CN110353720B - Synchrotron radiation X-ray micro CT and fixing device for living body imaging of small animal limbs - Google Patents
Synchrotron radiation X-ray micro CT and fixing device for living body imaging of small animal limbs Download PDFInfo
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- CN110353720B CN110353720B CN201910807991.7A CN201910807991A CN110353720B CN 110353720 B CN110353720 B CN 110353720B CN 201910807991 A CN201910807991 A CN 201910807991A CN 110353720 B CN110353720 B CN 110353720B
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- 241001465754 Metazoa Species 0.000 title claims abstract description 68
- 230000005469 synchrotron radiation Effects 0.000 title claims abstract description 34
- 238000003384 imaging method Methods 0.000 title claims description 29
- 206010002091 Anaesthesia Diseases 0.000 claims abstract description 4
- 230000037005 anaesthesia Effects 0.000 claims abstract description 4
- 210000003414 extremity Anatomy 0.000 claims description 35
- 238000010603 microCT Methods 0.000 claims description 13
- 230000005855 radiation Effects 0.000 claims description 5
- 241000283973 Oryctolagus cuniculus Species 0.000 claims description 3
- 210000003205 muscle Anatomy 0.000 claims description 3
- 238000012800 visualization Methods 0.000 claims description 3
- 210000000689 upper leg Anatomy 0.000 claims description 2
- 238000011503 in vivo imaging Methods 0.000 claims 5
- 238000010859 live-cell imaging Methods 0.000 claims 1
- 230000001360 synchronised effect Effects 0.000 claims 1
- 230000004083 survival effect Effects 0.000 abstract description 5
- 230000015572 biosynthetic process Effects 0.000 abstract description 3
- 238000002474 experimental method Methods 0.000 abstract description 2
- 230000007774 longterm Effects 0.000 abstract description 2
- 210000002414 leg Anatomy 0.000 description 8
- 210000000544 articulatio talocruralis Anatomy 0.000 description 7
- 210000000988 bone and bone Anatomy 0.000 description 5
- 238000000034 method Methods 0.000 description 5
- 210000001503 joint Anatomy 0.000 description 4
- 241000699666 Mus <mouse, genus> Species 0.000 description 3
- 230000008569 process Effects 0.000 description 3
- 230000001105 regulatory effect Effects 0.000 description 3
- 210000004872 soft tissue Anatomy 0.000 description 3
- 238000013170 computed tomography imaging Methods 0.000 description 2
- 238000010586 diagram Methods 0.000 description 2
- 230000035876 healing Effects 0.000 description 2
- 210000001519 tissue Anatomy 0.000 description 2
- 208000025978 Athletic injury Diseases 0.000 description 1
- 206010061225 Limb injury Diseases 0.000 description 1
- 241000699670 Mus sp. Species 0.000 description 1
- 241000283984 Rodentia Species 0.000 description 1
- 208000026137 Soft tissue injury Diseases 0.000 description 1
- 206010041738 Sports injury Diseases 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 210000003423 ankle Anatomy 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000008827 biological function Effects 0.000 description 1
- 244000309466 calf Species 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 239000013013 elastic material Substances 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- 230000004907 flux Effects 0.000 description 1
- 210000002683 foot Anatomy 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 210000003141 lower extremity Anatomy 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 230000000149 penetrating effect Effects 0.000 description 1
- 230000001681 protective effect Effects 0.000 description 1
- 230000008929 regeneration Effects 0.000 description 1
- 238000011069 regeneration method Methods 0.000 description 1
- 230000008439 repair process Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
- A61B6/00—Apparatus or devices for radiation diagnosis; Apparatus or devices for radiation diagnosis combined with radiation therapy equipment
- A61B6/02—Arrangements for diagnosis sequentially in different planes; Stereoscopic radiation diagnosis
- A61B6/03—Computed tomography [CT]
- A61B6/032—Transmission computed tomography [CT]
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
- A61B6/00—Apparatus or devices for radiation diagnosis; Apparatus or devices for radiation diagnosis combined with radiation therapy equipment
- A61B6/04—Positioning of patients; Tiltable beds or the like
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
- A61B6/00—Apparatus or devices for radiation diagnosis; Apparatus or devices for radiation diagnosis combined with radiation therapy equipment
- A61B6/10—Safety means specially adapted therefor
- A61B6/107—Protection against radiation, e.g. shielding
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61B—DIAGNOSIS; SURGERY; IDENTIFICATION
- A61B2503/00—Evaluating a particular growth phase or type of persons or animals
- A61B2503/40—Animals
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- High Energy & Nuclear Physics (AREA)
- Veterinary Medicine (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Public Health (AREA)
- Pathology (AREA)
- Radiology & Medical Imaging (AREA)
- Physics & Mathematics (AREA)
- Heart & Thoracic Surgery (AREA)
- Molecular Biology (AREA)
- Surgery (AREA)
- Animal Behavior & Ethology (AREA)
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Abstract
The utility model provides a synchrotron radiation X-ray micro CT and fixing device suitable for little animal four limbs live body formation of image, fixing device is including being used for placing the objective table of little animal after anesthesia, fix stand and the lid dress on the objective table, with objective table top and the confined plumbous lid all around, synchrotron radiation light source includes fixing device, fixing device's objective table fixes on the rotatory sample stage of synchrotron radiation X-ray micro CT, and sleeve axle center and rotatory sample stage rotation axle center coaxial setting on the objective table, this fixing device of device can wholly and locally fix living animal, can avoid the trunk position outside the four limbs joint to receive the influence of high-dose ray simultaneously, and the survival of the living animal of maximum protection is convenient for carry out long-term animal experiment.
