CN209004307U - A kind of fixation device of the living small animal window for being imaged under inverted fluorescence microscope - Google Patents
A kind of fixation device of the living small animal window for being imaged under inverted fluorescence microscope Download PDFInfo
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- CN209004307U CN209004307U CN201721874911.2U CN201721874911U CN209004307U CN 209004307 U CN209004307 U CN 209004307U CN 201721874911 U CN201721874911 U CN 201721874911U CN 209004307 U CN209004307 U CN 209004307U
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Abstract
The fixation device for the living small animal window that the utility model relates to a kind of for being imaged under inverted fluorescence microscope, window is offered at living small animal tissue or organ to be seen, the fixation device includes pedestal (1) and the bogey (3) for loading living small animal, and bogey is removably attachable on pedestal；The opening (3-1) for fixing living small animal tissue or organ window is offered on bogey, and imaging window (1-1) corresponding with bogey is offered on pedestal and is imaged for living small animal.Living animal imaging window is fixed with can be convenient by the fixed device, fixed device is highly suitable for inverted fluorescence microscope and carries out imaging to organ, tissue of living small animal etc., toy, which is fixed, using the fixation device can make toy have better comfort level, can carry out observational study to the different parts of toy for more time, more efficiently.
Description
Technical field
The utility model relates to Instrumentation Engineering technologies, and in particular, to a kind of for being imaged under inverted fluorescence microscope
The fixation device of living small animal window.
Background technique
In past 20 years, the development of fluorescin technology and intravital microscope technology expands us to single thin
The understanding of born of the same parents' information, so that us also be promoted to step a huge step to the understanding of cell biology.These are mostly known
Knowing is mainly to be obtained by cell injuring model, and the purposes in terms of in vitro culture experimental implementation is very large.However, they are simultaneously
The true representations that cannot be learned entirely as complicated multicellular organism.Nematode imaging method develops into solution this problem
The first step, it provides the genetic manipulation easily performed and therefore discloses several physiology courses for occurring in tissue.Recently, light
The technology for learning microscope imaging has spread in living mammal model, thus also develops a kind of referred to as intravital microscope
New imaging method.Rapidly, living imaging has become the important tool in several fields, such as Neurobiology, immunology
And oncobiology.
In addition, the success of living cells imaging, which opens, studied cell under molecular level and physiological condition in past 5 years
The gate of biology.By improve microscope, for imaging analysis optics and software and can control over time and space
The novel genetic tool for making the expression of given gene makes it possible that these improve.Particularly, several expression fluorescent marker eggs
Knocking in mouse and obtaining availability with relatively low cost for white matter, makes scientist be able to solve the science of several keys
Problem.However, this technology cannot achieve repetition imaging, or even when animal organism is strictly measured and is controlled, living body at
As that can only carry out a few houres.In order to which imaging time is extended to several days or some months, even more long, living imaging window technique is obtained
Development is arrived.By the imaging window, the imaging of cellular level can be carried out to living animal, and can be to each in living tissue
Cell type, the cellular structures factor (such as: extracellular matrix), fluorescent marker protein or biological sensor can carry out can
Depending on the dynamic studies changed.By utilizing imaging window, living tissue can be repeated to be imaged, the visualization time of tissue extends to
Several days or some months, more even up to 1 year.Animal can restore from anesthesia during imaging, avoid because anesthesia duration is too long
And the death rate is caused to increase.And the use of imaging window realizes the imaging repeatedly to deep tissues in animal body, passes through hand
Art exposure imaging position, and then visualize the abdomen organs such as small intestine, kidney, spleen, pancreas and liver.
It is since mouse position is easy to happen greatly to imaging window observation with inverted fluorescence microscope in viviperception
The variation of amplitude, furthermore, very big variation may will occur for the data of the living imaging in a flash of variation, it is some have it is great
The discovery of meaning is often flooded wherein.Therefore, excellent in living imaging window with inverted fluorescence microscope to play
Gesture, stable imaging window maintain to be a technical problem to be solved urgently with fixation.
Utility model content
The living small animal window that the purpose of the utility model is to provide a kind of for being imaged under inverted fluorescence microscope
Fixed device, the fixation device are convenient to that living small animal is fixed, and it is aobvious to be conveniently used in inversion fluorescence
The imaging of living small animal window in micro mirror.
The utility model provides a kind of fixation device of living small animal window for being imaged under inverted fluorescence microscope,
Window is offered at living small animal tissue or organ to be seen, which includes pedestal and for loading living body petty action
The bogey of object, the bogey are removably attachable on the pedestal；It is offered on the bogey for solid
Determine the opening of living small animal tissue or organ window, imaging window corresponding with the bogey is offered on the pedestal
It is imaged for living small animal.
