CN111239995A - Microscopic imaging system for cell typing - Google Patents
Microscopic imaging system for cell typing Download PDFInfo
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- CN111239995A CN111239995A CN202010229805.9A CN202010229805A CN111239995A CN 111239995 A CN111239995 A CN 111239995A CN 202010229805 A CN202010229805 A CN 202010229805A CN 111239995 A CN111239995 A CN 111239995A
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- 238000003384 imaging method Methods 0.000 title claims abstract description 33
- 238000006073 displacement reaction Methods 0.000 claims description 36
- 230000005284 excitation Effects 0.000 claims description 10
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- 238000005859 coupling reaction Methods 0.000 claims description 8
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- 230000033001 locomotion Effects 0.000 abstract description 6
- 238000000799 fluorescence microscopy Methods 0.000 abstract description 4
- 210000004027 cell Anatomy 0.000 description 64
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- 238000004458 analytical method Methods 0.000 description 4
- 210000000601 blood cell Anatomy 0.000 description 3
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- 230000009286 beneficial effect Effects 0.000 description 1
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- 239000008280 blood Substances 0.000 description 1
- 238000004820 blood count Methods 0.000 description 1
- 239000006227 byproduct Substances 0.000 description 1
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- 238000003745 diagnosis Methods 0.000 description 1
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- 230000000694 effects Effects 0.000 description 1
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- 238000001917 fluorescence detection Methods 0.000 description 1
- 239000007850 fluorescent dye Substances 0.000 description 1
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- G—PHYSICS
- G02—OPTICS
- G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
- G02B21/00—Microscopes
- G02B21/0004—Microscopes specially adapted for specific applications
- G02B21/002—Scanning microscopes
- G02B21/0024—Confocal scanning microscopes (CSOMs) or confocal "macroscopes"; Accessories which are not restricted to use with CSOMs, e.g. sample holders
- G02B21/0052—Optical details of the image generation
- G02B21/0076—Optical details of the image generation arrangements using fluorescence or luminescence
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- G—PHYSICS
- G02—OPTICS
- G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
- G02B21/00—Microscopes
- G02B21/0004—Microscopes specially adapted for specific applications
- G02B21/002—Scanning microscopes
- G02B21/0024—Confocal scanning microscopes (CSOMs) or confocal "macroscopes"; Accessories which are not restricted to use with CSOMs, e.g. sample holders
- G02B21/0036—Scanning details, e.g. scanning stages
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- G—PHYSICS
- G02—OPTICS
- G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
- G02B21/00—Microscopes
- G02B21/06—Means for illuminating specimens
- G02B21/08—Condensers
- G02B21/12—Condensers affording bright-field illumination
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- G—PHYSICS
- G02—OPTICS
- G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
- G02B21/00—Microscopes
- G02B21/24—Base structure
- G02B21/241—Devices for focusing
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- G—PHYSICS
- G02—OPTICS
- G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
- G02B21/00—Microscopes
- G02B21/24—Base structure
- G02B21/26—Stages; Adjusting means therefor
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- G—PHYSICS
- G02—OPTICS
- G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
- G02B21/00—Microscopes
- G02B21/34—Microscope slides, e.g. mounting specimens on microscope slides
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- Analytical Chemistry (AREA)
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- Optics & Photonics (AREA)
- Microscoopes, Condenser (AREA)
- Investigating, Analyzing Materials By Fluorescence Or Luminescence (AREA)
Abstract
The invention discloses a microscopic imaging system for cell typing, which comprises: the system comprises a white light source, a first collimating lens, a three-dimensional moving objective table, an objective lens group, a fluorescent module group, a second collimating lens, a camera and a computer; the three-dimensional moving object stage is used for bearing a sample and realizing the position adjustment of the sample in the XYZ three directions; the fluorescent module group comprises a plurality of groups of fluorescent modules, and each fluorescent module comprises a fluorescent light source, a third collimating lens, an exciting light filter, a dichroic mirror and a fluorescent light filter. The microscopic imaging system for cell typing can perform bright field imaging and fluorescence imaging, and obtain the image of a cell sample to be tested for realizing cell typing; according to the invention, the three-dimensional moving object stage is arranged to realize the position adjustment of the sample in the three directions of XYZ, the cell sample is scanned through the position adjustment in the XY directions, the images of all the cell samples in the sample pool can be obtained, and the focusing of the sample can be realized through the movement in the Z direction.
