CN112327471B - Microscope system, adjusting method thereof and sequencing system - Google Patents
Microscope system, adjusting method thereof and sequencing system Download PDFInfo
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- CN112327471B CN112327471B CN201811278035.6A CN201811278035A CN112327471B CN 112327471 B CN112327471 B CN 112327471B CN 201811278035 A CN201811278035 A CN 201811278035A CN 112327471 B CN112327471 B CN 112327471B
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- G—PHYSICS
- G02—OPTICS
- G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
- G02B21/00—Microscopes
- G02B21/06—Means for illuminating specimens
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- G—PHYSICS
- G02—OPTICS
- G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
- G02B21/00—Microscopes
- G02B21/02—Objectives
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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
- G02B21/36—Microscopes arranged for photographic purposes or projection purposes or digital imaging or video purposes including associated control and data processing arrangements
- G02B21/362—Mechanical details, e.g. mountings for the camera or image sensor, housings
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Abstract
The invention discloses an adjusting method of a microscope system, which comprises the following steps: transmitting light passing through a reticle to an objective lens mounting surface by using a light source, wherein the reticle is provided with at least one identification pattern; acquiring a first image of an identification pattern formed by light reflected by an objective lens mounting surface; transmitting light passing through the reticle to the object carrying surface by using a light source; acquiring a second image of the identification pattern formed by the light reflected by the object carrying surface; and adjusting the object carrying platform to enable the second image of the identification pattern to be superposed with the first image of the identification pattern, so that the object carrying surface is parallel to the objective lens mounting surface. The adjusting method improves the efficiency of detecting whether the object carrying surface is parallel to the objective lens mounting surface. In addition, the object carrying platform is adjusted when the object carrying surface is not parallel to the objective lens mounting surface, so that the object carrying surface is parallel to the objective lens mounting surface, and the adjusting process is simple. The invention also discloses a microscope system and a sequencing system.
Description
Technical Field
The invention relates to the technical field of optical imaging, in particular to a microscope system, an adjusting method and a sequencing system thereof.
Background
In a high-resolution or ultra-high-resolution microscope imaging system, the high-resolution or ultra-high-resolution microscope imaging system is often used in cooperation with a high-power objective lens, and because the depth of field range of the high-power objective lens is very small, if the parallelism between an object carrying surface of an object carrying platform of the microscope system and a plane on which the objective lens is mounted is poor, a clear image cannot be obtained by the high-power objective lens.
Therefore, there is still a need for an improvement in the construction of autocollimators and/or in how to effectively and easily adjust the parallelism of the object-carrying surface of the object-carrying platform of a microscope system with respect to the plane in which the objective is mounted.
Disclosure of Invention
In view of the above, the present invention provides a microscope system, an adjusting method thereof and a sequencing system.
In a tuning method of a microscope system according to an embodiment of the present invention, the microscope system includes a light source, an object stage, and an objective lens mounting member disposed opposite to the object stage, the object stage includes an object carrying surface, and the objective lens mounting member includes an objective lens mounting surface opposite to the object carrying surface, and the tuning method includes:
transmitting light passing through a reticle having at least one identification pattern to the objective lens mounting surface with a light source;
acquiring a first image of the identification pattern formed by the light reflected by the objective lens mounting surface;
transmitting light passing through the reticle to the object carrying surface by using the light source;
acquiring a second image of the identification pattern formed by the light reflected by the object carrying surface; and
and adjusting the object carrying platform to enable the second image of the identification pattern to be superposed with the first image of the identification pattern, so that the object carrying surface is parallel to the objective lens mounting surface.
The microscope system of the embodiment of the invention is obtained by the adjusting method.
The test system of the embodiment of the invention comprises the microscope system.
In the adjusting method of the microscope system and the microscope system, whether the object carrying surface is parallel to the objective lens mounting surface or not is detected by light rays of the light source passing through the reticle, the process is simple and convenient, the efficiency of detecting whether the object carrying surface is parallel to the objective lens mounting surface or not is improved, and the adjusting precision is high. In addition, the object carrying platform is adjusted when the object carrying surface is not parallel to the objective lens mounting surface, so that the object carrying surface is parallel to the objective lens mounting surface, and the adjusting process is simple.
Additional aspects and advantages of the invention will be set forth in part in the description which follows and, in part, will be obvious from the description, or may be learned by practice of the invention.
Drawings
The above and/or additional aspects and advantages of the present invention will become apparent and readily appreciated from the following description of the embodiments, taken in conjunction with the accompanying drawings of which:
FIG. 1 is a schematic structural diagram of a microscope system according to an embodiment of the present invention;
FIG. 2 is another schematic structural view of a microscope system according to an embodiment of the present invention;
FIG. 3 is a schematic view of yet another configuration of a microscope system according to an embodiment of the present invention;
FIG. 4 is a schematic view of a scene of a microscope system according to an embodiment of the invention;
FIG. 5 is a partial perspective view of a microscope system according to an embodiment of the present invention;
FIG. 6 is another perspective view of a portion of a microscope system according to an embodiment of the present invention;
FIG. 7 is a schematic partial cross-sectional view of a microscope system according to an embodiment of the invention;
FIG. 8 is an enlarged schematic view of a portion V of the microscope system of FIG. 7;
FIG. 9 is a schematic view of the connection of the support member and the adjustment member of an embodiment of the present invention;
FIG. 10 is another schematic cross-sectional view of a portion of a microscope system according to an embodiment of the invention;
FIG. 11 is an enlarged schematic view of a VI portion of the microscope system of FIG. 10;
FIG. 12 is a schematic view of the connection of a second adjustment structure and a carrier module of an embodiment of the present invention;
FIG. 13 is another schematic view of a second adjustment structure and a carrier module according to an embodiment of the present invention;
FIG. 14 is a plan view of a second adjustment structure and carrier module of an embodiment of the invention;
FIG. 15 is a schematic perspective view of a sequencing system according to an embodiment of the invention;
FIG. 16 is another perspective view of a sequencing system according to an embodiment of the present invention.
