CN105911684B - A kind of positioning device and localization method for carrying out target positioning between different microscopes - Google Patents
A kind of positioning device and localization method for carrying out target positioning between different microscopes Download PDFInfo
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- CN105911684B CN105911684B CN201510524424.2A CN201510524424A CN105911684B CN 105911684 B CN105911684 B CN 105911684B CN 201510524424 A CN201510524424 A CN 201510524424A CN 105911684 B CN105911684 B CN 105911684B
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Abstract
The present invention provides a kind of positioning devices and localization method for carrying out target positioning between different microscopes.Apparatus of the present invention include positioning glass slide and polychrome colour bar scale plate, the wherein described multicolour scale plate is fixed in a main surface of the positioning glass slide, the polychrome colour bar scale plate is equipped with one or more scale cog regions, and the scale cog region includes the scale grid being staggeredly made of mass-tone graduation mark L1 and polychrome graduation mark L2.Using apparatus of the present invention, target micro-dissections region can be converted into specific coordinate readings, to be quickly and accurately positioned target cutting region in laser microdissection system, and then target is rapidly and accurately found, then can accurately carry out the subsequent operation such as being cut by laser.
Description
Technical field
The present invention relates to biotechnology, more particularly to a kind of positioning for carrying out target positioning between different microscopes
Device and localization method.
Background technology
Laser microprobe dating technology is using laser to biological sample (tissue, cell cluster, the unicellular, dye under microscope
Colour solid, chromosome segment etc.) carry out the technologies of contactless micro-dissections and separation.The technology is real using the ejection of inverse gravity optical pressure
Now to the active collection of sample, whole process can be accurately positioned without any Mechanical Contact, no thermal damage, system, therefore can be true
Protect obtain sample is pollution-free, high-purity.Microdissection technology is one of the important channel for realizing accurate molecular biological analysis,
It is also one of the hot technology that progress DNA, RNA, protein equimolecular biological study are the most commonly used at present.
Laser microdissection system includes microscopic system, under the microscope can it is convenient, fast, accurately from cell
The complete single cell of form or cell mass are obtained in natural tissues environment, protects it from the pollution of surrounding tissue, and can be compared with
Retain surrounding tissue well, to further investigate.Microdissection technology plays important in the comparative studies of specific cells subgroup
Effect, it is also presently the most the technology for successfully solving cell heterogeneity in tissue.The technology is multiple at present
Succeed application in research field, such as animal-plant gene expression analysis, the comparative studies of protein group, oncology studies, life
Grow with embryonic development research, metabolite analysis etc..
It is a variety of aobvious in the market with the fast development of molecular biology research and the extensive use of laser microprobe dating technology
Micro- diced system comes into being.But as high-grade, precision instrument, the market price of the micro-dissection system of complete set is still
In more high-level, it might even be possible to say that it belongs to expensive instrument and equipment, be difficult to be equipped with by common lab.Therefore, a set of
Micro-dissection system is usually multiple laboratories or research institution shares or even the sample of different regions converges to micro-dissections
It is detected at where system, this just proposes researcher the requirement for carrying out instrument reservation in advance, when increasing scientific research
Between cost.
In addition, the limitation due to being equipped with equipment, in current micro-dissection system the configuration of microscopic system be also difficult to full
The Research Requirements of sufficient people, and many micro-dissection systems do not have coordinate scale, this gives the scientific research that need to carry out micro-dissections operation
People finder's target cutting region brings prodigious difficulty.And if by customizing the special microscope system for reaching Research Requirements
The method for uniting and being equipped with certain coordinate scale solves the difficulty, then can additionally increase very big outfit cost of equipment.
It, can be simple, quick, low there is an urgent need in the art to develop in view of the disadvantages described above of existing laser microprobe dating technology
The positioning device for carrying out target positioning between different microscopes of cost.
Invention content
The object of the present invention is to provide a kind of simple, quickly, inexpensive progress target positioning between being used for different microscopes
Positioning device and localization method.