Description
Technical Field
The invention relates to the field of microscopic imaging, in particular to a microscopic imaging system for cell typing.
Background
Clinical blood cell typing analysis comprises cell subset classification counting and cell morphology detection, and has great significance in early disease discovery, especially in diagnosis of blood system diseases.
A blood cell counting instrument is a commonly used full-automatic blood cell analyzer, at present, China is basically monopolized by products of foreign enterprises such as BECKMAN, Yapek, Bayer, Xisenmeikang, Japan photoelectricity and the like, and in recent years, a BC-5500 white blood cell classification instrument is also provided by domestic Mirui company. The instrument mainly counts and analyzes various types of cells and does not have the function of cell morphology detection.
The cytomorphology examination is mainly completed by cell staining and microscope examination at present, clinical blood cell analysis does not meet simple cell counting, but rather tends to analyze morphological characteristics such as internal DNA, RNA, immunity and the like of various cells and subgroups thereof, and can meet the detection requirements of clinical automation, batch, standardization and traceability.
Aiming at the requirement of accurate cell typing analysis of clinical cells, the development of an imaging system which can be used for cell typing is of great significance. For example, patent 201910390572.8 discloses a compact fluorescence inverted microscopy imaging system, which can count and classify cells and achieve the purpose of cell detection and classification by integrating a bright field imaging mode and a fluorescence imaging mode. The method is characterized in that a slide glass needs to do two-dimensional motion in the XY direction to obtain an image of a complete slide glass, but a specific scheme for realizing the XY direction movement of the slide glass is not disclosed, in addition, the focusing is realized by moving an objective lens along the Z direction, so that the objective lens linear switching focusing mechanism not only needs to realize the switching of an objective lens in and out an optical path, but also needs to realize the focusing in the Z direction, and the complexity of the device is easily increased.
Disclosure of Invention
The technical problem to be solved by the present invention is to provide a microscopic imaging system for cell typing, which is aimed at the above-mentioned deficiencies in the prior art.
In order to solve the technical problems, the invention adopts the technical scheme that: a microscopic imaging system for cell typing, comprising: the system comprises a white light source, a first collimating lens, a three-dimensional moving objective table, an objective lens group, a fluorescent module group, a second collimating lens, a camera and a computer;
the three-dimensional moving object stage is used for bearing a sample and realizing the position adjustment of the sample in the three directions of XYZ;
the fluorescence module group comprises a plurality of groups of fluorescence modules, and each fluorescence module comprises a fluorescence light source, a third collimating lens, an excitation light filter, a dichroic mirror and a fluorescence filter;
the light emitted by the white light source is collimated by the first collimating lens and then irradiates a sample on the three-dimensional moving objective table, is focused by an objective lens in the objective lens group, and then is reflected by the dichroic mirror, passes through the fluorescent filter and the second collimating lens in sequence and reaches the camera;
the light emitted by the fluorescent light source sequentially passes through the third collimating lens, the excitation light filter, the dichroic mirror for transmission and the objective lens in the objective lens group and then irradiates on the sample, and the fluorescence generated by the sample after being excited sequentially passes through the objective lens in the objective lens group, the dichroic mirror for reflection, the fluorescent light filter for filtration and the second collimating lens and then reaches the camera.
Preferably, the objective lens group comprises three objective lenses with magnification of 5, 10 and 20 times respectively, which can be switched into the optical path.
Preferably, the set of fluorescence modules comprises three sets capable of emitting excitation light of different wavelengths that can be switched into the optical path.
Preferably, a sample cell for holding a sample is arranged on the three-dimensional moving object stage.