Fig. 17 is a flowchart illustrating a method for adjusting a microscope system according to an embodiment of the present invention.
FIG. 18 is a schematic illustration of the position of the reactor surface in an embodiment of the invention.
Description of the main element symbols:
Detailed Description
Reference will now be made in detail to embodiments of the present invention, examples of which are illustrated in the accompanying drawings, wherein like reference numerals refer to the same or similar elements or elements having the same or similar functions throughout. The embodiments described below with reference to the accompanying drawings are illustrative only for the purpose of explaining the present invention, and are not to be construed as limiting the present invention.
In the present invention, unless otherwise expressly specified or limited, the first feature "on" or "under" the second feature may comprise direct contact of the first and second features, or may comprise non-direct contact of the first and second features. Also, the first feature being "on," "above" and "over" the second feature includes the first feature being directly on and obliquely above the second feature, or merely indicating that the first feature is at a higher level than the second feature. A first feature being "under," "below," and "beneath" a second feature includes the first feature being directly under and obliquely below the second feature, or simply meaning that the first feature is at a lesser elevation than the second feature.
In the description of the present invention, it should be noted that, unless otherwise explicitly specified or limited, the terms "mounted," "connected," and "connected" are to be construed broadly, e.g., as meaning either a fixed connection, a removable connection, or an integral connection; may be mechanically, electrically or may be in communication with each other; either directly or indirectly through intervening media, either internally or in any other relationship. The specific meanings of the above terms in the present invention can be understood according to specific situations by those of ordinary skill in the art.
Referring to fig. 1, a microscope system 100 according to an embodiment of the present invention includes a frame 10, a stage 20, an objective lens mounting member 30, a light source assembly 40, a camera 50, a processor 60, a focusing lens 70, a dichroic mirror 80, and a first adjusting mechanism 90. Both the stage 20 and the objective lens mount 30 are mounted on the frame 10. The processor 60 is coupled to the camera 50 and the first adjustment mechanism 90 is coupled to the carrier platform 20.
In particular, the gantry 10 is a carrier of the microscope system 100, which is used to carry components of the microscope system 100. For example, both the stage 20 and the objective lens stage are mounted on the frame 10. In the illustration of fig. 1, the frame 10 is L-shaped, and it is understood that in other embodiments, the structure and shape of the frame 10 may be designed to be any shape according to requirements, for example, the frame 10 has an irregular structure. The frame 10 may be formed by assembling a plurality of parts, for example, the frame 10 may be formed by fixedly connecting a plurality of parts by fasteners such as screws.
The stage 20 is used to load an object such as a reactor, for example, a slide on which a tissue section to be observed is placed. Stage 20 may be moved in different directions to adjust the position of the sample on stage 20. For example, the stage 20 may move vertically, or may move back and forth and left and right in the horizontal direction. The carrier platform 20 may be generally circular, square or other irregular shape. The carrier platform 20 comprises a carrier surface 21. The carrier surface 21 is used for carrying objects such as a reactor, for example, a sample can be clamped by a clamp after being carried on the carrier surface 21 to prevent the sample from moving on the carrier surface 21.
The objective lens mount 30 is disposed opposite to the stage 20 and is optically transparent, and the objective lens mount 30 is used to mount an objective lens. For example, the objective lens mount 30 is provided with a screw interface or a snap interface for mounting the objective lens. It is understood that the objective lens includes a plurality of lenses, and the plurality of lens combinations can obtain the corresponding imaging multiples. For example, the objective lens has a magnification of 10 times or the like, so that the microscopic structure of the sample can be observed by the microscope system 100. The objective lens may serve as the above focusing lens 70.
The objective lens mount 30 includes an objective lens mounting surface 31, and the objective lens mounting surface 31 is opposed to the object mount surface 21. It should be noted that "opposite" as referred to herein may mean that the two planes are oriented substantially oppositely, or that the two planes are oriented substantially the same. For example, the objective lens mounting surface 31 faces downward, and the object mounting surface 21 faces upward. For another example, the objective lens mounting surface 31 and the object mount surface 21 are both oriented upward. In the present embodiment, the objective lens attachment surface 31 is a surface facing downward of the objective lens attachment device 30.
It should be noted that the objective lens mounting member 30 is light-transmissive, and the objective lens mounting member 30 may be made of a light-transmissive material, for example, the material of the objective lens mounting member 30 is glass or a resin material; the objective lens mounting member 30 may be formed with a through hole through which light passes so as to transmit light.
The light source assembly 40 is used for emitting light to the stage 20 and the objective lens mounting member 30. Specifically, the light source assembly 40 includes a light source 41, a collimating lens 42, and a reticle 43, and the light source 41, the collimating lens 42, and the reticle 43 are sequentially disposed along a conduction path of the light source 41. It is understood that the light source 41 generates light having a transmission path, and the projection path of the light source 41 may be a straight line type or a broken line type. The light source 41, the collimator lens 42, and the reticle 43 are arranged in this order along the same straight line.
The light source 41 is used for emitting light, and in the present embodiment, the light source 41 is used for emitting light passing through the reticle 43 to the objective lens mounting surface 31 or the object mounting surface 21.