The first aspect of the present invention provides a kind of positioning device for carrying out target positioning between different microscopes, packet
It includes：One positioning glass slide and a polychrome colour bar scale plate, wherein polychrome colour bar scale plate are fixed on a master of positioning glass slide
On surface, and the 10%-90% that the area of polychrome colour bar scale plate is the main surface area for positioning glass slide；
Polychrome colour bar scale plate is equipped with one or more scale cog regions, scale cog region include by mass-tone graduation mark L1 and
The scale grid that polychrome graduation mark L2 is staggeredly constituted, and press " L1-L2 " arrangement mode interval arranged adjacent；
The color of polychrome graduation mark L2 is different from the color of mass-tone graduation mark L1, and the color category of polychrome graduation mark L2 is
3-10 kinds, and the distance between two mass-tone graduation mark L1 of arbitrary neighborhood are 0.01cm-0.5cm.
In another preferred example, the device is additionally provided with the scale for being illustrated to scale reading and measures explanation
Area, wherein the scale measurement illustrates that area is set on the positioning glass slide or is set on the polychrome colour bar scale plate.
In another preferred example, the area of the polychrome colour bar scale plate is the main surface area of the positioning glass slide
20%-80%.
In another preferred example, the polychrome graduation mark L2 is one or more by 3-10 in laterally and/or longitudinally upper composition
The polychrome graduation mark cycle group that polychrome graduation mark L2 described in item is constituted, and in each polychrome graduation mark cycle group, institute
Polychrome graduation mark L2 is stated to be arranged by identical color sequence.
In another preferred example, the color category of the polychrome graduation mark L2 is 3,4,5 or 6 kind.
In another preferred example, in a scale cog region, two polychrome graduation mark L2 of arbitrary neighborhood
The distance between the distance between two mass-tone graduation mark L1 of arbitrary neighborhood it is equal.
In another preferred example, the line width of the mass-tone graduation mark L1 and polychrome graduation mark L2 are two of arbitrary neighborhood
The 1/50 to 1/4 of the distance between the mass-tone graduation mark L1.
In another preferred example, the scale cog region is rectangle, round or ellipse.
The second aspect of the present invention, provides a kind of microscopic slide kit, and the kit includes n for showing
Any positioning device in the specimen slide and first aspect present invention of micro mirror observation, and the specimen slide
Specification and it is described positioning glass slide specification be it is identical, wherein n be 5-1000 positive integer.
The third aspect of the present invention provides a kind of localization method for carrying out target positioning between different microscopes, packet
Include step：
(a) specimen slide is positioned under the first microscope and is observed, be wherein placed with sample in specimen slide,
Sample contains the target of pending laser microprobe dating；
(b) target of pending laser microprobe dating is found under the first microscope, and records target in the first microscope
Under microscope coordinate readings, be denoted as the first coordinate readings；
(c) specimen slide is removed from the first microscope, and is changed to described for carrying out mesh between different microscopes
The positioning device of position is demarcated, the specification of positioning glass slide and the specification of specimen slide wherein in positioning device are consistent；
(d) it is based on positioning device, determines respective coordinates of first coordinate readings in the scale cog region of positioning device, is remembered
For the second coordinate readings；
(e) positioning device is removed from the first microscope, and positioning device is positioned over for carrying out laser microprobe dating
The second microscope under；
(f) corresponding position of second coordinate readings in scale cog region is found under the second microscope, determines that target exists
Corresponding field of view in second microscope；
(g) positioning device is removed from the second microscope, and is changed to the specimen slide described in step (a) so that
Target is located at the second microscopical field of view, to realize the positioning to target.
It should be understood that within the scope of the present invention, above-mentioned each technical characteristic of the invention and have in below (eg embodiment)
It can be combined with each other between each technical characteristic of body description, to form a new or preferred technical solution.As space is limited, exist
This no longer tires out one by one states.
Description of the drawings
Fig. 1 is the structural schematic diagram of the positioning device in embodiment 1；
Fig. 2 is the structural schematic diagram of the positioning device in embodiment 2.
Specific implementation mode
The present inventor after extensive and in-depth study, develop for the first time it is a kind of it is simple in structure, maneuverable for
It is fixed can rapidly and accurately to carry out target area using the positioning device for the positioning device that target positioning is carried out between different microscopes
Position is easy to implement the quick lookup for single target tumour cell and carries out subsequent operation (such as micro-dissections) to it, in turn
Convenient for subsequently being analyzed (full exon such as unicellular to tumour or Whole genome analysis) to target, complete on this basis
The present invention.