Preferably, the three-dimensional moving stage includes a base plate, an X displacement mechanism provided on the base plate, a Z displacement mechanism provided on the X displacement mechanism, a Y displacement mechanism provided on the Z displacement mechanism, and a stage provided on the Y displacement mechanism, and the sample cell is provided on the stage.
Preferably, the X displacement mechanism comprises two sets of X slide rails arranged in parallel along the X direction on the bottom plate, an X slide plate arranged on the two sets of X slide rails in a sliding manner by matching with the X slide block, an X threaded sliding sleeve fixedly connected to the bottom of the X slide plate, an X motor fixedly connected to the bottom plate, and an X lead screw arranged in the X threaded sliding sleeve in a driving and matching manner and connected to the X motor.
Preferably, the Z displacement mechanism includes a guide bracket fixedly connected to the X sliding plate, a guide slide bar fixedly connected to the guide bracket, a Z motor fixedly connected to the X sliding plate, a Z lead screw drivingly connected to the Z motor, and a Z sliding plate slidably disposed on the guide slide bar;
but Z slide last rigid coupling have the slidable cover establish Z sliding sleeve on the direction slide bar and with Z lead screw thread fit's Z screw thread sliding sleeve.
Preferably, the Y displacement mechanism comprises a support fixedly connected to the Z sliding plate, a Y sliding rail arranged on the support along the Y direction, a Y sliding block slidably arranged on the Y sliding rail, a Y motor fixedly connected to the support, and a Y lead screw which is in driving connection with the Y motor and is arranged in a Y threaded hole formed in the Y sliding block in a matched and penetrating manner;
the bottom of the Y sliding block is provided with a Y sliding groove matched with the Y sliding rail, and the carrying seat is fixedly connected to the Y sliding block.
Preferably, a cover plate groove and a mounting groove for accommodating the sample cell are sequentially formed in the object carrying seat from the surface to the bottom at one end far away from the X motor, a cover plate rotatably connected with the object carrying seat is arranged in the cover plate groove in a matched manner, a first light transmitting opening is formed in the middle of the cover plate, and gaskets are arranged at two ends of the bottom surface of the cover plate in the Y direction;
a second light-transmitting opening is formed in the bottom surface of the mounting groove;
the lateral part of carrying the thing seat and being in the apron groove still is provided with and is used for fixing the retaining member of apron, the retaining member includes the rigid coupling and is in hollow pillar on carrying the thing seat, slidable insert and establish bolt in the pinhole seted up in the hollow pillar, connect extension spring, rigid coupling between the bottom surface of bolt bottom and pinhole are in the drawing disk and the rigid coupling on bolt top are in the drawing disk lateral part be used for the gland the preforming of apron.
Preferably, the bottom surface of the mounting groove is connected with a support plate through a spring, and the support plate is provided with a third light-transmitting opening;
first light trap, second light trap, third light trap link up in proper order, first light trap allows the light irradiation of top incidence to insert and establish in the sample cell in the mounting groove, second light trap, third light trap allow the light that the sample cell sent to jet out in proper order downwards.
The invention has the beneficial effects that: the microscopic imaging system for cell typing can perform bright field imaging and fluorescence imaging, and obtain the image of a cell sample to be tested for realizing cell typing; according to the invention, the position adjustment of the sample in the three directions of XYZ can be realized by arranging the three-dimensional moving object stage, the cell sample is scanned by the position adjustment in the XY direction, the images of all the cell samples in the sample pool can be obtained, and the focusing of the sample can be realized by the movement in the Z direction, so that the objective lens group does not need to have the function of displacement in the Z direction, the displacement requirement in the XYZ direction is integrated into the three-dimensional moving object, and the arrangement of each mechanism of the system is facilitated and the simplification of the whole structure is realized.
Drawings
FIG. 1 is a schematic diagram of a microscopic imaging system for cell typing according to the present invention;
FIG. 2 is a schematic structural diagram of a three-dimensional movable stage according to the present invention;
FIG. 3 is a schematic view of the structure of the carrier of the present invention (not loaded into a sample well);
FIG. 4 is a schematic diagram of the structure of a sample cell according to the present invention;
FIG. 5 is a schematic view of the structure of the carrier of the present invention (loaded into a sample cell);
FIG. 6 is a schematic view of the bottom surface of the cover plate according to the present invention;
fig. 7 is a schematic view of the carrier of the present invention (with the cover plate removed);
fig. 8 is a schematic view showing the structure of the locker of the present invention.