The light emitted by the light source 41 is, for example, visible light. The light source 41 is, for example, an LED lamp, so that the light source 41 has the advantages of low energy consumption, long service life and the like. The power of the light source 41 can be specifically set according to actual conditions, for example, when the microscope system 100 is located in a dark environment, the power of the light source 41 is higher to emit light with higher brightness, so that the microscope system 100 can obtain a clearer sample image.
The light source 41 is, for example, a point light source, that is, a light beam emitted from the light source 41 so as to be dispersed around the center thereof.
The collimator lens 42 is located in front of the light source 41, or the light source 41 is used to project light into the collimator lens 42. The collimating lens 42 is used for collimating the light emitted from the light source 41, or the collimating lens 42 is used for converging the divergent light emitted from the light source 41 to form parallel light, so that the light emitted from the collimating lens 42 is a parallel light beam. It is understood that the collimating lens 42 includes a collimating lens.
It should be noted that in other embodiments, when the light emitted from the light source 41 is parallel light, the collimating lens 42 may be omitted.
The reticle 43 has at least one logo so that light transmitted through the reticle 43 carries logo information. The reticle 43 is made of a light-transmitting material, and the material of the reticle 43 is glass, for example. The identification pattern on the reticle 43 may be formed by a hollowing process. The marking pattern on the reticle 43 is, for example, a pattern of a graduated cross-hair line or two concentric circles.
Referring to fig. 4, the camera 50 is used for receiving a first image 51 of the identification pattern formed by the light reflected by the objective lens mounting surface 31 and a second image 52 of the identification pattern formed by the light reflected by the object carrying surface 21. Alternatively, the camera 50 is used to receive a first image 51 and a second image 52. In the present embodiment, the first image 51 and the second image 52 are both cross-hatched images.
The light reflected by the objective lens mounting surface 31 and the object mount surface 21 can be obtained by mounting a light reflecting element, for example, a light reflected by the first optical element or the second optical element described later, on the objective lens mounting surface 31 or the object mount surface 21.
In one example, the size of the camera electronic sensor is 6.5um, the magnification of the imaging optical path is 60, the minimum size that the microscope system 100 can resolve is 0.1um, and when the first image and the second image are not more than 20 pixels apart on the same image plane, it is determined that the second image and the first image coincide.
Specifically, if necessary, the first image 51 and the second image may be identified by image processing methods (e.g., filtering, interpolation, etc.), and then whether the distance between the two images is greater than 20 pixels may be calculated, where the pixels are image pixels of the image acquired by the camera 50. Since both the first image 51 and the second image 52 can be acquired by the same camera, in case the first image 51 and the second image 52 are acquired by the camera 50, the imaging planes of the two images on the camera 50 are the same image plane, e.g. the surface of the image sensor of the camera 50. The distance between the images is used to determine whether the two images coincide, the distance threshold is set according to the resolution (or magnification) of the camera 50, the field of view of the objective lens, the depth of field, the shape of the image, etc., and one skilled in the art can set the distance threshold based on the resolution of the camera 50 and the shape of the image, and the requirements for accuracy in combination with different optical detection applications. For example, the shape of the image is a point or a circle, and the distance between the center points of two points or two circles can be used as the distance between two images; for another example, the shape of the image is a straight line, and the distance between the centers of two straight lines can be used as the distance between the two images; for another example, the shape of the image is a cross, and the distance between the two images may be the distance between the intersections of two crosses.
The camera 50 is, for example, a camera, such as an RGB camera, to acquire a color image.
Referring to fig. 2, in the present embodiment, the microscope system 100 includes a first optical element 102 disposed on the objective lens mounting surface 31, and the first optical element 102 is used for reflecting light reaching the objective lens mounting surface 31. In this manner, the camera 50 can capture a first image formed by the light reflected by the first optical element 102.
Referring to fig. 3, in the present embodiment, the microscope system 100 includes a second optical element 104 disposed on the object plane 21, and the second optical element 104 is used for reflecting light reaching the object plane 21. In this manner, the camera 50 may capture a second image formed by light reflected by the second optical element 104.
It is understood that the first optical element 102 has a surface parallel to the objective lens mounting surface 31, and the second optical element 104 has a surface parallel to the object mount surface 21.
The first and second images are acquired separately, and in one example, the objective lens mount 30 is positioned to mount the objective lens when the second image 52 is acquired.
In some embodiments, the first optical element 102 and/or the second optical element 104 are materials with light reflection capability and have a better flatness and parallelism to enable parallel light reaching the surface to be reflected and still be parallel light; may be selected from at least one of glass, flat mirrors, and parallel flats.
Specifically, the first optical element 102 and the second optical element 104 may be selected from the same or different types of optical elements. For example, the first optical element 102 is a plane mirror, the second optical element 104 is a parallel plate, and for example, both the first optical element 102 and the second optical element 104 are mirrors or parallel plates.
The processor 60 is connected to the camera 50, and the processor 60 can be used to control the camera 50 to image.
The focusing lens 70 is disposed between the camera 50 and the objective lens mount 30, the focusing lens 70 serves to condense the parallel light reflected by the objective lens mounting surface 31 or the object carrying surface 21, and the camera 50 serves to receive the light condensed by the focusing lens 70. In one example, the objective lens mount 30 is positioned to mount an objective lens, referred to as a focusing lens 70, when a second image is acquired.
The dichroic mirror 80 is disposed between the reticle 43 and the stage 20, the dichroic mirror 80 is configured to reflect light emitted from the light source 41 through the reticle 43 to reflect the light to the objective lens mounting surface 31 or the object mounting surface 21, and the dichroic mirror 80 is further configured to transmit the light reflected by the objective lens mounting surface 31 or the object mounting surface 21.