Term
As used herein, term " scale grid " refers in horizontal and vertical graduation mark (including mass-tone graduation mark L1 and more
Color graduation mark L2) interlock the rectangular mesh constituted.
It should be noted that in the claim and specification of this patent, such as first and second or the like relationship
Term is only used to distinguish one entity or operation from another entity or operation, without necessarily requiring or implying
There are any actual relationship or orders between these entities or operation.Moreover, the terms "include", "comprise" or its
Any other variant is intended to non-exclusive inclusion so that including the processes of a series of elements, method, article or
Equipment includes not only those elements, but also includes other elements that are not explicitly listed, or further include for this process,
Method, article or the intrinsic element of equipment.In the absence of more restrictions, being wanted by what sentence " including one " limited
Element, it is not excluded that there is also other identical elements in the process, method, article or apparatus that includes the element.
Positioning device
The present invention provides a kind of positioning device for carrying out target positioning between different microscopes, which includes：It is fixed
Position glass slide 1, polychrome colour bar scale plate 2.Wherein, polychrome colour bar scale plate 2 is fixed on a main surface of positioning glass slide 1
On.
Preferably, the positioning device further includes that the scale measurement for being illustrated to scale reading illustrates area 4, scale
Measurement illustrates that area 4 is set on positioning glass slide 1 or is set on polychrome colour bar scale plate 2.
In the present invention, (such as 1-10, preferably 1-5 is a, more preferably equipped with one or more for polychrome colour bar scale plate 2
1-3) scale cog region 3, each scale cog region 3 includes the scale being staggeredly made of mass-tone graduation mark L1 and polychrome graduation mark L2
Grid, and press " L1-L2 " arrangement mode interval arranged adjacent.
In the present invention, typically, the color of polychrome graduation mark L2 is different from the color of mass-tone graduation mark L1, and polychrome is carved
The color category for spending line L2 is 3-10 kinds, and the distance between two mass-tone graduation mark L1 of arbitrary neighborhood are 0.01cm-
0.5cm。
In the present invention, two adjacent the distance between mass-tone graduation marks are not particularly limited.In general, can define two
The distance between adjacent mass-tone graduation mark is 1 (length unit) in reference axis, defines colorful graduation mark and adjacent mass-tone
The distance between graduation mark is 0.5 (length unit) in reference axis.
In the present invention, the polychrome graduation mark L2 can be in the laterally and/or longitudinally upper one or more that constitutes by 3-10 items
The polychrome graduation mark cycle group that polychrome graduation mark L2 is constituted, and in each polychrome graduation mark cycle group, polychrome graduation mark L2
It is arranged by identical color sequence and (is such as repeated " blue-red-yellow-green-purple " color sequence).
In the present invention, it is preferred to scale cog region 3 be rectangle, round or ellipse.
In the present invention, it is preferred in a scale cog region 3, two polychrome graduation mark L2 of arbitrary neighborhood
The distance between the distance between two mass-tone graduation mark L1 of arbitrary neighborhood equal, mass-tone graduation mark L1 and polychrome graduation mark
The line width of L2 is the 1/50 to 1/4 of the distance between two mass-tone graduation mark L1 of arbitrary neighborhood.
In another preferred example, the area of polychrome colour bar scale plate 2 is the 20%- for the main surface area for positioning glass slide 1
80%.
In another preferred example, the color category of polychrome graduation mark L2 is not particularly limited, usually >=3 kinds.Representative kind
Class includes (but being not limited to)：3,4,5 or 6 kind.
In another preferred example, the color of polychrome graduation mark L2 is purple, green, yellow, red, blue, and polychrome graduation mark L2 is in cross
To with identical polychrome graduation mark cycle group is constituted on longitudinal direction, wherein polychrome graduation mark L2 is pressed in polychrome graduation mark cycle group
The color sequence of " purple-green-yellow-red-blue " arranges.
In another preferred example, the line width of the mass-tone graduation mark and polychrome graduation mark is described in two of arbitrary neighborhood
The 1/25 to 1/8 of the distance between mass-tone graduation mark.
In another preferred example, the scale cog region 3 is rectangle.
In another preferred example, the polychrome colour bar scale plate 2 is equipped with the scale cog region 3 of 2-3 arranged adjacent.