Description of reference numerals:
1-a white light source; 2-a first collimating lens; 3-three-dimensional moving object stage; 4-an objective lens group; 5-a fluorescence module group; 6-a second collimating lens; 7-a camera; 8, a computer; 9-a sample cell;
30-a bottom plate;
31-X displacement mechanism; 310-X slide; 311-X slider; 312-X slide; 313-X thread sliding bush; 314-X motor; 315 — X lead screw;
32-Z displacement mechanism; 320, a guide bracket; 321-guiding the sliding rod; 322-Z motor; 323-Z lead screw; 324-Z sled; 325-Z sliding sleeve; 326-Z threaded sliding sleeves;
33-Y displacement mechanism; 330-a support; 331-Y slide rail; 332 — Y slider; 333-Y motor; 334-Y lead screw; 335-Y threaded hole; 336-Y chute; 337-Y bearing;
34-carrying seat; 340-cover plate groove; 341-mounting groove; 342-a cover plate; 343-a first light transmission opening; 344 — a gasket; 345-second light-transmitting opening; 346-a retaining member; 347-support plate; 348 — a spring; 349 — third light transmission opening; 3460 hollow pillar; 3461-bolt; 3462 tension spring; 3463-pulling the disc; 3464 tabletting; 3465 pinhole; 3466-guide sleeve;
50-a fluorescent light source; 51-a third collimating lens; 52-excitation light filter; 53-dichroic mirror; 54-a fluorescent filter; 55-fluorescent module switching mechanism;
90-a sample chamber; 91-pool surface.
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.
As shown in fig. 1, a microscopic imaging system for cell typing of the present embodiment includes: the three-dimensional moving objective table comprises a white light source 1, a first collimating lens 2, a three-dimensional moving objective table 3, an objective lens group 4, a fluorescent module group 5, a second collimating lens 6, a camera 7 and a computer 8;
the three-dimensional moving object stage 3 is used for bearing a sample and realizing the position adjustment of the sample in three directions of XYZ;
the fluorescent module group 5 includes a plurality of groups of fluorescent modules, and each fluorescent module includes a fluorescent light source 50, a third collimating lens 51, an excitation light filter 52, a dichroic mirror 53, and a fluorescent filter 54.
The invention can carry out bright field imaging (counting) and fluorescence imaging (morphological detection), and specifically comprises the following steps:
when bright field imaging is carried out, light emitted by a white light source 1 is collimated by a first collimating lens 2 and then uniformly irradiated onto a cell sample in a sample cell 9 on a three-dimensional moving object stage 3, then the light is focused by an objective lens in an objective lens group 4, and then the light is reflected by a dichroic mirror 53, focused by a fluorescent light filter 54 and a second collimating lens 6 in sequence and then reaches a camera 7;
the light emitted by the fluorescence light source 50 sequentially passes through the third collimating lens 51, the excitation light filter 52, the dichroic mirror 53 for transmission, and the objective lens in the objective lens group 4 to irradiate the sample, and the fluorescence generated by the excitation of the dye in the cell sample sequentially passes through the objective lens in the objective lens group 4, the dichroic mirror 53 for reflection, the fluorescence filter 54 for filtration, and the second collimating lens 6 for focusing, and then reaches the camera 7.
Wherein, the cell sample to be detected is dyed by specific fluorescent dye in advance according to the detection requirement, and then is added into the sample cell 9 to form a single-layer cell tiling state.
In order to capture all cell sample pictures, the three-dimensional moving object stage 3 can drive the sample cell 9 to perform XY two-dimensional movement so as to automatically scan the cell sample pictures; meanwhile, the sample pool 9 is driven by the three-dimensional moving object stage 3 to move in the Z direction, so that the automatic focusing of the sample can be realized, and a clear picture can be obtained.
