CN103217275A - Magnification factor calibration method for microscope - Google Patents
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- CN103217275A CN103217275A CN2013101756948A CN201310175694A CN103217275A CN 103217275 A CN103217275 A CN 103217275A CN 2013101756948 A CN2013101756948 A CN 2013101756948A CN 201310175694 A CN201310175694 A CN 201310175694A CN 103217275 A CN103217275 A CN 103217275A
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Abstract
The invention discloses a magnification factor calibration method for a microscope. The magnification factor calibration method comprises the following steps of: with a standard raster with a known raster pitch as a reference, acquiring a raster image in a microscope to be calibrated; carrying out fast Fourier transform on the acquired image to obtain a spatial frequency spectrum; selecting an interval where certain level of resonant frequency is located from the spatial frequency spectrum, and carrying out local high-resolution discrete Fourier transform to obtain a local high-resolution spatial frequency spectrum; selecting a frequency with a maximum amplitude as a resonant frequency from the local high-resolution spatial frequency spectrum, and dividing the resonant frequency by the level to obtain a base frequency; calculating the size of the standard raster corresponding to the image by using the raster pitch and the base frequency; and calculating the magnification factor of the microscope by using a ratio of the display size of the image to the size of the standard raster corresponding to the image. Compared with the traditional method, the magnification factor calibration method is high in precision, large in calibration range and wide in applicability when being used for calibrating the magnification factor of the microscope.
Description
Technical field
The invention belongs to exact instrument and technical field of measurement and test, be specifically related to a kind of microscope enlargement factor scaling method.
Background technology
Along with the development of microtechnic, microscope has been used for accurately measuring the geometric properties of microstructure, cell, crystal grain and even atom.In said process, obtaining reliably, experimental data requires to demarcate exactly microscopical enlargement factor.At present, be used for the method that the microscope enlargement factor demarcates and generally can be divided into two kinds, grating standardization and microstructure standardization.Wherein the grating standardization is to utilize the image of gathering master grating at microscopically, utilize the corresponding master grating of the display size of image and image size recently demarcate enlargement factor, owing to be difficult to accurately determine the position of grid line, so this method precision is low, limited by grating frequency, calibration range is narrower.Natural particle, virus or enzyme that the microstructure standardization utilizes some to have geometrical property are demarcated microscopical enlargement factor, but these material geometrical properties have certain uncertainty, also are subjected to the influence of the microscope depth of field simultaneously, and the scope of application is little.Therefore, a kind of stated accuracy height, calibration range is big and applicability is wide scaling method are demanded urgently proposing.
Summary of the invention
The present invention one of is intended to solve the problems of the technologies described above at least to a certain extent or provides a kind of useful commerce to select at least.For this reason, the objective of the invention is to propose the microscope enlargement factor scaling method that a kind of calibration range is big, applicability is wide.
For achieving the above object,, comprise the steps: that A. is reference with the master grating of known raster pitch, in the microscope that need are demarcated, gather raster image according to the microscope enlargement factor scaling method of the embodiment of the invention; B. the image of gathering is carried out Fast Fourier Transform (FFT), obtain spatial frequency spectrum; C. in spatial frequency spectrum, select to carry out local high resolving power discrete Fourier transformation and obtain local high resolution space frequency spectrum between the inferior resonance frequency location of certain one-level; D. the frequency of selecting the amplitude maximum in local high resolution space frequency spectrum is as resonance frequency, with resonance frequency divided by the inferior fundamental frequency that obtains of level; E. by the size of the corresponding master grating of raster pitch and fundamental frequency computed image; F. demarcate microscopical enlargement factor by the ratio of the size of the display size of image and the corresponding master grating of image.
In one embodiment of the invention, adopt following formula to carry out local high resolving power discrete Fourier transformation and obtain local high resolution space frequency spectrum:
μ=μ
d,μ
d+D,μ
d+2D,…,μ
u
ν=ν
d,ν
d+D,ν
d+2D,…,ν
u
Wherein, (μ ν) is local high resolution space frequency spectrum to F; M is the pixel count of image x direction, and N is the pixel count of image y direction; (x is that (x y) locates gray-scale value to image coordinate y) to f; D is the frequency increment of local high resolving power Fourier transform, and the size of D requires to adjust according to computational accuracy; μ is an x director space frequency, and ν is a y director space frequency; (μ
d, μ
u) be the frequency separation of the local high resolving power Fourier transform of image x direction, (ν
d, ν
u) be the frequency separation of the local high resolving power Fourier transform of image y direction; I is an imaginary unit; π is a circular constant; E is the natural logarithm truth of a matter.