Optionally, the opening on the bogey, the living small animal tissue or organ window and described
The imaging window of pedestal is circle.
Optionally, the bogey is hollow cylindrical drum.
Optionally, the fixed device further includes fixed bracket, and the fixed bracket is connected on the pedestal, described to hold
It carries to set and be removably attachable on the fixed bracket.
Optionally, the fixed bracket is two and is connected to the two sides of the imaging window on the pedestal, institute
The connecting hole for offering on fixed bracket and can accommodating the bogey end is stated, the bogey is removably attachable to institute
It states between two fixed brackets.
Optionally, perforation is offered on the fixed bracket, the fixed bracket further includes screw, and screw is removably worn
The bogey is fixed in the support bracket fastened connecting hole after crossing the perforation.
Optionally, the diameter phase of the opening on the bogey and the living small animal tissue or organ window
Together；The diameter of the imaging window of the pedestal is greater than the diameter of the living small animal tissue or organ window.
Optionally, the diameter of the imaging window of the pedestal and the living small animal tissue or the diameter of organ window
Ratio is (1.1-3): 1.
Living animal imaging window is fixed with can be convenient by the fixation device of the utility model, fixed device
It is highly suitable for inverted fluorescence microscope and living body window imaging is carried out to organ, tissue of living small animal etc., use
The comfort level of toy can be increased after the fixed device is fixed to the living body window of toy, and can effectively be controlled
Position during small animal imaging can be more convenient, the longer time, more efficiently observe the different parts of toy
Research.
Other feature and advantage of the utility model will the following detailed description will be given in the detailed implementation section.
Detailed description of the invention
Attached drawing is to be used to provide a further understanding of the present invention, and constitute part of specification, and following
Specific embodiment be used to explain the utility model together, but do not constitute limitations of the present invention.In the accompanying drawings:
Fig. 1 is the fixation device of the living small animal window for being imaged under inverted fluorescence microscope of the utility model
A kind of structural schematic diagram of specific embodiment.
Description of symbols
1 pedestal 2 fixes 3 bogey of bracket
4 screw 1-1 imaging window 2-1 connecting holes
2-2 perforation 3-1 opening
Specific embodiment
Specific embodiment of the present utility model is described in detail below in conjunction with attached drawing.It should be understood that herein
Described specific embodiment is only used for describing and explaining the present invention, and is not intended to limit the utility model.
In the present invention, in the absence of explanation to the contrary, the noun of locality used such as " upper and lower, " typically refers to
The fixation device of living small animal window for being imaged under inverted fluorescence microscope is undefined in normal operating condition, specifically
It can be found in attached page shown in FIG. 1, " inside and outside " refers to the inside and outside of respective profile.
The utility model provides a kind of fixation device of living small animal window for being imaged under inverted fluorescence microscope,
Window is offered at living small animal tissue or organ to be seen, as shown in Figure 1, the fixation device can have including 1 He of pedestal
For loading the bogey 3 of living small animal, the bogey 3 can be removably attachable on the pedestal 1；It is described
The opening 3-1 for fixing living small animal tissue or organ window can be offered on bogey 3, it can on the pedestal 1
It is imaged with the corresponding imaging window 1-1 of the bogey 3 for living small animal with offering.
The window that living small animal opens up can be the edge of a knife that opens up on living small animal to expose toy
Organ or tissue etc. facilitates observational study in turn, and the kidney of small animal living body can be specifically observed by living small animal window
Dirty, spleen, small intestine, pancreas and brain etc..
As shown in Figure 1, the opening 3-1, the living small animal tissue or organ window on the bogey 3 with
And the imaging window 1-1 of the pedestal 1 can be circle.
As shown in Figure 1, the bogey 3 can be hollow cylindrical drum.
Preferably, the internal diameter of the cylindrical drum can be slightly larger than the size of living small animal to be seen, hollow cylindrical tube edge
The both ends of length direction can open up outlet, and wherein the outlet of one end can be used for fixing and exposing the head of living small animal
Portion, the outlet of the other end can be used for fixing and exposing the tail portion of living small animal, and such toy is in carrying to the carrying
It still is able to carry out breathing well when device and tail swing, toy has better comfort level in bogey, more
Be conducive to the health of toy and the smooth development of imaging.
As shown in Figure 1, the fixed device can also include fixed bracket 2, the fixed bracket 2 be can connect in described
On pedestal 1, the bogey 3 can be removably attachable on the fixed bracket 2.