Description
Technical Field
The invention relates to a synchrotron radiation X-ray micro CT and fixing device suitable for living body imaging of limbs of small animals.
Background
The sports injury has the characteristics of high recurrence rate, difficult regeneration and repair and the like, and mainly occurs in bones and joints of limbs, and is often accompanied with soft tissue injury. Imaging evaluation is an important means for researching healing of limb injury of small animals, and currently common imaging means comprise micro CT, MRI and the like, but are limited by spatial resolution, and three-dimensional imaging under ultra-high resolution is difficult to realize by conventional means.
With the advent of synchrotron radiation X-ray micro-CT (Synchrotron radiation micro, CT, SR, μCT), the high luminous flux, high collimation and high coherence properties of the composition can be used for three-dimensional imaging of tissue microscopic morphology structures, and the resolution can reach submicron level. The three-dimensional microstructure of the bones, joints and soft tissues of the limbs is analyzed by utilizing the synchrotron radiation technology, so that the method has important research significance. There is no method or technical parameter for imaging the limbs of small animals in vivo in the disclosed invention.
Moreover, due to the large SR and μCT radiation doses, the survival of small animals is affected, and the animals often need to be sacrificed and isolated tissues are imaged, so that the real-time healing change process in the animals cannot be dynamically observed. Conventional small animal fixing devices generally fix and brake a target area by means of suspension or clamping, have no specific device for fixing limbs and have no protective device for high radiation dose, so the conventional small animal fixing devices are not suitable for imaging of synchrotron radiation small animals, and the small animal fixing devices for imaging of synchrotron radiation living animals are not disclosed in the disclosed invention.
Disclosure of Invention
Aiming at the defects of the prior art, the invention provides a synchrotron radiation X-ray micro CT and a fixing device which are specially used for the living body imaging of the limbs of the small animals, can correspondingly adjust the fixing position according to the size of the animal body, can prevent the trunk part from being irradiated by high-dose rays, and ensure the survival of the small animals after the synchrotron radiation scanning, and are suitable for the living body imaging of the limbs of the small animals.
The aim of the invention is achieved by adopting the following technical scheme:
the utility model provides a fixing device suitable for live body formation of image of toy limbs, includes the objective table that is used for placing the toy after anesthesia, fixes stand and the lid on the objective table, with objective table top and the sealed plumbous lid all around, four the stand distributes around the toy and corresponds with the position of toy limbs, wherein be fixed with the rope that is used for fixing toy limbs on three stands, be fixed with the sleeve of perpendicular setting on another stand through the fixation clamp, the inside hollow formation of sleeve supplies toy to wait to image limbs to pass, and with its cavity that fixes a position, plumbous lid is equipped with the through-hole that matches with sleeve size in sleeve corresponding position, plumbous lid is adorned on the objective table back, the sleeve stretches out plumbous lid, and the location of above-mentioned sleeve to wait to image limbs means: the diameter of the cavity is larger than the diameter of the bones of the limb to be imaged of the small animal, but slightly smaller than the outer diameter of the muscles of the limb to be imaged, so that the limb to be imaged is clamped when passing through.
Further, be equipped with a plurality of through-holes on the objective table and constitute the stand fixed orifices that is used for fixed stand, stand fixed orifices inner wall is the helicitic texture, the bottom outer wall of stand is equipped with stand fixed orifices internal thread assorted external screw thread.