In one example, the dichroic mirror 80 is disposed to be inclined at 45 degrees with respect to the horizontal direction, and the angles at which the light emitted from the light source 41 is reflected by the dichroic mirror 80 and then turned are all 90 degrees, or the angle between the light before being reflected by the dichroic mirror 80 is 90 degrees.
In some embodiments, camera 50 is used to acquire a first image formed by light reflected by objective lens mounting surface 31 and transmitted through dichroic mirror 80;
the camera 50 can also be used to acquire a second image formed by the light reflected by the object bearing surface 21 and transmitted through the dichroic mirror 80. Dichroic mirrors, also known as dichroic mirrors, have the property of being almost completely transmissive for light of certain wavelengths and almost completely reflective for light of other wavelengths. The inventors provided a specific dichroic mirror 80 in the reflection optical path to reflect the incident light and not to act on the reflected light (the reflected light is transmitted).
Referring to fig. 4, a first adjusting structure 90 is connected to the object stage 20, and the first adjusting structure 90 is used for adjusting the object stage 20 to make the first image 51 of the identification pattern coincide with the second image 52 of the identification pattern, so that the object carrying surface 21 is parallel to the objective lens mounting surface 31. In this way, when the first image 51 of the first identification pattern and the second image 52 of the identification pattern coincide, it is indicated that the object carrying surface 21 and the objective lens mounting surface 31 are parallel, which is advantageous for the objective lens to obtain a clear image. Therefore, when the object carrying surface 21 is not parallel to the objective lens mounting surface 31, the object carrying platform 20 is adjusted to make the object carrying surface parallel to the objective lens mounting surface, and the adjustment process is simple.
The first adjusting mechanism 90 is, for example, a power transmission device composed of a gear mechanism, a motor, a rail mechanism, etc., which can drive the carrier platform 20 to adjust the position of the carrier platform 20.
Specifically, in this embodiment, referring to fig. 5, the first adjusting structure 90 includes a first adjusting member 116 and a supporting member 118.
Referring to fig. 6-9, the supporting member 118 is provided with an inclined surface 120, the loading platform 20 is disposed on the inclined surface 120, the first adjusting member 116 is connected to the supporting member 118, and the first adjusting member 116 is adjusted to drive the supporting member 118 to move so as to change the position of the loading platform 20 on the inclined surface 120, so as to adjust the loading platform 20, and further, the second image 52 is overlapped with the first image 51. In this way, the second image 52 is overlapped with the first image 51 by adjusting the position of the stage 20 on the inclined plane 120, and the adjustment method is simple and easy to implement. The object carrying surface 21 may be an upper surface of the object carrying platform 20.
Specifically, the first adjusting structure 90 includes a base plate 122, the first adjusting member 116 and the supporting member 118 are disposed on the base plate 122, the stage 20 is located on the base plate 122, and the pitch angle of the object carrying surface 21 relative to the base plate 122 is adjusted when the first adjusting member 116 is adjusted to drive the supporting member 118.
In one example, the first adjusting member 116 is a screw, the first adjusting member 116 is in threaded connection with the supporting member 118, the base plate 122 defines a limiting groove 124, the supporting member 118 is disposed in the limiting groove 124, and the limiting groove 124 is used for limiting the rotation of the supporting member 118 relative to the first adjusting member 116. When the first adjusting member 116 rotates, the supporting member 118 can only move linearly back and forth along the length direction of the first adjusting member 116 due to the limiting groove 124, so that the position of the carrier platform 20 on the inclined surface 120 is adjusted.
In this embodiment, the first adjustment structure 90 includes a spacer 126, a first elastic member 128, a connection screw 130, and a mating component 132, the connection screw 130 includes a head portion 134 and a post portion 136, and the head portion 134 includes a flange protruding from the post portion 136. The stage platform 20 has a first connecting hole 138, and the first connecting hole 138 is stepped. The attachment screw 130 extends through the first attachment opening 138, with the head portion 134 and a portion of the post portion 136 received in a larger section of the first attachment opening 138 and another portion of the post portion 136 extending through a smaller section of the first attachment opening 138 and attached to the base plate 122. The first elastic member 128 is received between the head portion 134 and the bottom surface of the larger section of the first connection through hole 138, so that the substrate 122 and the stage 20 can be elastically connected.
The pad 126 is sandwiched between the mating component 132 and the ramp 120. The support 117 is formed with a through hole 140 passing through the inclined surface 120 and penetrating the support, and the post 136 of the connection screw 130 is formed with the through hole 140. Referring to FIG. 8, the space between the left and right sidewalls of the through hole 140 and the post 136 of the connection screw 130 is large enough so that the connection screw 130 does not block the desired displacement of the support member 118 when moving back and forth along the axis of the first adjustment member 116.
To facilitate the pitch angle of the carrier 20, the engaging component 132 includes a first engaging element 142 and a second engaging element 144, the first engaging element 142 is disposed on the bottom surface of the carrier 20, and the second engaging element 144 is disposed in a recess on the top surface of the pad 126. The first mating member 142 includes a first mating surface 146 having a circular arc shape, the second mating member 144 includes a second mating surface 148 having a circular arc shape, and the first mating surface 146 and the second mating surface 148 are rotatably connected.
