CN107390356A - The method focused on automatically based on representation of laser facula - Google Patents
The method focused on automatically based on representation of laser facula Download PDFInfo
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- CN107390356A CN107390356A CN201710749519.3A CN201710749519A CN107390356A CN 107390356 A CN107390356 A CN 107390356A CN 201710749519 A CN201710749519 A CN 201710749519A CN 107390356 A CN107390356 A CN 107390356A
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- laser facula
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- G—PHYSICS
- G02—OPTICS
- G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
- G02B21/00—Microscopes
- G02B21/24—Base structure
- G02B21/241—Devices for focusing
- G02B21/244—Devices for focusing using image analysis techniques
-
- G—PHYSICS
- G02—OPTICS
- G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
- G02B21/00—Microscopes
- G02B21/24—Base structure
- G02B21/241—Devices for focusing
-
- 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
- G02B21/361—Optical details, e.g. image relay to the camera or image sensor
-
- 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
- G02B21/365—Control or image processing arrangements for digital or video microscopes
Abstract
The invention discloses a kind of method focused on automatically based on representation of laser facula.This method includes：(1) light beam is expanded；(2) beam and focus is corrected as semicircle, and overlaps its center of circle and the center of circle of object lens light hole；(3) focal curve figure is established, the peak value in figure is in X-axis midpoint；(4) using method forming curves figure described in step (3), by curve map peak value and X-axis center position relation, judgement sample defocus distance and direction；(5) processing terminal driving focus adjusting mechanism motion, focuses on sample.The inventive method algorithm is simple, and focusing speed is much better than the ordinary gamma type of focusing, and can draw defocus direction and distance according to piece image.
Description
Technical field
The invention belongs to the automatic focus area of light microscope, and in particular to one kind is focused on automatically based on representation of laser facula
Method.
Background technology
In microscope imaging, due to the shorter factor of object lens depth of focus, after focusing on one point, the mistake of other points is moved to
Because the horizontality of objective table or sample is bad etc., reason, image can thicken in journey.
To solve this problem, it is common based on types of focusing such as gradation of images because it needs to gather multiple image, calculate
Method is complicated and direction and the distance of defocus can not be being provided according to piece image can.
Therefore the mode of efficiency factor gradation of image focusing is considered in some requirement high speeds in real time system and is not applied to.
The content of the invention
For above-mentioned deficiency of the prior art, the present invention provides a kind of side focused on automatically based on representation of laser facula
Method, can effectively solve existing MIcrosope image focus method complexity, and defocus direction and distance can not be drawn according to piece image
The problem of.
A kind of method focused on automatically based on representation of laser facula, is comprised the following steps：
(1) laser beam is expanded using laser beam expander, and the angle of divergence for expanding rear light beam is less than 1.2mard；
(2) beam and focus after expanding is modified to semicircle, and makes its normal incidence into microscopical object lens, adjust light
Road, overlap the hot spot center of circle and the center of circle of object lens light hole；
(3) it is provided with the digital imaging system being connected with processing terminal in microscope, processing terminal is by digital imagery system
The sample image positioned at focal point collected of uniting is converted into two-dimensional matrix, then two-dimensional matrix is converted into one-dimensional matrix, filters,
Then using one-dimensional matrix element position as X-axis, corresponding element value is Y-axis, forms focal curve figure, adjustment digital imaging system position
Put, the peak value in focal curve figure is in X-axis midpoint；
(4) digital imaging system continues to gather sample image, by method forming curves figure described in step (3), passes through
Curve map peak value and X-axis center position relation, judgement sample defocus distance and direction；
(5) focus adjusting mechanism for being equipped with and being connected with processing terminal is set up separately in microscope Z axis trimming part, processing terminal is according to step
(4) the judged result driving focus adjusting mechanism motion in, focuses on sample, then carries out the detection of next sample again.
Further, beam and focus is modified by the way of blocking in step (2), so that it becomes semicircle.
Further, digital imaging system is CMOS camera or CCD camera in step (3).
Further, it is provided with band pass filter before digital imaging system.
Further, defocus distance and the deterministic process in direction are in step (4)：
If sample, above focus, curve map peak can be on the right side of X-axis central point, and the bigger peak value of defocus is from X-axis
Central point is more remote, conversely, when sample is below focus, peak can be on the left of X-axis central point, and defocus is bigger, and peak value is from X
Axis center point is more remote.
Further, digital imaging system and focus adjusting mechanism are connected by way of communication connection with processing terminal.