Further, the fixation clamp includes stand clamping part and sleeve clamping part, stand clamping part is connected with sleeve clamping part, and stand clamping part includes a width and stand diameter assorted recess, stand fixation nut is installed to recess one side, the stand is fixed the stand in the recess through stand fixation nut after the stand cartridge is in the recess, sleeve clamping part is a clamp, the elasticity of clamp through sleeve fixation nut regulation centre gripping, the sleeve is installed in the clamp, thereby fixes the sleeve in the clamp through screwing sleeve fixation nut.
Furthermore, the cavity of the sleeve forms a round table with small top and large bottom, and the inner wall of the opening at the top of the cavity is provided with an elastic ring for clamping the muscles of the calf or thigh of the small animal.
The synchrotron radiation X-ray micro CT suitable for the living body imaging of the limbs of the small animals comprises the fixing device, an objective table of the fixing device is fixed on a synchrotron radiation light source rotary sample table, and the axis of a sleeve on the objective table is coaxially arranged with the rotary axis of the rotary sample table.
Further, specific parameters of the synchrotron radiation light source are set, the energy of the light source is set to 15keV, the effective imaging resolution of the CCD detector is set to 3.25um, and the exposure time is set to 100ms.
Further, the high resolution camera is set to capture 1080 original tomo images, 5 dark images and 18 flat and gap images.
Furthermore, PITRE software adopted by the synchrotron radiation light source is set for reconstructing the slices of the data, and VG studio3.0 software is adopted for three-dimensional visualization of the reconstructed data.
Further, the distance between the rotation axis and the CCD detector was set to 10cm.
Further, the small animal is a mouse, a rat, a rabbit, or the like.
By adopting the structure, the invention has the following advantages:
1) The invention provides a special fixing device for limb living body imaging of a small animal aiming at synchrotron radiation X-ray micro CT imaging, which can be used for integrally and locally fixing the living small animal, correspondingly adjusting the position of a fixing column according to the size of the animal body, avoiding the influence of large-dose radiation irradiation on the trunk part, protecting the survival of the small animal to the greatest extent and facilitating long-term animal experiments;
2) The invention realizes high-precision imaging of bones, joints and soft tissues of limbs by using specific imaging parameters, has higher imaging resolution compared with micro CT imaging, has clearer microstructure display, is beneficial to further analyzing three-dimensional microstructure characteristics of bones, joints and soft tissues of limbs, and deeply understand the relationship between the structure and biological functions of a motion system.
Drawings
FIG. 1 is a schematic view of a fixing device according to the present invention;
FIG. 2 is a schematic view of the lead cover of the present invention;
FIG. 3 is a schematic structural view of the column of the present invention;
FIG. 4 is a schematic view of the structure of the fixing clip of the present invention;
FIG. 5 is a schematic structural diagram of synchrotron radiation X-ray micro CT;
fig. 6 is a graph comparing the results of synchrotron radiation X-ray micro-CT and micro-CT scanning.
In the figure: 1. an objective table; 2. a column; 3. a sleeve; 4. a fixing clamp; 5. a lead cover; 6. a column fixing hole; 7. a fixing portion; 8. a threaded portion; 9. a through hole; 10. a column fixing nut; 11. a column clamping portion; 12. a sleeve fixing nut; 13. a sleeve clamping portion; 14. a light source; 15. a monochromator; 16. rotating the sample stage; 17. a CCD detector.
Detailed Description
Embodiments of a synchrotron radiation light source and a fixing device suitable for living body imaging of a small animal limb are described in detail below with reference to the accompanying drawings and specific examples.
As shown in fig. 1 to 6, a synchrotron radiation X-ray micro CT suitable for imaging a limb living body of a small animal comprises an objective table 1 fixed on a synchrotron radiation X-ray micro CT rotary sample table 16, the objective table 1 is used for placing a small animal after anesthesia, in this embodiment, the small animal refers to a rodent, preferably a mouse, a rat or a rabbit, the objective table 1 is of a rectangular plate-shaped structure, a plurality of upright fixing holes 6 are arrayed on the objective table 1, the upright fixing holes 6 are through holes, the inner wall of the upright fixing holes 6 are of a threaded structure, four uprights 2 are connected in the upright fixing holes 6 of the objective table 1 in a threaded manner, after the small animal is placed on the objective table 1, the four uprights 2 are installed around the small animal, three uprights are connected with three supporting feet of the small animal through ropes, so that the limb of the small animal is prevented from moving in the imaging process, a fixing clamp 4 is fixed on another upright, the fixing clamp 4 is used for fixing a sleeve 3, the sleeve 3 is stably placed in a vertical shape, a fourth supporting foot for synchrotron radiation scanning penetrates from the sleeve 3 from the inside from the upper side, the four supporting feet are threaded through the sleeve 3, the four supporting feet are threaded through the four supporting feet 5, and a proper lead cover is extended out of the four supporting feet 5, and a proper lead cover is fixed on the four supporting feet 5 is extended out of the four supporting feet 5, and a proper lead cover is fixed.