In fig. 5, the first adjusting member 116, the supporting member 118, the pad 126, the first elastic member 128, the connecting screw 130 and the matching component 132 are disposed on both left and right sides of the carrier platform 20, so as to achieve more accurate pitch adjustment. It is understood that in other embodiments, the first adjustment member 116 and the support member 118 may be provided on a single side of the carrier platform 20. If resilient support and smoother angular adjustment are desired, a spacer 126, a first resilient member 128, a coupling screw 130 and a mating member 132 may be added.
Further, referring to fig. 10 and fig. 11, the first adjusting structure 90 includes a connecting member 150, a first fitting 152 and a second fitting 154, the connecting member 150 connects the carrier platform 20 and the base plate 122, the first fitting 152 and the fourth fitting 154 are relatively rotatably connected and located between the carrier platform 20 and the base plate 122, the first fitting 152 is disposed on the carrier platform 20, the second fitting 154 is disposed on the base plate 122, so that the first fitting 152 and the second fitting 154 are relatively rotated to adjust the pitch angle of the carrier surface 21 relative to the base plate 122.
Specifically, in fig. 5, the first adjusting member 116 and the supporting member 118 are located at the left side and/or the right side of the stage 20 closer to the front side of the microscope system 100, and the connecting member 150, the first fitting member 152 and the second fitting member 154 are located at the rear side of the stage 20, so that an adjusting scheme that the pitch angle can be adjusted at the front side and the rear side is used as a rotation point is formed.
The connecting member 150 may be a screw, the stage 20 is provided with a second connecting through hole 154, the second connecting through hole 155 is stepped, the connecting member 150 includes a head portion and a pillar portion, and the head portion includes a flange protruding from the pillar portion. The connecting member 150 is inserted into the second connecting through hole 155, a part of the head and the column of the connecting member 150 is received in the larger section of the second connecting through hole 155, and another part of the column of the connecting member 150 passes through the smaller section of the second connecting through hole 155 and is connected to the carrier platform 20. A second elastic member 156 is received between the head of the connecting member 150 and the bottom surface of the larger section of the second connecting through hole 155, so that the substrate 122 and the stage 20 can be elastically connected.
To facilitate the tilting angle of the carrier platform 20, the first fitting member 152 includes a third fitting surface 158 having a circular arc shape, the second fitting member 154 includes a fourth fitting surface 160 having a circular arc shape, and the third fitting surface 158 and the fourth fitting surface 160 are rotatably connected.
In some embodiments, referring to fig. 5, first adjustment mechanism 90 includes a securing assembly 162, securing assembly 162 being configured to secure stage 20 after first image 51 and second image 52 are registered. Specifically, the fixing assembly 162 includes a fixing plate 164 and a fixing member 166, the fixing plate 164 is L-shaped, one side plate of the fixing plate 164 is connected to the upper surface of the base plate 122, and the other side plate is connected to the side surface of the carrier platform 20. Fasteners 166 fixedly couple mounting plate 164 to carrier platform 20 and base plate 122. The fixing member 166 may be a screw.
In fig. 5, the fixing members 162 are disposed on both left and right sides of the carrier platform 20. This ensures that the carrier platform 20 is stable.
The microscope system 100 comprises a carrier module 107 and a second conditioning structure 108, the carrier module 107 being adapted to carry the reactor 200. The carrier module 107 is located on the second adjusting structure 108. The second adjustment structure 108 is used to enable the reactor 203 and the object carrying surface 21 to satisfy a predetermined positional relationship.
The second adjusting structure 108 includes an adjusting plate 168 and a plurality of second adjusting members 170, the carrying module 107 includes a base 172 disposed on the adjusting plate 168, the plurality of second adjusting members 170 are disposed at intervals and movably connected to the base 172 and the adjusting plate 168, and the second adjusting members 170 are adjusted to drive the adjusting plate 168 to enable the reactor 203 and the object carrying surface 21 to satisfy a predetermined position relationship. In this way, the plane 202 of the reactor and the object carrying surface 21 meet the preset position relationship through multi-point adjustment.
Specifically, microscope system 100 includes a movable stage 174, wherein movable stage 174 is disposed on stage 20, and second adjustment structure 108 is disposed on movable stage 172, and movable stage 174 is capable of moving second adjustment structure 108 and reactor 200 in a direction perpendicular to optical axis OP of the lens.
In this embodiment, please refer to fig. 12 to 14, the second adjusting member 170 includes two adjusting screws 176, a third elastic member and a matching component. The adjusting screw 176 and the third elastic member can realize the elastic connection between the adjusting plate 168 and the base 172, and the elastic connection between the base plate 122 and the carrier platform 20 can be referred to, and the base 172 can be more smoothly adjusted by cooperating with the assembly, and the carrier platform 20 can be referred to more smoothly adjusted. Two adjustment screws 176 are located on the outside of the mating assembly. An adjusting screw 176 connects the adjusting plate 168 and the base 172, and the distance between the adjusting plate 168 and the base 172 is adjusted by the adjusting screw 176. Specifically, for each second adjusting member 170, the distance between the adjusting plate 168 and the base 172 is adjusted by screwing in and out two adjusting screws 176, so that the plane 202 of the reactor and the object carrying surface 21 can satisfy the preset positional relationship by the adjustment of the plurality of second adjusting members 170.
In the example of fig. 14, the number of the second adjusters 170 is three, and three second adjusters 170 are distributed in an isosceles triangle.
Specifically, referring to fig. 14, two second adjusting members 170 are respectively disposed on the left and right sides of the adjusting plate 168 at positions closer to the front side of the microscope system 100, and another second adjusting member 170 is disposed on the rear side of the second adjusting plate. The connecting line of the two second adjusting members 170 is the bottom side of an isosceles triangle, and the two connecting lines of the two second adjusting members 170 and the other second adjusting member 170 are the two waists of the isosceles triangle.