The fixed bracket 2 can be fixedly connected on pedestal 1, or can be detachably connected on pedestal 1；It is preferred that
Ground, position of the fixed bracket 2 on the pedestal 1 are adjustable.
As shown in Figure 1, the fixed bracket 2 can be two and can be connected to the imaging on the pedestal 1
The two sides of window 1-1 can offer the connecting hole 2-1 that can accommodate 3 end of bogey, institute on the fixed bracket 2
Stating bogey 3 can be removably attachable between described two fixed brackets 2.
The fixed bracket 2 can be detachably connected on pedestal 1, and link position of the fixed bracket 2 on pedestal 1 can
With adjustable, in this way, be adjusted to can be convenient just can be with the bogey 3 for the distance between two fixed brackets 2
Length match, the bogey 3 can be conveniently connected between two fixed brackets 2.
As shown in Figure 1, can offer perforation 2-2 on the fixed bracket 2, the fixed bracket 2 can also include spiral shell
The bogey 3 can be removably fixed on the institute of the fixed bracket 2 by nail 4, screw 4 after the perforation 2-2
It states in connecting hole 2-1.
The connection elasticity between bogey 3 and fixed bracket 2 is adjusted with can be convenient by the screw 4,
It can make to rotate bogey 3 after lightening restrictions between bogey 3 and fixed bracket 2 by rotating screw 4, can be reversed after adjusting
Rotation screw 4 makes closely to connect between bogey 3 and fixed bracket 2.
It is above-mentioned using the utility model for inverted fluorescence microscope under the fixation of living small animal window that is imaged
When device, first can by after anesthesia and opened up observation organ or tissue's window living small animal be put into the bogey
In 3, so that the opening 3-1 of living small animal window and bogey 3 is located at same position, that is, pass through the opening 3-1 of bogey 3
The organ or tissue in living small animal window be able to be observed well, meanwhile, make head and the portion of living small animal
Not Jing Guo the opening at 3 both ends of bogey be pierced by；Then, the both ends that will be mounted with the bogey 3 of living small animal are solid respectively
It is fixed into two fixed brackets 2 on pedestal 1, make living small animal window, bogey 3 be open 3-1 and pedestal imaging
Window 1-1 is just in same vertical straight line, i.e., after pedestal 1 being lifted certain altitude upwards, from the bottom up from 1 bottom of pedestal
When, living small animal window is seen by the opening 3-1 that the imaging window 1-1 of pedestal 1 can pass through bogey 3, and then can be with
The internal organ or tissue of observational study living small animal window exposure, in this process, after can suitably unclamping the screw 4
Bogey 3 is rotated, again tightens the screw 4 after adjusting to suitable position；Finally, living body petty action can will be fixed with
The fixation device of object is placed on inverted microscope objective table, can carry out imaging.Pass through the fixed dress of the utility model
The liver and kidney for setting the intravital mouse after fixing can be imaged well by inverted fluorescence microscope.
As shown in Figure 1, the opening 3-1 and the living small animal tissue or organ window on the bogey 3
Diameter can be identical；The diameter of the imaging window 1-1 of the pedestal 1 can be greater than the living small animal tissue or organ window
The diameter of mouth.
The ratio of the diameter of the imaging window 1-1 of the pedestal 1 and living small animal tissue or organ window diameter can be with
For (1.1-3): 1.
As shown in Figure 1, the ratio of the diameter of the diameter of the imaging window 1-1 of the pedestal 1 and the living small animal window
Value can be (1.1-3): 1.It is above-mentioned numerical value by the ratio of setting imaging window 1-1 diameter and living small animal window diameter
Range can observe organ, the tissue etc. that are exposed in living small animal window with more convenient operator.
Further the utility model is illustrated by the following examples, following embodiment is only used for describing and explaining this
Utility model is not intended to limit the utility model.
In the examples below, IIW refers to intravital imaging window, and used inverted microscope can be with
For Nikon inverted fluorescence microscope, model Ti-U.