In the present embodiment, the camera 7 is preferably a low-noise CMOS camera.
In this embodiment, the objective lens group 4 includes three objective lenses capable of switching to enter the optical path, which are 5 × objective lens (5 times), 10 × objective lens (10 times) and 20 × objective lens (20 times), respectively, and is used for meeting the imaging requirements of different cell samples, and certainly, in order to meet more requirements, other objective lenses with more times can be adopted. The three objective lenses can be switched by a conventional objective lens switching mechanism (without having a focusing function), such as an objective lens switcher.
In this embodiment, the fluorescence module group 5 includes three groups capable of switching to enter the light path and emitting fluorescence with different wavelengths, and different fluorescence light sources 50 emit excitation light with different wavelengths, such as UV LED, blue LED, green LED, and the like, thereby realizing multicolor fluorescence detection. The fluorescence module switching mechanism 55 can be manually or electrically switched by a fluorescence module switching mechanism 55, and the fluorescence module switching mechanism 55 can be a conventional product, for example, patent 201910390572.8, a fluorescence module switching mechanism used in a compact fluorescence inverted microscopic imaging system.
In this embodiment, the computer 8 and the camera 7 acquire an image captured by the camera 7. In the subsequent processing, the computer 8 processes the image, and finally realizes the rapid typing of the cell to be detected by analyzing the morphology and the fluorescence characteristic of the cell. It should be understood that the present invention mainly improves the structure of the whole imaging system, and the subsequent analysis and processing of the image only needs to use the conventional technology, which is not the focus of the present application and the solution to be protected, so that the detailed description is not needed in the present application.
The three-dimensional moving object stage 3 is provided with a sample cell 9 for containing samples, and the three-dimensional moving object stage 3 drives the sample cell 9 to perform three-dimensional movement so as to realize operations such as scanning and focusing. Referring to fig. 2-7, in a preferred embodiment, the three-dimensional moving stage 3 employs the following scheme.
The three-dimensional moving stage 3 includes a base plate 30, an X displacement mechanism 31 provided on the base plate 30, a Z displacement mechanism 32 provided on the X displacement mechanism 31, a Y displacement mechanism 33 provided on the Z displacement mechanism 32, and a stage 34 provided on the Y displacement mechanism 33, and the sample cell 9 is provided on the stage 34.
The X displacement mechanism 31 includes two sets of X slide rails 310 arranged in parallel on the bottom plate 30 along the X direction, an X slide plate 312 slidably arranged on the two sets of X slide rails 310 through the cooperation of an X slide block 311, an X threaded slide sleeve 313 fixedly connected to the bottom of the X slide plate 312, an X motor 314 fixedly connected to the bottom plate 30, and an X lead screw 315 drivingly connected to the X motor 314 and fittingly inserted into the X threaded slide sleeve 313.
Two sets of X slide rails 310 are respectively provided with 2X slide blocks 311, the X thread sliding sleeve 313 is internally provided with a thread hole, and an X screw 315 is in thread fit with the X thread sliding sleeve 313. The X sliding plate 312 is further provided with an X bearing (not shown in fig. 2) engaged with the left end of the X lead screw 315. The X motor 314 drives the X screw rod 315 to rotate, thereby driving the X threaded sliding sleeve 313 to slide along the X direction, so that the X sliding plate 312 can move along the X direction as a whole.
The Z displacement mechanism 32 includes a guide bracket 320 fixedly connected to the X sliding plate 312, a guide sliding rod 321 fixedly connected to the guide bracket 320, a Z motor 322 fixedly connected to the X sliding plate 312, a Z lead screw 323 drivingly connected to the Z motor 322, and a Z sliding plate 324 slidably disposed on the guide sliding rod 321; a Z sliding sleeve 325 which can be sleeved on the guide sliding rod 321 in a sliding manner and a Z threaded sliding sleeve 326 which is in threaded fit with the Z screw rod 323 are fixedly connected to the Z sliding plate 324. The support 330 is further provided with a Y bearing 337 engaged with the end of the Y screw 334.