Fig. 2, 3 and 4 are detailed views of the object stage, the upright, the sleeve, the fixing clamp and the lead cover, wherein the upright fixing hole 6 is used for fixing the upright 2 on one hand and realizing the circulation of internal and external air through the upright fixing hole 6 after the lead cover 5 is covered on the other hand, so as to ensure the survival of animals to be imaged. The upright post 2 is of a cylindrical structure and consists of a fixing part 7 and a bottom thread part 8. The fixing part 7 is used for fixing ropes to be connected with limbs of the small animals, the fixing clamp 4 can be installed at the same time, the thread part 8 is matched with the thread structure on the inner surface of the stand column fixing hole 6 of the objective table, and the stand column 2 can adjust corresponding fixing positions according to the sizes of the small animals. The fixing clamp 4 comprises a column clamping part 11 and a sleeve clamping part 13, the column clamping part 11 is connected with the sleeve clamping part 13, the column clamping part 11 is a groove, a column fixing nut 10 is arranged on one side of the groove, after a column is inserted into the groove, the column fixing nut 10 fixes the column in the groove, the sleeve clamping part 13 is a clamp, the clamp is tightly regulated by the sleeve fixing nut 12, the sleeve 3 is arranged in the clamp, the sleeve fixing nut 12 is screwed to fix the sleeve 3 in the clamp, and the height of the fixing clamp on the column 2 is regulated to fix the scanned fourth support leg on one hand and prevent the scanned fourth support leg from shaking, and on the other hand, the position of the sleeve 3 is regulated to guide the small animal foot to pass through the sleeve 3, so that the ankle joint is exposed from the top of the sleeve 3, and the subsequent fourth support leg is conveniently exposed from the through hole 9 on the lead cover 5.
The sleeve 3 is of a hollow round platform structure, the sleeve is divided into different models of height and inner diameter according to the size of a small animal, an elastic ring which is used for the small animal leg to pass through and is convenient to clamp the small animal leg is arranged on the inner wall of the opening at the top of the sleeve, the elastic ring is made of an elastic material, the opening of the elastic ring is slightly smaller than the small animal leg diameter, when the small animal leg is clamped into the sleeve 3, the small animal leg is partially fixed by the elastic ring at the top of the sleeve 3, the small animal ankle joint extends out of the sleeve 3, the small animal ankle joint is prevented from moving in the imaging process by the elastic ring, the small animal ankle joint is ensured to be fixed, and the sleeve 3 is fixed to the small animal ankle joint through the sleeve clamping part 13 of the fixing clamp 4.
The lead cover 5 is a cover-shaped structure made of lead material, and can prevent rays from penetrating. The lead cover 5 is provided with a through hole 9 on the surface, and when the lead cover 5 is used for covering the trunk part of the small animal, the lower limb to be scanned can extend out of the through hole 9 for scanning.
In synchrotron radiation imaging, as shown in fig. 5, the synchrotron radiation light source includes a light source 14, a monochromator 15, a rotary sample stage 16 and a CCD detector 17, the light source 14 is used for providing parallel light with energy of 8.0 and 72.5kev, the monochromator 15 is used for converting the light into a high coherence monochromatic light beam, the rotary sample stage 16 is fixed at the downstream of the light source 14, and the CCD detector 17 is located on a regulatable guide rail. After the mice were anesthetized, they were properly fixed on the stage 1, with the fixation ensuring that the scan site was at the center of rotation of the rotating sample stage 16, to ensure the highest imaging quality and relatively lowest radiation dose, the following scan parameters were used: the energy of the light source 14 was set to 15kev, the effective imaging resolution of the ccd detector 17 was set to 3.25um, the sample-to-detector distance was set to 10cm (with both phase contrast and absorption contrast), the exposure time was set to 100ms (reduced scan time), 1080 original tomo images were captured using a high resolution camera, 5 dark images, and 18 flat, gap images. The adopted PITRE software is used for reconstructing the slices of the data, and VG studio3.0 software is used for carrying out three-dimensional visualization on the reconstructed data.
