rotary driving mechanism (64), and the rotary mechanism is used for driving the probe (250) to rotate around an X axis;

the second rotary driving mechanism (64) comprises a second motor (640) arranged on the driving block (63) and a rotary block (641) in driving connection with an output shaft of the second motor (640), and the probe (250) is connected to the rotary block (641).

10. The drug screening device based on in-vivo visible light imaging of the living animal according to claim 9, wherein the rotation block (641) comprises a main plate body (642) in driving connection with the output shaft of the second motor (640) and a buffer plate body (643) connected with the back surface of the main plate body (642), the probe (250) is connected to the buffer plate body (643);

the back of the main plate body (642) is provided with an installation groove (6420), a plurality of guide rods (6421) are arranged in the installation groove (6420), the buffer plate body (643) is connected with a sliding connecting block (6430) which is inserted in the mounting groove (6420) in a matching way, the sliding connecting block (6430) is provided with a guide hole (6431) for the guide rod (6421) to be inserted in a matching way, the upper part and the lower part of the guide rod (6421) are respectively sleeved with an upper buffer spring (6422) and a lower buffer spring (6423), the upper buffering spring (6422) is positioned between the upper inner wall of the mounting groove (6420) and the upper surface of the sliding connection block (6430), the lower buffer spring (6423) is positioned between the bottom inner wall of the mounting groove (6420) and the lower surface of the sliding connection block (6430), the sliding connection block (6430) can slide back and forth relative to the guide bar (6421).