Further, focus adjusting mechanism is Z axis piezoelectric position moving stage, Z axis stepper motor displacement platform, drives thin quasi- Jiao on microscope
The stepper motor of helical rotation or the piezoelectricity object lens being arranged on micro objective；And the stepper motor and thin accurate burnt spiral
Between be provided with cylindrical adaptor.
Beneficial effects of the present invention are：
When carrying out image focusing using the inventive method, focal curve figure is first established, when sample, which is in, to be focused on, is adjusted to
As system position, peak value in curve map is set to be located at X-axis central spot, it is determined that sample peak value present position when focusing on, and made with this
On the basis of, carry out subsequent sample detection；Its principle is：After semicircular laser light incident object lens, if sample is in Jiao of object lens
Point, the hot spot that digital imaging system is collected are a circular light spot, the digital imaging system when sample is above object focal point
The light spot image collected is right semi-circle and the more remote graphics area of focal point is bigger, conversely, number when sample is below object focal point
The spot pattern that word imaging system collects is left semicircle and its area becomes big with defocus degree and becomes big.
If sample above focus, subsequent sample peak can on the right side of X-axis central point, the bigger peak value of defocus from
X-axis central point is more remote, conversely, when sample is below focus, peak can be on the left of X-axis central point, and defocus is bigger, peak value
It is more remote from X-axis central point, according to the distance and position relationship of subsequent sample detection curve figure peak value and X-axis central point, can pass through
Piece image can accurately draw defocus distance and defocus direction, solve the side that focusing algorithm exports in the case where defocus is justified
To distance and reality defocus direction and range difference away from it is larger the problem of.
When carrying out microscope example observation using the inventive method, 0.05s can be most less than soon by completing a focusing, can
Ensure that the image that digital imaging system detects during sample moves is in focus state all the time, ensure that image is located always
In clear state, the detection efficiency of sample is greatly improved.
The present invention provides a kind of fast automatic focusing method, and this method need not carry out big change to light path, and have
There is the characteristics of speed is fast, and algorithm computational complexity is low, overcome that conventional images gray scale type of focusing time loss is big and algorithm is answered
The problem of miscellaneous.
Brief description of the drawings
Fig. 1 is autofocus system structural representation；
The auto-focusing schematic diagram of image auto-focusing methods of the Fig. 2 based on laser facula；
Fig. 3 is the algorithm schematic diagram of the different focal positions of the image auto-focusing method based on laser facula；
Fig. 4 is that the image auto-focusing method based on laser facula is used for a wide range of Raman scanning imagery index path；
Fig. 5 is that the image auto-focusing method based on laser facula is used for microscope example observation automatic focus light path figure；
Fig. 6 is the flow chart of the method for the present invention；
Fig. 7 is stepper motor and thin accurate burnt spiral connection diagram.
Wherein, 1, laser；2nd, laser beam expander；3rd, digital imaging system；4th, focus adjusting mechanism；5th, processing terminal；6th, walk
Stepper motor；7th, adaptor；8th, thin accurate burnt spiral.
Embodiment
The embodiment of the present invention is described below, in order to which those skilled in the art understand this hair
It is bright, it should be apparent that the invention is not restricted to the scope of embodiment, for those skilled in the art,
As long as various change in the spirit and scope of the present invention that appended claim limits and determines, these changes are aobvious and easy
See, all are using the innovation and creation of present inventive concept in the row of protection.
Embodiment 1
As shown in figure 1, the autofocus system, including laser 1, laser beam expander 2, microscope and processing terminal 5, and
The focus adjusting mechanism 4 that communication connection mode is connected with processing terminal is provided with microscope；
Wherein, as shown in fig. 7, focus adjusting mechanism can be disposed on the stepper motor 6 at thin accurate burnt spiral 8 on microscope,
Adaptor 7 is provided between stepper motor 6 and thin accurate burnt spiral 8, adaptor 7 is cylindrical shape, and both ends are nested in stepping electricity respectively
In the rotating shaft of machine 6 and thin accurate burnt spiral 8 so that stepper motor 6 can drive thin accurate burnt spiral 8 to rotate by adaptor 7, or may be used also
Think that Z axis piezoelectric position moving stage, Z axis stepper motor displacement platform or the piezoelectricity object lens being arranged on micro objective, processing terminal 5 are
Computer or embedded processing equipment.