In another preferred example, in the scale cog region 3 of the arranged adjacent, the distance between mass-tone graduation mark L1 is phase
It is same or different that (the distance between each L1 is respectively (i) 0.02,0.05 and 0.10cm in such as 3 scale cog regions 3；Or
(ii) 0.05,0.10 and 0.15cm).
In the present invention, the material of positioning glass slide 1 is not particularly limited, it is ensured that light transmission can be any use
Glass slide material.Representative material includes (but being not limited to)：Glass, plastics.In general, the material is transparent, and
And it is colourless or has certain color.Certainly, the material that the present invention positions glass slide 1 can also be opaque or translucent
's.
Certainly, in another preferred example, the present invention position glass slide 1 material and specimen slide material be it is identical or
Essentially identical.
In conjunction with the attached drawing positioning device that the present invention is further explained.
A kind of typical positioning device of the present invention is as shown in Figure 1.Positioning device in Fig. 1 includes：Positioning glass slide 1,
Polychrome colour bar scale plate 2 and scale measurement illustrate area 4.Wherein positioning glass slide 1 is cytelligen sheet glass, with cell sample
Glass slide specification used in smear is identical, and scale measurement illustrates that area 4 is set on positioning glass slide 1, and polychrome colour bar scale plate 2 is solid
Due in a main surface of positioning glass slide 1, polychrome colour bar scale plate 2 is equipped with multiple scale cog regions 3, all multiple scales
Cog region 3 constitutes long 1.5cm, the rectangle of wide 1.5cm.Each the color of the mass-tone graduation mark L1 in scale cog region 3 is
The color of black, polychrome graduation mark L2 is purple, green, yellow, red, blue, and polychrome graduation mark L2 is identical in horizontal and vertical upper composition
Polychrome graduation mark cycle group, wherein in polychrome graduation mark cycle group polychrome graduation mark L2 press " purple-green-yellow-red-blue " face
Order of colors arranges, and the distance of two mass-tone graduation marks of arbitrary neighborhood is 0.1cm, two polychrome graduation marks of arbitrary neighborhood
Distance is 0.1cm, and the mass-tone graduation mark of arbitrary neighborhood is 0.05cm at a distance from polychrome graduation mark.
The typical positioning device of another kind of the present invention is as shown in Figure 2.Positioning device in Fig. 2 includes：Position glass slide
1, polychrome colour bar scale plate 2 and scale measurement illustrate area 4.Wherein positioning glass slide 1 is the positioning glass slide 1 in Fig. 1, and scale is surveyed
Amount illustrates that area 4 is set on positioning glass slide 1, and polychrome colour bar scale plate 2 is fixed in a main surface of positioning glass slide 1,
Polychrome colour bar scale plate 2 is equipped with multiple scale cog regions 3, and all multiple scale cog regions 3 constitute long 4.5cm, the length of wide 2.0cm
It is rectangular.The color of mass-tone graduation mark L1 in each scale cog region 3 is black, the color of polychrome graduation mark L2 be it is purple, green,
It is yellow, red, blue, and polychrome graduation mark L2 upper constitutes identical polychrome graduation mark cycle group horizontal and vertical, wherein polychrome scale
Polychrome graduation mark L2 is arranged by the color sequence of " purple-green-yellow-red-blue " in line cycle group, two mass-tone scales of arbitrary neighborhood
The distance of line is 0.1cm, and the distance of two polychrome graduation marks of arbitrary neighborhood is 0.1cm, the mass-tone scale of arbitrary neighborhood
Line is 0.05cm at a distance from polychrome graduation mark.
Kit
The present invention also provides a kind of microscopic slide kits, including the n samples for micro- sem observation to carry glass
Piece and the positioning device, and the specification of specimen slide and the specification of the glass slide in the positioning device are phases
With, wherein n is the positive integer of 5-1000, and preferably n is 10-200.