In some embodiments, the base 172 is provided with a receiving groove 178 for receiving the reactor 200, and the receiving groove 178 is provided with a positioning structure 180 for positioning the reactor 200. Thus, the positioning structures 180 are respectively arranged in the accommodating groove 178, so that when the reactor 200 is accommodated in the accommodating groove 178, the positioning structures 180 can pre-position the reactor 200 well, and the establishment of a flow path is ensured.
Specifically, the positioning structure 180 includes three positioning pillars 182, and the three positioning pillars 182 are distributed on two adjacent sides of the accommodating groove 178. In this manner, a three-point alignment of the reactor 200 can be achieved.
In fig. 13, the planar shape of the receiving groove 178 is substantially square, two positioning pillars 182 are located at the upper side of the receiving groove 178, and one positioning pillar 182 is located at the right side of the receiving groove 178. Two positioning posts 182 may be disposed in a direction parallel to the channel of the reactor 200. Positioning posts 182 may be positioning pins.
In addition, the carrying module 107 includes a side pushing mechanism 184 at the joint of the left side and the lower side, and the side pushing mechanism 184 is telescopically disposed in the accommodating groove 178 and is used for ensuring that the reactor 200 is tightly attached to the positioning structure 180.
Thus, the side pushing mechanism 184 can be matched with the waist hole of the reactor, so that the over-constraint of the accommodating of the reactor 200 in the accommodating groove 178 is avoided, and meanwhile, the side pushing mechanism 184 ensures that the reactor 200 is tightly attached to the positioning structure 180, so as to realize positioning and fixing.
The side pushing mechanism 184 includes a side pushing element 186 and a fourth elastic element (not shown), the fourth elastic element is disposed in the base 172, the side pushing element 186 is connected to the fourth elastic element and partially protrudes into the accommodating groove 178, so that when the reactor 200 is accommodated in the accommodating groove 178, the fourth elastic element can apply an elastic force to the reactor 200 through the side pushing element 186, so that the reactor 200 is tightly attached to the positioning pillars 182 on the upper side and the right side.
Referring to fig. 15 and 16, a sequencing system 300 according to an embodiment of the present invention includes an optical imaging system 400 according to any of the above embodiments. The sequencing system 300 can implement the method of any of the embodiments described above.
Specifically, the sequencing system 300 includes a reagent cartridge 188 for storing reagents, and a rotary valve, a three-way valve, a carrying module 107 and a power device 190 are arranged in the flow direction of a liquid flowing out from the reagent cartridge 188. The carrier module 107 houses the reactor 200, and the channels of the reactor 200 are connected to the flow path of the sequencing system 300. Different reagents can be introduced into the flow path through the three-way valve by the rotary valve to perform different reactions within the channels of the reactor 200, including but not limited to extension, excision, capping, imaging, washing, and the like.
The power unit 190 may employ a pump to power the liquid in the flow path.
Referring to FIG. 5, the microscope system 100 is mounted to the sequencing system 100 by 4 legs 192.
In other embodiments, the sequencing system 300 may omit a three-way valve, which may direct different reagents into the channels of the reactor 200.
Referring to fig. 17, the present invention further provides a method for adjusting the microscope system 100, which includes the following steps:
s10, emitting light passing through a reticle 43 to an objective lens mounting surface 31 by using a light source 41, wherein the reticle 43 is provided with at least one identification pattern;
s20, acquiring a first image 51 of an identification pattern formed by light reflected by the objective lens mounting surface 31;
s30, emitting light passing through the reticle 43 to the object carrying surface 21 by using the light source 41;
s40, acquiring a second image 52 of the identification pattern formed by the light reflected by the object carrying surface 21;
s50, adjusting the carrying platform 20 to enable the second image 21 of the identification pattern to be superposed with the first image 51 of the identification pattern, so that the carrying surface 21 is parallel to the objective lens mounting surface 31.
In the adjusting method of the microscope system 100 and the microscope system 100, the light of the light source 41 passing through the reticle 43 is used for detecting whether the object carrying surface 21 and the objective lens mounting surface 31 are parallel, the process is simple, the efficiency of detecting whether the object carrying surface 21 and the objective lens mounting surface 31 are parallel is improved, and the adjusting precision is high. In addition, when the object carrying surface 21 is not parallel to the objective lens mounting surface 31, the object carrying platform 20 is adjusted to enable the object carrying surface 21 to be parallel to the objective lens mounting surface 31, and the adjusting process is simple.
In some embodiments, a tuning method comprises:
the carrier platform 20 is secured with the securing assembly 162 when the second image 52 and the first image 51 are coincident.
In this way, after the object platform 20 is adjusted, the fixing assembly 160 can be used to fix the position of the first adjusting plate 114, so as to ensure that the position of the object carrying surface 21 is unchanged.
In some embodiments, a tuning method comprises:
mounting the carrying module 107 carrying the reactor 200 and the second adjusting structure 108 on the carrying surface 21, wherein the carrying module 107 is positioned on the second adjusting structure 108;
the second adjustment structure 108 is adjusted so that the surface 203 of the reactor 200 and the loading surface 21 satisfy a predetermined positional relationship.
In this way, the surface 203 of the reactor 200 can be adjusted to the carrier surface 21 to satisfy the predetermined positional relationship, so that the positional relationship between the plane of the reactor 200 and the camera 50 can be adjusted to a desired positional relationship, and the requirements of sequencing are satisfied, including the steps of optically detecting the specific position of the reactor 200 by the camera 50 during the sequencing process and optically detecting a plurality of specific positions of the reactor 200 during the dynamic process.