Embodiment 1: application of the fixed device in mouse liver living imaging
The preparation step of mouse liver living imaging window:
1) autoclave sterilization surgical instrument uses medicinal alcohol sterile surgical platform region；
2) through chloraldurate (4%, 350mg/kg) anesthetized mice is injected intraperitoneally, after mouse anesthesia, abdomen depilation, conventional hand
Art disinfection；
3) skin is clamped using tweezers, and is cut since xiphoid-process using micro-dissections scissors, be longitudinally formed a 15mm
Midline incision, separation skin and abdominal muscles and open abdominal cavity；
4) xiphoid-process is caught with haemostatic clamp, clamped as much as possible；
5) lift haemostatic clamp in encephalic direction, meanwhile, liver is lightly wiped by using humidity (PBS) cotton swab, can be seen
To falciform ligament；
6) after using haemostatic clamp to clamp xiphoid-process after five minutes, and xiphoid-process is removed；
7) liver is stirred using cotton swab, the sky that then will be generated between the laterally inserted liver of cotton gauze and diaphragm using tweezers
In gap；
8), apart from about 1 millimeter of edge, suture is first passed through since the tail end of notch using needle thread passing stomach wall and skin
Line passes through skin, then passes through opposite stomach wall from external-to-internal.Next, moving 3mm along notch, and from inside to outside again
Stomach wall and skin are passed through by suture.It is sutured along entire edge of wound repetitive operation, eventually forms a circular seams zygonema.
In order to which the skin of suture and stomach wall to be fixed in annular groove, it not make notch too big.The last one stitch should be located at distance
At first about 3 millimeters of entrance, it is ensured that be left outside ring；
9) stomach wall and skin are carefully placed in the groove of IIW using tweezers；
10) bracing wire suture one by one, by suture tightening in the groove of IIW.
11) in the inner peripheral surface gluing of IIW, then preprepared sheet glass is tightly attached in inside groove.Fixed device
And the combination of living imaging window:
It after mouse induced anesthesia, is placed in hollow cylindrical tube, and makes the imaging window of mouse liver position body surface
It coincide with the opening of cylinder barrel, then cylindrical drum is put on V-type fixed frame, rotation cylindrical drum makes opening and the window on pedestal
It coincide, is screwed tightly, finally entire fixed device is put on inverted microscope objective table, imaging can be carried out.
Embodiment 2: application of the fixed device in mouse kidney living imaging
The preparation step of mouse kidney living imaging window:
1) autoclave sterilization surgical instrument uses medicinal alcohol sterile surgical platform region；
2) through chloraldurate (4%, 350mg/kg) anesthetized mice is injected intraperitoneally, after mouse anesthesia, back depilation, conventional hand
Art disinfection；
3) through chloraldurate (4%, 350mg/kg) anesthetized mice is injected intraperitoneally, in the middle part of back, at left of spine 0.2cm
Longitudinal incision skin (0.8-1cm) separates subcutaneous connective tissue, and detachment muscle of back enters abdominal cavity, detects and expose left kidney, can
See that cerise is presented in normal kidney；
7) kidney is stirred using cotton swab, the sky that then will be generated between the laterally inserted kidney of cotton gauze and diaphragm using tweezers
In gap；
8), apart from about 1 millimeter of edge, suture is first passed through since the tail end of notch using needle thread passing stomach wall and skin
Line passes through skin, then passes through opposite stomach wall from external-to-internal.Next, moving 3mm along notch, and from inside to outside again
Stomach wall and skin are passed through by suture.It is sutured along entire edge of wound repetitive operation, eventually forms a circular seams zygonema.
In order to which the skin of suture and stomach wall to be fixed in annular groove, it not make notch too big.The last one stitch should be located at distance
At first about 3 millimeters of entrance, it is ensured that be left outside ring；
9) stomach wall and skin are carefully placed in the groove of IIW using tweezers；
10) bracing wire suture one by one, by suture tightening in the groove of IIW；
11) in the inner peripheral surface gluing of IIW, then preprepared sheet glass is tightly attached in inside groove；Fixed device
And the combination of living imaging window:
It after mouse induced anesthesia, is placed in hollow cylindrical tube, and makes the imaging window of mouse kidney position body surface
It coincide with the opening of cylinder barrel, then cylindrical drum is put on V-type fixed frame, rotation cylindrical drum makes opening and the window on pedestal
It coincide, is screwed tightly, finally entire fixed device is put on inverted microscope objective table, imaging can be carried out.
It can be seen from above-described embodiment that the fixation device of the utility model is tieed up imaging window with can be convenient
It holds and fixes, the convenient imaging that cellular level is carried out to living animal, and can be to all types of cells, the acellular in living tissue
Structure factor (such as: extracellular matrix), fluorescent marker protein or biological sensor can carry out visual dynamic studies.
By utilizing the fixation device, living tissue can be repeated to be imaged, the visualization time of tissue extends to several days or some months,
More it is even up to 1 year.Animal can restore from anesthesia during imaging, and avoiding leads to the death rate because anesthesia duration is too long
Increase.And maintenance and fixation of the fixation device to imaging window, realize the imaging repeatedly to deep tissues in animal body,
Position is imaged by surgical exposure, and then visualizes the abdomen organs such as small intestine, kidney, spleen, pancreas and liver.