Localization method and subsequent operation
Target positioning is carried out the present invention also provides a kind of positioning device using the present invention and carries out subsequent operation (such as
Laser cutting) method, it includes step：
(a) specimen slide is positioned under the first microscope and is observed, wherein being placed in the specimen slide
Sample, the sample contain the target of pending laser microprobe dating；
(b) target of the pending laser microprobe dating is found under first microscope, and records the target
Microscope coordinate readings under the first microscope, are denoted as the first coordinate readings；
(c) specimen slide is removed from first microscope, and be changed to described in different microscopes
Between carry out the positioning device of target positioning, wherein the specification of the positioning glass slide 1 in the positioning device and the sample
The specification of glass slide is consistent；
(d) it is based on the positioning device, determines that first coordinate readings are identified in the scale of the positioning device
Respective coordinates in area 3 are denoted as the second coordinate readings；
(e) positioning device is removed from first microscope, and the positioning device is positioned over for carrying out
Under second microscope of laser microprobe dating；
(f) correspondence position of second coordinate readings in the scale cog region 3 is found under second microscope
It sets, determines the target corresponding field of view in second microscope；
(g) positioning device is removed from second microscope, and is changed to the sample described in step (a) and carries glass
Piece so that the target is located at the described second microscopical field of view, to realize the positioning to the target.
In general, second microscope is the equipment for carrying out the operation such as being cut by laser.By the positioning of the present invention
Device can quickly and accurately position the targeted target (such as cancer cell) of the operation such as pending laser cutting, and help
In significant decrease error probability, therefore the working efficiency of operating personnel can be greatly improved and reduce working strength.
In another preferred example, the method for the present invention further includes：In step (f), second coordinate readings are recorded
Microscope coordinate readings under two microscopes, are denoted as third coordinate readings.
In another preferred example, method of the invention further includes：In step (h), to the pending laser capture microdissection
The target of cutting carries out laser microprobe dating.
In the present invention, it is suitble to the sample for being observed and being carried out micro-dissections with the method for the present invention to be not particularly limited,
Representative sample includes (but being not limited to)：Tissue samples, cell sample.
In another preferred example, preferred target includes tumour cell, normal cell.
In the present invention, the sample can come from a variety of different sources, and representative source includes (but and unlimited
In)：Mammal (such as people), bacterium, fungi, actinomyces, plant.
The method of the present invention can be diagnostic or nondiagnostic.In a kind of preference, method right and wrong of the invention
It is diagnostic and non-therapeutic.
Main advantages of the present invention include：
(a) it carries out target using the positioning device of the present invention and positions the quick lookup that may be implemented to target, than generally looking into
It looks for and has saved at least about time of 70%-90%, improve work efficiency.
(b) it is equipped with the scale measurement for being illustrated to scale reading and illustrates that area 4, person easy to operation quickly read target
Corresponding coordinate.
(c) enable the color of adjacent graduation mark on polychrome colour bar scale plate 2 different, convenient for distinguishing graduation mark.
(d) it is 1 to define the distance between two adjacent mass-tone graduation marks in reference axis, defines colorful graduation mark and phase
The distance between adjacent mass-tone graduation mark is 0.5 in reference axis, convenient for giving target label coordinate.
(e) positioning device and method of the invention can be additionally used in the differentiated different type microscope or of the same race of coordinate position
The quick positioning of target area between type different brands microscope.
Present invention will be further explained below with reference to specific examples.It should be understood that these embodiments are merely to illustrate the present invention
Rather than it limits the scope of the invention.In the following examples, the experimental methods for specific conditions are not specified, usually according to conventional strip
Part, or according to the normal condition proposed by manufacturer.Unless otherwise stated, otherwise percentage and number are weight percent and weight
Number.
Embodiment 1
The present embodiment positions target using positioning device shown in FIG. 1.In the present embodiment, target cell DAPI
Positive, CD45 feminine genders, the CEP-8 positives tumour cell.Target cell is found in the case where Nikon 80i is just setting 40 power microscope of fluorescence,
The cell is confirmed for the cell is moved to field of microscope centre after target cell, is recorded the abscissa of microscope at this time and is indulged
Coordinate readings are (x:52.8 y:103.6) above-mentioned positioning device, is changed, area 4 is illustrated by scale measurement, reads and records this
When target cell scale cog region 3 in polychrome colour bar scale plate 2 corresponding coordinate, the self-defined coordinate of target cell can give birth to
At.
In the case where the Zeiss of micro-dissection system is inverted 40 power microscope of fluorescence, above-mentioned positioning device is put into objective table, is looked for
To after the self-defined coordinate of upper step, it is recorded in the microscope coordinate position of target cell under the microscope.
Above-mentioned positioning device is gently removed, sample slide is put, return to previous step in micro-dissection system record it is aobvious
Micro mirror coordinate position, has been quickly found out target cell.The target cell can subsequently be further looked at or be obtained by micro-dissections
Take the unicellular full exon of unicellular progress or Whole genome analysis etc..