The optical imaging system 400 can be used in the sequencing system 300, and the camera 50 needs to take a picture of the reactor 200 during the sequencing process. Therefore, the positional relationship between the camera 50 and the reactor 200 needs to be adjusted to a desired positional relationship, for example, the camera 50 includes an optical axis, and the adjustment method aims to make the optical axis satisfy a perpendicular relationship with the surface 203 of the reactor 200, and to keep the optical axis constantly in a perpendicular relationship with the surface 203 of the reactor 200 during the relative movement of the reactor 200 and the camera 50.
In the embodiment of the present invention, the position of the reactor 200 refers to a surface region of the reactor 200 as the position of the reactor 200 when the surface region is set below the camera 50 and imaged by the camera 50.
The purpose of adjusting the first adjusting structure 90 to make the second image 52 coincide with the first image 51 is to make the optical axes of the object carrying surface 21 and the camera 50 satisfy a preset positional relationship, for example, when the object carrying surface 21 is the upper surface of the first adjusting structure 90, the optical axes of the object carrying surface 21 and the camera 50 may satisfy a vertical positional relationship; when the object carrying surface 21 is a side surface perpendicular to the upper surface of the first adjusting structure 90, the object carrying surface 21 and the optical axis of the camera 50 can satisfy a parallel position relationship.
The first adjusting structure 90 is adjusted to make the second image 52 coincide with the first image 51, in practice, after the camera 50 acquires the first image 51, the objective lens mounting surface 31 can be fixed to make the first image 51 fixed, then the first adjusting structure 90 is used to adjust the position relationship between the second image 52 and the first image 51 until a certain second image 52 coincides with the first image 51, and then the first adjusting structure 90 is fixed so that the first adjusting structure 90 is in a state/position capable of obtaining the second image 52 coinciding with the first image 51.
In some embodiments, adjusting the second adjustment structure 108 to allow the surface 203 of the reactor 200 and the carrier surface 21 to satisfy a predetermined positional relationship comprises:
the second adjusting member 170 is adjusted such that the absolute value of the difference between the first distance and the second distance is less than the first set value, and the absolute value of the difference between the third distance and the fourth distance is less than the second set value,
the first distance is the distance from the camera 50 at the first location 11 on the surface of the reactor 200,
the second distance is the distance from the camera 50 to the second location 12 on the surface of the reactor 200,
the third distance is the distance from the camera 50 to the third location 13 on the surface of the reactor 200,
the fourth distance is the distance from the camera 50 at the fourth location 14 on the surface of the reactor 200,
the line connecting the first position 11 and the second position 12 is non-parallel to the line connecting the third position 13 and the fourth position 14.
Specifically, referring to fig. 18, the planar shape of the reactor 200 is rectangular, the first position 11 is located as close to the left side of the surface of the reactor 200 as possible, the second position 12 is located as close to the right side of the surface of the reactor 200 as possible, and the connection line 15 is parallel to the length direction of the reactor 200. The third position 13 is located as close as possible to the upper side of the surface of the reactor 200, the fourth position 14 is located as close as possible to the lower side of the surface of the reactor 200, and the line 16 is parallel to the width direction of the reactor. That is, in the example of fig. 18, the line 15 is perpendicular to the line 16.
The distance between the position of the surface of the reactor 200 and the camera 50 refers to the distance between the position of the surface of the reactor 200 and the camera 50 in the direction in which the optical axis of the camera 50 extends.
Specifically, adjusting the second adjusting structure 108 to make the surface 203 of the reactor 200 and the object carrying surface 21 satisfy a preset positional relationship includes:
adjusting a portion of the second adjusting member 170 so that a distance between the first position 11 and the second position 12 on the surface of the reactor 200 in the direction of reflected light is smaller than a first set value;
and fixing a part of the second adjusting member 170, and adjusting another part of the second adjusting member 170 so that a distance between the third position 13 and the fourth position 14 on the surface of the reactor 200 in the direction of reflected light is smaller than a second set value.
In fig. 18, the longitudinal direction (transverse direction) of the reactor 200 is defined as the x-direction, and the width direction (longitudinal direction) of the reactor 200 is defined as the y-direction.
During adjustment, 1) the second adjusting member 170 fixed to the rear side of the adjusting plate 168 is first used, and the two second adjusting members 170 in the x direction are adjusted so that the x direction of the reactor 200 is parallel to the x direction of the object carrying surface 21 after the second-stage adjustment. Specifically, the camera 50 focuses on the first position 11 and the second position 12, and the values obtained after focusing are respectively used as the first distance Zx1 and the second distance Zx2, | Zx1-Zx2| is smaller than the first set value, it is determined that the x direction of the reactor 200 is parallel to the x direction of the object carrying surface 21 after the secondary adjustment. In one example, the first setting is 3 microns.
2) Fixing two second adjusting pieces 170 in the x direction in 1), adjusting the second adjusting pieces 170 at the rear side of the adjusting plate 168 to make the surface 203 of the reactor 200 parallel to the object carrying surface 21, specifically, focusing the third position 13 and the fourth position 14 by the imaging assembly 110, respectively, and determining that the surface of the reactor 200 is parallel to the object carrying surface 21 after the second-stage adjustment when the values obtained after focusing are respectively taken as the third distance Zy1 and the fourth distance Zy2, | Zy1-Zy2| is smaller than the second set value. In one example, the second setting is 5 microns. It is understood that in other examples, the first set point and the second set point may take other values, and the first set point and the second set point may be the same or different.