As shown in figure 1, be additionally provided with microscope lens barrel by communication connection mode be connected with processing terminal numeral into
As system 3, and filter plate is provided with before digital imaging system 3, digital imaging system 4 is received returning for laser beam
Light echo, digit preference imaging system 3 are CMOS camera or CCD camera, and filter plate is band pass filter.
The method for using above-mentioned autofocus system focused on automatically based on representation of laser facula, is comprised the following steps：
(1) it is directional light to expand light beam by laser beam expander, and the angle of divergence for expanding rear light beam is less than 1.2mard；
(2) using the means blocked, the beam and focus after expanding is modified to semicircle, and it is just being entered by speculum
It is mapped in microscopical object lens, adjusts light path, overlap the hot spot center of circle and the center of circle of object lens light hole；
(3) sample image positioned at focal point that processing terminal collects digital imaging system converts two-dimensional matrix, will
All row summations, make it be converted into one-dimensional matrix in two-dimensional matrix, and it is filtered, and radio-frequency component are removed, then with one
It is X-axis to tie up matrix element position, and corresponding element value is Y-axis, forms focal curve figure, adjustment digital imaging system position, makes Jiao
Peak value in point curve figure is in X-axis midpoint；
(4) digital imaging system continues to gather sample image, by method forming curves figure described in step (3), if
Sample is above focus, then peak can be on the right side of X-axis central point, and the bigger peak value of defocus is more remote from X-axis central point, conversely,
When sample is below focus, peak can be on the left of X-axis central point, and defocus is bigger, peak value (ginseng more remote from X-axis central point
Fig. 3 is examined, A is focus top curve figure in Fig. 3, and B is focal curve figure, and C is focus lower graph)；
(5) judged result Driving Stepping Motor motion of the processing terminal in step (4), focuses on sample, Ran Houzai
Carry out the detection of next sample (with reference to figure 5, Fig. 6).
The index path of the above method is shown in Fig. 5, and its principle is as follows：
As shown in Fig. 2 after semicircular laser light incident object lens, if sample is in the focus of object lens, digital imaging system
The hot spot collected is a circular light spot, the light spot image that digital imaging system collects when sample is above object focal point
For right semi-circle and the more remote graphics area of focal point it is bigger, conversely, digital imaging system collects when sample is below object focal point
Spot pattern become big with defocus degree for left semicircle and its area and become big.
Embodiment 2
The a wide range of automatic focusing of Raman two-dimensional scan imaging is carried out using the methods described of embodiment 1, its index path is shown in Fig. 4,
Detailed process is as follows：
(1) laser beam expanding for being sent Raman excitation radiant using beam expanding lens, and ensure to expand the angle of divergence of rear laser
In below 1.2mrad；
(2) the laser circular light spot after being expanded by the means of blocking is modified to semicircle hot spot, adjusts light path, makes semicircle
In object lens of the shape hot spot by speculum normal incidence to Raman system, wherein the center of circle of semicircle hot spot and the circle of object lens light hole
The heart overlaps；
(3) microscopical Z axis trimming part sets up separately and is equipped with focus adjusting mechanism in Raman system, then will be micro- in Raman system
The manual stage of mirror is replaced by two-dimensional electricity driving displacement platform, and installs digital imaging system additional in the microscope part of Raman system,
Focus adjusting mechanism, two-dimentional electric moveable platform and digital imaging system are connected by way of communication connection with processing terminal；
(5) the automatic focusing principle according to embodiment 1, after algorithm process, processing terminal is according to peak
Defocus distance is provided, then drives focus adjusting mechanism to be adjusted to focus on by sample；
(6) when large area scanning starts, whether computer can be according to the figure judgement sample that imaging system collects poly-
Jiao, if defocus, sample is adjusted to focus state by computer control stepper motor, this point of Raman spectrometer collection after sample focusing
Data simultaneously preserve, and then computer control two-dimensional stage is moved to subsequent point, judges to focus on, if out-focus control stepper motor is adjusted
To focusing, then Raman spectrometer gathered data preservation (is focused on and judged with reference to figure 3).And so on until by certain regular by sample
Product are scanned, and last processing data simultaneously shows Raman two-dimensional scan result.
A focusing is completed in practical application can most be less than 0.05s soon, it is ensured that the institute during sample moves
Into picture be in focus state all the time.