An unthreaded hole is formed in the Z sliding sleeve 325, and the guide sliding rod 321 can be slidably inserted into the unthreaded hole to guide the Z sliding plate 324, so that the linearity of the Z sliding plate moving along the Z direction is ensured. In this embodiment, the guide bracket 320, the guide sliding rod 321, and the Z sliding sleeve 325 are two sets, and are symmetrically disposed on two sides of the Z lead screw 323, so as to ensure a good guiding effect.
A threaded hole is formed in the Z threaded sliding sleeve 326, the Z lead screw 323 is in threaded fit with the Z threaded sliding sleeve 326, and the Z motor 322 drives the Z lead screw 323 to rotate, so that the Z threaded sliding sleeve 326 is driven to slide along the Z direction, and the Z sliding plate 324 can integrally move along the Z direction.
The Y displacement mechanism 33 includes a support 330 fixedly connected to the Z sliding plate 324, a Y sliding rail 331 arranged on the support 330 along the Y direction, a Y sliding block 332 slidably arranged on the Y sliding rail 331, a Y motor 333 fixedly connected to the support 330, and a Y lead screw 334 drivingly connected to the Y motor 333 and fittingly arranged in a Y threaded hole 335 formed in the Y sliding block 332;
the bottom of the Y slider 332 is provided with a Y chute 336 for matching with the Y slide rail 331, and the object holder 34 is fixedly connected to the Y slider 332.
The Y screw 334 is in threaded fit with the Y threaded hole 335, and the Y motor 333 drives the Y screw 334 to rotate, thereby driving the Y slider 332 to slide along the Y direction, so that the object carrying seat 34 can move along the Y direction.
The position of the sample on the object stage 34 in the three-dimensional direction can be adjusted by the X displacement mechanism 31, the Z displacement mechanism 32 and the Y displacement mechanism 33, so as to realize operations such as automatic scanning and automatic focusing, and realize an imaging function. In a preferred embodiment, the X motor 314, the Z motor 322 and the Y motor 333 are high-precision stepping motors, so that the precision of displacement adjustment is ensured.
The object carrying seat 34 is used for carrying the sample cell 9, and since the object carrying seat 34 needs to move in three-dimensional directions, the object carrying seat 34 needs to be capable of fixing the sample cell 9 well without affecting the imaging of the sample. In a preferred embodiment, the carrier 34 is configured as follows.
A cover plate 342 groove 340 and a mounting groove 341 for accommodating the sample cell 9 are sequentially formed in the object holder 34 from the surface to the bottom at the end far away from the X motor 314, the cover plate 342 rotatably connected with the object holder 34 is arranged in the cover plate 342 groove 340 in a matching manner, a first light-transmitting opening 343 is formed in the middle of the cover plate 342, and gaskets 344 are arranged at two ends of the bottom surface of the cover plate 342 in the Y direction; the left end of the cover plate 342 is rotatably connected to the carrier 34 by a hinge or a pin. The pad 344 is made of soft material such as rubber or sponge to protect the surface of the sample cell 9.
The bottom surface of the mounting groove 341 is provided with a second light-transmitting opening 345;
a locking member 346 for fixing the cover plate 342 is further disposed on the object holder 34 at a side portion of the groove 340 of the cover plate 342, the locking member 346 includes a hollow pillar 3460 fixedly attached to the object holder 34, a bolt 3461 slidably inserted into a pin hole 3465 formed in the hollow pillar 3460, a tension spring 3462 connected between a bottom end of the bolt 3461 and a bottom surface of the pin hole 3465, a pull plate 3463 fixedly attached to a top end of the bolt 3461, and a pressing piece 3464 fixedly attached to a side portion of the pull plate 3463 for pressing the cover plate 342. In a more preferred embodiment, a guide sleeve 3466 is further disposed between the latch 3461 and the pin hole 3465, and the latch 3461 can slide freely up and down relative to the guide sleeve 3466.