In the description herein, references to the description of the terms "one embodiment," "certain embodiments," "an illustrative embodiment," "an example," "a specific example," or "some examples," etc., mean that a particular feature, structure, material, or characteristic described in connection with the embodiment or example is included in at least one embodiment or example of the invention. In this specification, schematic representations of the above terms do not necessarily refer to the same embodiment or example. Furthermore, the particular features, structures, materials, or characteristics described may be combined in any suitable manner in any one or more embodiments or examples.
While embodiments of the present invention have been shown and described, it will be understood by those of ordinary skill in the art that: various changes, modifications, substitutions and alterations can be made to the embodiments without departing from the principles and spirit of the invention, the scope of which is defined by the claims and their equivalents.
Claims (15)
1. A method of tuning a microscope system, the microscope system including a light source, a stage including an object-carrying surface, and an objective lens mount disposed opposite the stage and including an objective lens mounting surface opposite the object-carrying surface, the method comprising the steps of:
transmitting light passing through a reticle having at least one identification pattern to the objective lens mounting surface with a light source;
acquiring a first image of the identification pattern formed by the light reflected by the objective lens mounting surface;
transmitting light passing through the reticle to the object carrying surface by using the light source;
acquiring a second image of the identification pattern formed by the light reflected by the object carrying surface; and
adjusting the object carrying platform to enable the second image of the identification pattern to be superposed with the first image of the identification pattern, so that the object carrying surface is parallel to the objective lens mounting surface;
the microscope system comprises a bearing module and a second adjusting structure, wherein the bearing module is used for bearing a reactor, and the adjusting method comprises the following steps:
mounting a carrier module carrying the reactor and the second conditioning structure on the carrier surface, the carrier module being located on the second conditioning structure,
adjusting the second adjusting structure to enable the surface of the reactor and the object carrying surface to meet a preset position relation;
the second adjustment structure and the reactor are moved in a direction perpendicular to the optical axis of the lens.
2. The method of tuning a microscope system according to claim 1, wherein the microscope system comprises a dichroic mirror disposed between the reticle and the stage.
3. The method of adjusting a microscope system according to claim 1, wherein the objective lens mounting surface is provided with a first optical element for reflecting the light reaching the objective lens mounting surface; or
The object carrying surface is provided with a second optical element for reflecting light reaching the object carrying surface.
4. The method of tuning a microscope system according to claim 3, wherein the first optical element and/or the second optical element is selected from at least one of glass, a flat mirror, a mirror, and a parallel plate.
5. The method for adjusting a microscope system according to claim 1, wherein the microscope system includes a camera for receiving the first image and the second image, and when the distance between the first image and the second image on the same image plane is not more than 20 pixels, it is determined that the second image and the first image coincide.
6. The method of claim 1, wherein the microscope system comprises a first adjustment mechanism coupled to the stage, the first adjustment mechanism configured to adjust the stage.
7. The method of claim 6, wherein the first adjusting structure comprises a first adjusting member and a supporting member, the supporting member has an inclined surface, the stage is disposed on the inclined surface, the first adjusting member is connected to the supporting member, and the first adjusting member is adjusted to move the supporting member so as to change the position of the stage on the inclined surface, thereby adjusting the stage.
8. The method of claim 7, wherein the first adjusting structure comprises a base plate, the first adjusting member and the supporting member are disposed on the base plate, the stage is disposed on the base plate, and the first adjusting member is adjusted to adjust the pitch angle of the stage surface relative to the base plate when the supporting member is moved.
9. The method of claim 8, wherein the first adjustment structure comprises a connector connecting the stage and the base plate, a first fitting and a second fitting, the first fitting and the second fitting are rotatably connected and located between the stage and the base plate, the first fitting is disposed on the stage, the second fitting is disposed on the base plate, and the first fitting and the second fitting are rotated relative to each other to adjust the pitch angle of the stage relative to the base plate.
10. The method of tuning a microscope system of claim 6, wherein the first adjustment structure comprises a fixed assembly;
the adjusting method comprises the following steps: and when the second image is coincident with the first image, fixing the carrying platform by using the fixing component.
11. The method of claim 5, wherein the second adjusting structure comprises an adjusting plate and a plurality of second adjusting members, the supporting module comprises a base disposed on the adjusting plate, the plurality of second adjusting members are spaced apart from each other and movably connected to the base and the adjusting plate, and the second adjusting members are adjusted to drive the adjusting plate so that the surface of the reactor and the object-carrying surface satisfy the predetermined positional relationship.
12. The method of claim 11, wherein adjusting the second adjustment structure to bring the surface of the reactor and the carrier surface into a predetermined positional relationship comprises:
adjusting the second adjusting member so that an absolute value of a difference between the first distance and the second distance is smaller than a first set value and an absolute value of a difference between the third distance and the fourth distance is smaller than a second set value,
the first distance is a distance from the camera to a first location on the reactor surface,
the second distance is a distance from the camera to a second location on the reactor surface,
the third distance is a distance from the camera to a third location on the reactor surface,
the fourth distance is a distance from the camera to a fourth location on the reactor surface,
a line connecting the first position and the second position is non-parallel to a line connecting the third position and the fourth position.
13. The method of tuning a microscope system according to claim 11, the method comprising:
adjusting part of the second adjusting member so that a distance between a first position and a second position on the surface of the reactor in the direction of reflected light is smaller than a first set value;
and fixing a part of the second regulating member, and regulating another part of the second regulating member so that a distance between a third position and a fourth position on the reactor surface in the direction of reflected light is smaller than a second set value.
14. A microscope system calibrated by any of the methods of claims 1-13.
15. A sequencing system comprising the microscope system of claim 14.
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