Wherein, it is just being set from Nikon 80i and is finding target cell under 40 power microscope of fluorescence and count, until in micro-dissection system
The target cell to be found under Zeiss inversion 40 power microscope of fluorescence to stop, the average used time of 6 bit manipulation personnel is 3 minute/cells,
Compared with comparative example 1, the used time saves 98.57%.Target cell searches success rate 100%, and compared with comparative example 1, success rate is high
75%.
Comparative example 1
Embodiment 1 is repeated, the difference lies in that omitting using positioning device used in embodiment 1.
As a result：It is just being set from Nikon 80i and is finding target cell under 40 power microscope of fluorescence and count, until in micro-dissection system
The target cell is found under Zeiss inversion 40 power microscope of fluorescence to stop, the average used time of 6 bit manipulation personnel is that 3.5 hours/are thin
Born of the same parents, wherein 4 bit manipulation people finder's fall short cells, target cell searches success rate 25%.
Embodiment 2
The present embodiment positions target using positioning device shown in Fig. 2.In the present embodiment, certain colleges and universities of the areas A grind
Study carefully the cell there are one specific type in institute's new discovery sample, it need to this be new with research institution's communication and discussion of the areas B same type
It was found that cell.
Certain research institute of colleges and universities of the areas A is coming 40 times of microscopic observations of card microscope to target cell, records target cell and exists
Microscope coordinate (x when field of microscope centre:54, y:105).Above-mentioned positioning device is changed, area is illustrated by scale measurement
4, the self-defined coordinate for recording target cell scale cog region 3 in polychrome colour bar scale plate 2 at this time is (x:10, y:4).It will
Sample slide and the self-defined coordinate (x of above-mentioned positioning device and target cell in above-mentioned positioning device:10, y:4) information one
It is same to post to B area studies mechanism.
B area studies mechanism is according to the self-defined coordinate (x of target cell:10, y:4) information, the Buddhist nun used in its laboratory
Self-defined coordinate (x of the target cell in above-mentioned positioning device is found on health microscope:10, y:4), record at this time sit by microscope
Mark reading (x:54.5 y:107.5).Sample slide is changed, microscope coordinate position is returned to, has been quickly found out target cell.
Wherein, B area studies mechanism is from taking sample slide and target cell information begins look for target cell and counts, until
It finds the target cell under Nikon microscope to stop, the average used time of 6 bit manipulation personnel is 30 seconds.Compared with comparative example 2, the used time
Save 99.59%.
Comparative example 2
Embodiment 2 is repeated, the difference lies in that omitting using positioning device used in embodiment 2.
As a result：B area studies mechanism from taking sample slide and target cell information begins look for target cell and counts, until
It finds the target cell under Nikon microscope to stop, the average used time of 6 bit manipulation personnel is 2 hours.
All references mentioned in the present invention is incorporated herein by reference, independent just as each document
It is incorporated as with reference to such.In addition, it should also be understood that, after reading the above teachings of the present invention, those skilled in the art can
To be made various changes or modifications to the present invention, such equivalent forms equally fall within model defined by the application the appended claims
It encloses.
Claims (8)
1. a kind of method carrying out target positioning between different microscopes, which is characterized in that including step：
(a) specimen slide is positioned under the first microscope and is observed, wherein it is placed with sample in the specimen slide,
The sample contains the target of pending laser microprobe dating；
(b) target of the pending laser microprobe dating is found under first microscope, and records the target
Microscope coordinate readings under one microscope, are denoted as the first coordinate readings；
(c) specimen slide is removed from first microscope, and be changed to for carrying out mesh between different microscopes
The positioning device of position is demarcated, wherein the specification of the specification and the specimen slide of the positioning glass slide in the positioning device is
Consistent；
(d) it is based on the positioning device, determines pair of first coordinate readings in the scale cog region of the positioning device
Coordinate is answered, the second coordinate readings are denoted as；
(e) positioning device is removed from first microscope, and the positioning device is positioned over for carrying out laser
Under second microscope of micro-dissections；
(f) corresponding position of second coordinate readings in the scale cog region is found under second microscope, really
The fixed target corresponding field of view in second microscope；
(g) positioning device is removed from second microscope, and is changed to the specimen slide described in step (a),
So that the target is located at the described second microscopical field of view, to realize the positioning to the target；
In the step (c) between different microscopes carry out target positioning positioning device include：One positioning glass slide and
One polychrome colour bar scale plate；
The polychrome colour bar scale plate is fixed in a main surface of the positioning glass slide, and the polychrome colour bar scale plate
Area be it is described positioning glass slide main surface area 10%-90%；
The polychrome colour bar scale plate is equipped with one or more scale cog regions, and the scale cog region includes by mass-tone graduation mark
The scale grid that L1 and polychrome graduation mark L2 is staggeredly constituted, wherein the face of the mass-tone graduation mark L1 and the polychrome graduation mark L2
Color is different, and presses " L1-L2 " arrangement mode interval arranged adjacent；
The color of the polychrome graduation mark L2 is different from the color of mass-tone graduation mark L1, and the face of the polychrome graduation mark L2
Color type is 3-10 kinds；
Also, the distance between two described mass-tone graduation mark L1 of arbitrary neighborhood are 0.01cm-0.5cm.