FIG. 6 is a graph comparing microCT and SR [ mu ] CT images of the ankle joint of a mouse, wherein A in FIG. 6 is a cross-sectional and loss-of-shape contrast graph; FIG. 6B is a gray value distribution diagram of a scribing region; FIG. 6C is a schematic view showing the structure of the SR [ mu ] CT ankle joint loss-form surface. It can be seen that SR μct shows more clearly ankle details than microCT imaging.
The foregoing is a preferred embodiment of the invention, but it will be understood by those skilled in the art that various changes in form and details may be made therein without departing from the spirit and scope of the invention as defined by the appended claims.
Claims (8)
1. A fixing device suitable for live imaging of toy limbs, characterized in that: the device comprises an object table (1) for placing a small animal after anesthesia, upright posts fixed on the object table (1) and a lead cover (5) for covering the upper part and the periphery of the object table (1), wherein four upright posts are distributed on the periphery of the small animal and correspond to the positions of the limbs of the small animal, three upright posts are fixedly provided with ropes for fixing the limbs of the small animal, the other upright post is fixedly provided with a vertically arranged sleeve through a fixing clamp (4), a cavity for the small animal to pass through and locate the limbs to be imaged is formed in the sleeve, the inner wall of an opening at the top of the cavity is provided with an elastic ring for clamping the muscles of the small animal's lower leg or thigh, the lead cover (5) is provided with a through hole matched with the sleeve in size at the position corresponding to the sleeve, and the lead cover (5) is covered on the object table (1) and then extends out of the lead cover (5);
the object stage (1) is provided with a plurality of through holes to form an upright post fixing hole (6) for fixing an upright post, the inner wall of the upright post fixing hole (6) is of a threaded structure, and the outer wall of the bottom of the upright post is provided with external threads matched with the internal threads of the upright post fixing hole (6);
the fixing clamp is characterized in that the fixing clamp (4) comprises a column clamping part (11) and a sleeve clamping part (13), the column clamping part (11) is connected with the sleeve clamping part (13), the column clamping part (11) comprises a groove with the width matched with the diameter of the column, a column fixing nut (10) is arranged on one side of the groove, the column is fixed in the groove through the column fixing nut (10) after being inserted into the groove, the sleeve clamping part (13) is a clamp, the clamp adjusts clamping tightness through the sleeve fixing nut (12), the sleeve is arranged in the clamp, and the sleeve is fixed in the clamp through screwing the sleeve fixing nut (12).
2. The fixation device for in vivo imaging of small animal extremities as claimed in claim 1, wherein: the cavity of the sleeve forms a round table with small top and large bottom.
3. Synchrotron radiation X-ray micro-CT suitable for the living imaging of small animal limbs, comprising a fixing device according to claim 1 or 2, characterized in that: the object stage (1) of the fixing device is fixed on a synchronous radiation X-ray micro CT rotary sample stage (16), and the axis of a sleeve on the object stage (1) is coaxial with the rotary axis of the rotary sample stage (16).
4. Synchrotron radiation X-ray micro-CT suitable for the living imaging of small animal limbs according to claim 3, characterized in that: specific parameters of the synchrotron radiation X-ray micro CT are set, the energy of the light source (14) is set to 15keV, the effective imaging resolution of the CCD detector (17) is set to 3.25um, and the exposure time is set to 100ms.
5. Synchrotron radiation X-ray micro-CT suitable for in vivo imaging of small animal limbs according to claim 4, characterized in that: setting a high-resolution camera to capture 1080 original tomo images, 5 dark images and 18 flat and gap images.
6. Synchrotron radiation X-ray micro-CT suitable for in vivo imaging of small animal limbs according to claim 4, characterized in that: and setting PITRE software adopted by synchrotron radiation X-ray micro CT to reconstruct the data in a slicing way, and adopting VG studio3.0 software to perform three-dimensional visualization on the reconstructed data.
7. Synchrotron radiation X-ray micro-CT suitable for in vivo imaging of small animal limbs according to claim 4, characterized in that: the distance between the rotation axis and the CCD detector (17) is set to be 10cm.
8. Synchrotron radiation X-ray micro-CT suitable for in vivo imaging of small animal limbs according to claim 5, characterized in that: the small animal is a mouse, a rat or a rabbit.
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| CN110974283A (en) * | 2019-12-20 | 2020-04-10 | 中南大学湘雅二医院 | Fixator for measuring rat by bone densitometer by dual-energy X-ray absorption method and manufacturing method |
| CN111419181B (en) * | 2020-03-31 | 2023-07-25 | 浙江大学 | Device for long-time living body imaging of small animals and application method thereof |
| CN111820873A (en) * | 2020-03-31 | 2020-10-27 | 同济大学 | Living mouse muscle confocal imaging chamber |
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