In a further preferred embodiment, a supporting plate 347 is connected to the bottom surface of the mounting groove 341 through a spring 348, the spring 348 includes a plurality of springs arranged at intervals to ensure that the supporting plate 347 can provide a stable support for the sample cell 9, and the supporting plate 347 is provided with a third light-transmitting opening 349 (not shown);
the first light-transmitting opening 343, the second light-transmitting opening 345 and the third light-transmitting opening 349 are sequentially penetrated, the first light-transmitting opening 343 allows light incident from above to irradiate the sample cell 9 inserted in the mounting groove 341, and the second light-transmitting opening 345 and the third light-transmitting opening 349 allow light emitted from the sample cell 9 to sequentially emit downward.
Referring to fig. 2, the left end of the object carrying seat 34 extends out of the support 330, and the mounting groove 341 is located at the left end of the object carrying seat 34, so as to ensure that the view below the mounting groove 341 is clear, so as to conveniently arrange the objective lens group 4 and the fluorescent module, and enable light passing through the mounting groove 341 to propagate downwards and smoothly pass through the objective lens group 4 and the fluorescent module.
When the sample cell 9 is installed in the installation groove 341, the pull disc 3463 is pulled to separate the pressing piece 3464 from the cover plate 342, then the pull disc 3463 is rotated (the bolt 3461 is connected with the bottom surface of the pin hole 3465 through the tension spring 3462, the bolt 3461 has a certain rotational freedom degree relative to the hollow support 3460 and can rotate at least 90 °), the upper part of the cover plate 342 is deviated, then the cover plate 342 is opened, and the sample cell 9 is inserted into the installation groove 341; then the cover plate 342 is covered, the pull plate 3463 is rotated to reset, the pull plate 3463 is released, the latch 3461 is inserted into the hollow column 3460 again by the pull of the pull spring 3462, and the pressing piece 3464 presses the cover plate 342. The gasket 344 on the bottom surface of the cover plate 342 is pressed against the two ends of the sample cell 9 along the Y direction, and the spring 348 under the support plate 347 generates an upward pressing force, so as to generate a vertical clamping force on the sample cell 9, so that the sample cell 9 can be firmly fixed in the mounting groove 341, and the sample cell 9 is prevented from sliding when the position of the loading base 34 is adjusted. It should be understood that, in the present embodiment, the sample cell 9 has a rectangular shape, and referring to fig. 4, the gasket 344 is pressed against the cell surface 91 at both ends of the sample cell 9, so as not to cover the sample cavity 90 in the sample cell 9, and not to block the light entering into the sample cell 9.
In the description of the present invention, it is to be understood that the terms "central," "longitudinal," "lateral," "length," "width," "thickness," "upper," "lower," "front," "rear," "left," "right," "vertical," "horizontal," "top," "bottom," "inner," "outer," "clockwise," "counterclockwise," "axial," "radial," "circumferential," and the like are used in the orientations and positional relationships indicated in the drawings for convenience in describing the invention and to simplify the description, and are not intended to indicate or imply that the referenced devices or elements must have a particular orientation, be constructed and operated in a particular orientation, and are therefore not to be considered limiting of the invention.
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 (10)
1. A microscopic imaging system for cell typing, comprising: the system comprises a white light source, a first collimating lens, a three-dimensional moving objective table, an objective lens group, a fluorescent module group, a second collimating lens, a camera and a computer;
the three-dimensional moving object stage is used for bearing a sample and realizing the position adjustment of the sample in the three directions of XYZ;
the fluorescence module group comprises a plurality of groups of fluorescence modules, and each fluorescence module comprises a fluorescence light source, a third collimating lens, an excitation light filter, a dichroic mirror and a fluorescence filter;
the light emitted by the white light source is collimated by the first collimating lens and then irradiates a sample on the three-dimensional moving objective table, is focused by an objective lens in the objective lens group, and then is reflected by the dichroic mirror, passes through the fluorescent filter and the second collimating lens in sequence and reaches the camera;
the light emitted by the fluorescent light source sequentially passes through the third collimating lens, the excitation light filter, the dichroic mirror for transmission and the objective lens in the objective lens group and then irradiates on the sample, and the fluorescence generated by the sample after being excited sequentially passes through the objective lens in the objective lens group, the dichroic mirror for reflection, the fluorescent light filter for filtration and the second collimating lens and then reaches the camera.