Preferred embodiments of the present invention, still, the utility model and unlimited are described in detail in conjunction with attached drawing above
Detail in above embodiment can be to the skill of the utility model in the range of the technology design of the utility model
Art scheme carries out a variety of simple variants, these simple variants belong to the protection scope of the utility model.
It is further to note that specific technical features described in the above specific embodiments, in not lance
It in the case where shield, can be combined in any appropriate way, in order to avoid unnecessary repetition, the utility model is to each
No further explanation will be given for the possible combination of kind.
In addition, any combination can also be carried out between a variety of different embodiments of the utility model, as long as it is not disobeyed
The thought for carrying on the back the utility model, equally should be considered as content disclosed in the utility model.
Claims (8)
1. a kind of fixation device of the living small animal window for being imaged under inverted fluorescence microscope, living small animal are to be seen
Tissue or organ at offer window, which is characterized in that the fixation device includes pedestal (1) and for loading living small animal
Bogey (3), the bogey (3) is removably attachable on the pedestal (1)；It is opened on the bogey (3)
Equipped with the opening (3-1) for fixing living small animal tissue or organ window, offered and the carrying on the pedestal (1)
The corresponding imaging window (1-1) of device (3) is imaged for living small animal.
2. fixed device according to claim 1, which is characterized in that the opening (3- on the bogey (3)
1), the imaging window (1-1) of the living small animal tissue or organ window and the pedestal (1) is circle.
3. fixed device according to claim 1, which is characterized in that the bogey (3) is hollow cylindrical drum.
4. fixed device according to claim 1, which is characterized in that the fixed device further includes fixed bracket (2), institute
It states fixed bracket (2) to be connected on the pedestal (1), the bogey (3) is removably attachable to the fixed bracket (2)
On.
5. fixed device according to claim 4, which is characterized in that the fixed bracket (2) is two and is separately connected
The carrying can be accommodated in offering on the two sides of imaging window (1-1) described on the pedestal (1), the fixed bracket (2)
The connecting hole (2-1) of device (3) end, the bogey (3) be removably attachable to described two fixed brackets (2) it
Between.
6. fixed device according to claim 5, which is characterized in that offer perforation (2- on the fixed bracket (2)
2), the fixed bracket (2) further includes screw (4), and screw (4) is removably by the carrying after the perforation (2-2)
Device (3) is fixed in the connecting hole (2-1) of the fixed bracket (2).
7. fixed device according to claim 2, which is characterized in that the opening (3-1) on the bogey (3)
It is identical as the diameter of the living small animal tissue or organ window；The diameter of the imaging window (1-1) of the pedestal (1) is greater than
The diameter of the living small animal tissue or organ window.
8. fixed device according to claim 7, which is characterized in that the diameter of the imaging window (1-1) of the pedestal (1)
It is (1.1-3) with the ratio of the living small animal tissue or the diameter of organ window: 1.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201721874911.2U CN209004307U (en) | 2017-12-27 | 2017-12-27 | A kind of fixation device of the living small animal window for being imaged under inverted fluorescence microscope |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201721874911.2U CN209004307U (en) | 2017-12-27 | 2017-12-27 | A kind of fixation device of the living small animal window for being imaged under inverted fluorescence microscope |
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| CN209004307U true CN209004307U (en) | 2019-06-21 |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111012524A (en) * | 2019-12-27 | 2020-04-17 | 苏州溢博伦光电仪器有限公司 | Separated optical vibration reduction platform |
| CN111419181A (en) * | 2020-03-31 | 2020-07-17 | 浙江大学 | Device for long-time living body imaging of small animal and using method thereof |
| CN111820873A (en) * | 2020-03-31 | 2020-10-27 | 同济大学 | Living mouse muscle confocal imaging chamber |
-
2017
- 2017-12-27 CN CN201721874911.2U patent/CN209004307U/en active Active
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111012524A (en) * | 2019-12-27 | 2020-04-17 | 苏州溢博伦光电仪器有限公司 | Separated optical vibration reduction platform |
| CN111419181A (en) * | 2020-03-31 | 2020-07-17 | 浙江大学 | Device for long-time living body imaging of small animal and using method thereof |
| CN111820873A (en) * | 2020-03-31 | 2020-10-27 | 同济大学 | Living mouse muscle confocal imaging chamber |
| 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 |
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