Claims (7)
- A kind of 1. method focused on automatically based on representation of laser facula, it is characterised in that comprise the following steps：(1) laser beam is expanded using laser beam expander, and the angle of divergence for expanding rear light beam is less than 1.2mard；(2) beam and focus after expanding is modified to semicircle, and makes its normal incidence into microscopical object lens, adjust light path, Overlap the hot spot center of circle and the center of circle of object lens light hole；(3) digital imaging system being connected with processing terminal is provided with microscope, processing terminal adopts digital imaging system The sample image positioned at focal point collected is converted into two-dimensional matrix, then two-dimensional matrix is converted into one-dimensional matrix, filtering, then Using one-dimensional matrix element position as X-axis, corresponding element value is Y-axis, forms focal curve figure, adjusts digital imaging system position, The peak value in focal curve figure is set to be in X-axis midpoint；(4) digital imaging system continues to gather sample image, by method forming curves figure described in step (3), passes through curve Figure peak value and X-axis center position relation, judgement sample defocus distance and direction；(5) focus adjusting mechanism for being equipped with and being connected with processing terminal is set up separately in microscope Z axis trimming part, processing terminal is according to step (4) In judged result driving focus adjusting mechanism motion, focus on sample, then carry out the detection of next sample again.
- 2. the image auto-focusing method according to claim 1 based on laser facula, it is characterised in that in step (2) Beam and focus is modified by the way of blocking, so that it becomes semicircle.
- 3. the image auto-focusing method according to claim 1 based on laser facula, it is characterised in that in step (3) The digital imaging system is CMOS camera or CCD camera.
- 4. the image auto-focusing method based on laser facula according to claim 1 or 3, it is characterised in that the number Band pass filter is provided with before word imaging system.
- 5. the image auto-focusing method according to claim 1 based on laser facula, it is characterised in that in step (4) The defocus distance and the deterministic process in direction are：If sample, above focus, curve map peak can be on the right side of X-axis central point, and the bigger peak value of defocus is from X-axis center Point is more remote, conversely, when sample is below focus, peak can be on the left of X-axis central point, and defocus is bigger, and peak value is from X-axis Heart point is more remote.
- 6. the image auto-focusing method according to claim 1 based on laser facula, it is characterised in that it is described numeral into As system and focus adjusting mechanism are connected by way of communication connection with processing terminal.
- 7. the image auto-focusing method according to claim 1 based on laser facula, it is characterised in that the focusing machine Structure is Z axis piezoelectric position moving stage, Z axis stepper motor displacement platform, drives the stepper motor of thin accurate burnt helical rotation on microscope or set The piezoelectricity object lens being placed on micro objective；And it is provided with cylindrical switching between the stepper motor and thin accurate burnt spiral Part.
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Cited By (6)
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| CN108051897A (en) * | 2018-01-17 | 2018-05-18 | 宁波舜宇仪器有限公司 | A kind of micro imaging system and real-time focusing method |
| CN108254853A (en) * | 2018-01-17 | 2018-07-06 | 宁波舜宇仪器有限公司 | A kind of micro imaging system and its real-time focusing method |
| CN108901118A (en) * | 2018-07-04 | 2018-11-27 | 北京大学 | Transparent target body positioning system and its localization method in a kind of lasing ion accelerator |
| CN112399070A (en) * | 2019-08-19 | 2021-02-23 | 生昊 | Method, device, equipment and storage medium for assisting camera in fast focusing |
| CN114114663A (en) * | 2021-02-25 | 2022-03-01 | 广东粤港澳大湾区黄埔材料研究院 | Focal-locking detection system and method of microscope |
| CN117443858A (en) * | 2023-12-26 | 2024-01-26 | 合肥国镜仪器科技有限公司 | Cleaning method and cleaning system for cold field electron source |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN108051897A (en) * | 2018-01-17 | 2018-05-18 | 宁波舜宇仪器有限公司 | A kind of micro imaging system and real-time focusing method |
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| CN112399070A (en) * | 2019-08-19 | 2021-02-23 | 生昊 | Method, device, equipment and storage medium for assisting camera in fast focusing |
| CN112399070B (en) * | 2019-08-19 | 2022-04-19 | 生昊 | Method, device, equipment and storage medium for assisting camera in fast focusing |
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| CN117443858A (en) * | 2023-12-26 | 2024-01-26 | 合肥国镜仪器科技有限公司 | Cleaning method and cleaning system for cold field electron source |
| CN117443858B (en) * | 2023-12-26 | 2024-03-05 | 合肥国镜仪器科技有限公司 | Cleaning method and cleaning system for cold field electron source |
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Application publication date: 20171124 |