2. A microscopic imaging system for cell typing according to claim 1, wherein said objective lens group comprises three objective lenses with 5, 10 and 20 magnifications switchable into the optical path.
3. A microscopic imaging system for cell typing according to claim 2, wherein said fluorescence module set comprises three sets capable of emitting excitation light of different wavelengths switchable into optical paths.
4. A microscopic imaging system for cell typing according to claim 1, wherein a sample cell for holding a sample is disposed on said three-dimensional moving stage.
5. The microscopic imaging system for cell typing according to claim 4, wherein the three-dimensional moving stage comprises a base plate, an X displacement mechanism provided on the base plate, a Z displacement mechanism provided on the X displacement mechanism, a Y displacement mechanism provided on the Z displacement mechanism, and a stage provided on the Y displacement mechanism, the sample cell being provided on the stage.
6. The microscopic imaging system for cell typing according to claim 5, wherein said X displacement mechanism comprises two sets of X slide rails disposed in parallel along the X direction on said bottom plate, an X slide plate slidably disposed on said two sets of X slide rails by means of X slide block fit, an X thread sliding sleeve fixedly connected to the bottom of said X slide plate, an X motor fixedly connected to said bottom plate, and an X lead screw drivingly connected to said X motor and fittingly inserted in said X thread sliding sleeve.
7. The microscopic imaging system for cell typing according to claim 6, wherein the Z displacement mechanism comprises a guide bracket fixedly connected to the X slide plate, a guide slide rod fixedly connected to the guide bracket, a Z motor fixedly connected to the X slide plate, a Z lead screw in driving connection with the Z motor, and a Z slide plate slidably arranged on the guide slide rod;
but Z slide last rigid coupling have the slidable cover establish Z sliding sleeve on the direction slide bar and with Z lead screw thread fit's Z screw thread sliding sleeve.
8. The microscopic imaging system for cell typing according to claim 7, wherein the Y displacement mechanism comprises a support fixedly connected to the Z sliding plate, a Y sliding rail arranged on the support along the Y direction, a Y sliding block slidably arranged on the Y sliding rail, a Y motor fixedly connected to the support, and a Y lead screw which is in driving connection with the Y motor and is fittingly arranged in a Y threaded hole formed in the Y sliding block;
the bottom of the Y sliding block is provided with a Y sliding groove matched with the Y sliding rail, and the carrying seat is fixedly connected to the Y sliding block.
9. The microscopic imaging system for cell typing according to claim 8, wherein a cover plate groove and a mounting groove for accommodating the sample cell are sequentially formed in the object holder from the surface to the bottom at the end away from the X motor, a cover plate rotatably connected with the object holder is arranged in the cover plate groove in a matching manner, a first light-transmitting opening is formed in the middle of the cover plate, and gaskets are arranged at two ends of the bottom surface of the cover plate in the Y direction;
a second light-transmitting opening is formed in the bottom surface of the mounting groove;
the lateral part of carrying the thing seat and being in the apron groove still is provided with and is used for fixing the retaining member of apron, the retaining member includes the rigid coupling and is in hollow pillar on carrying the thing seat, slidable insert and establish bolt in the pinhole seted up in the hollow pillar, connect extension spring, rigid coupling between the bottom surface of bolt bottom and pinhole are in the drawing disk and the rigid coupling on bolt top are in the drawing disk lateral part be used for the gland the preforming of apron.
10. The microscopic imaging system for cell typing according to claim 9, wherein a support plate is connected to the bottom surface of the mounting groove through a spring, and a third light-transmitting opening is formed in the support plate;
first light trap, second light trap, third light trap link up in proper order, first light trap allows the light irradiation of top incidence to insert and establish in the sample cell in the mounting groove, second light trap, third light trap allow the light that the sample cell sent to jet out in proper order downwards.
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| CN202010229805.9A CN111239995A (en) | 2020-03-27 | 2020-03-27 | Microscopic imaging system for cell typing |
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