2. according to the method described in claim 1, it is characterized in that, the device is additionally provided with for being said to scale reading
Bright scale measurement illustrates area, wherein the scale measurement illustrates that area is set on the positioning glass slide or is set to described more
On color bar scale plate.
3. according to the method described in claim 1, it is characterized in that, the area of the polychrome colour bar scale plate carries for the positioning
The 20%-80% of the main surface area of slide.
4. according to the method described in claim 1, it is characterized in that, the polychrome graduation mark L2 is in laterally and/or longitudinally upper structure
At the polychrome graduation mark cycle group that one or more polychrome graduation mark L2 described in 3-10 items is constituted, and in each polychrome
In graduation mark cycle group, the polychrome graduation mark L2 is arranged by identical color sequence.
5. method according to any one of claims 1-4, which is characterized in that the color category of the polychrome graduation mark L2 is
3,4,5 or 6 kind.
6. method according to any one of claims 1-4, which is characterized in that in a scale cog region, arbitrarily
Between the distance between two adjacent described polychrome graduation mark L2 and two mass-tone graduation mark L1 of arbitrary neighborhood away from
From equal.
7. method according to any one of claims 1-4, which is characterized in that the mass-tone graduation mark L1 and polychrome graduation mark
The line width of L2 is the 1/50 to 1/4 of the distance between two described mass-tone graduation mark L1 of arbitrary neighborhood.
8. method according to any one of claims 1-4, which is characterized in that the scale cog region be rectangle, circle or
Ellipse.
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| CN106442324A (en) * | 2016-11-01 | 2017-02-22 | 昆明泰来光学科技开发有限公司 | Multi-waveband LED reflected/transmitted illumination objective table |
| CN110560930B (en) * | 2019-09-16 | 2021-11-16 | 深圳泰软软件科技有限公司 | Cutting method and device for blood spot of blood sampling card and readable storage medium |
| CN112798475A (en) * | 2020-12-22 | 2021-05-14 | 西安科技大学 | Method, system and device for monitoring diffusion area of grouting slurry in rock-soil mass |
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| US20020192650A1 (en) * | 2001-05-30 | 2002-12-19 | Amorese Douglas A. | Composite arrays |
| CN2729704Y (en) * | 2004-09-07 | 2005-09-28 | 中国人民解放军第三军医大学 | Organisms macromolecule microarray positioner |
| CN201035215Y (en) * | 2007-01-19 | 2008-03-12 | 马青松 | Glass carrier special for pathological mechanism |
| CN103627619A (en) * | 2012-08-23 | 2014-03-12 | 徐元诚 | Grid positioning sheet capable of detecting sperms quickly |
| CN202794697U (en) * | 2012-12-05 | 2013-03-13 | 崔文钰 | Slide glass sheet with scale |
| CN203054338U (en) * | 2013-01-08 | 2013-07-10 | 复旦大学 | Slide glass with grid reference lines |
| CN203037919U (en) * | 2013-01-25 | 2013-07-03 | 浙江师范大学 | Novel cover slip with mark |
| CN205027969U (en) * | 2015-08-24 | 2016-02-10 | 上海宝藤生物医药科技股份有限公司 | A positioner that is used for carrying on between different microscopes object positioning |
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