In one embodiment of the invention, be defined in the frequency of selection amplitude maximum in the local high resolution space frequency spectrum as resonance frequency (μ
m, ν
n), time obtain fundamental frequency (μ with resonance frequency divided by level
0, ν
0), promptly
Wherein, m and n be x and y direction resonance frequency the level time.
In one embodiment of the invention, the definition raster pitch is (P
x, P
y) and fundamental frequency be (μ
0, ν
0), by the size (l of the corresponding master grating of following formula computed image
x, l
y): (l
x, l
y)=(P
xμ
0, P
yν
0).
In one embodiment of the invention, utilize the display size (I of image
x, I
y) and the size (l of the corresponding master grating of image
x, l
y), demarcate microscopical enlargement factor (MAG by following formula
x, MAG
y):
In one embodiment of the invention, among the described step C, select level time high resonance frequency.
In one embodiment of the invention, described grating is orthogonal grating or unidirectional grating.
The present invention utilizes Fourier transform to demarcate microscopical enlargement factor, the precision height; Error when directly determining fundamental frequency adopts and determines senior time resonance frequency, utilizes the method for the relation calculating fundamental frequency of resonance frequency and fundamental frequency again; In addition, determine to dwindle between the inferior resonance frequency location of certain one-level the computation interval of local high resolving power discrete Fourier transformation by the method for utilizing Fast Fourier Transform (FFT) earlier, and then reduce computing time; This method calibration range is big, and applicability is wide.
Additional aspect of the present invention and advantage part in the following description provide, and part will become obviously from the following description, or recognize by practice of the present invention.
Description of drawings
Above-mentioned and/or additional aspect of the present invention and advantage are from obviously and easily understanding becoming the description of embodiment in conjunction with following accompanying drawing, wherein:
Fig. 1 is the process flow diagram of the microscope enlargement factor scaling method of the embodiment of the invention;
Fig. 2 is that pitch is the micro-image of the standard orthogonal grating of 1um;
Fig. 3 is that selected class is between the resonance frequency location of (10,10) on the Fast Fourier Transform (FFT) spectrum.
Embodiment
Describe embodiments of the invention below in detail, the example of described embodiment is shown in the drawings, and wherein identical from start to finish or similar label is represented identical or similar elements or the element with identical or similar functions.Below by the embodiment that is described with reference to the drawings is exemplary, is intended to be used to explain the present invention, and can not be interpreted as limitation of the present invention.
In description of the invention, it will be appreciated that, term " " center "; " vertically "; " laterally "; " length "; " width "; " thickness ", " on ", D score, " preceding ", " back ", " left side ", " right side ", " vertically ", " level ", " top ", " end " " interior ", " outward ", " clockwise ", close the orientation of indications such as " counterclockwise " or position is based on orientation shown in the drawings or position relation, only be that the present invention for convenience of description and simplification are described, rather than the device or the element of indication or hint indication must have specific orientation, therefore orientation structure and operation with specific can not be interpreted as limitation of the present invention.
In addition, term " first ", " second " only are used to describe purpose, and can not be interpreted as indication or hint relative importance or the implicit quantity that indicates indicated technical characterictic.Thus, one or more a plurality of this feature can be expressed or impliedly be comprised to the feature that is limited with " first ", " second ".In description of the invention, the implication of " a plurality of " is two or more, unless clear and definite concrete qualification is arranged in addition.
In the present invention, unless clear and definite regulation and qualification are arranged in addition, broad understanding should be done in terms such as term " installation ", " linking to each other ", " connection ", " fixing ", for example, can be fixedly connected, also can be to removably connect, or connect integratedly; Can be mechanical connection, also can be to be electrically connected; Can be directly to link to each other, also can link to each other indirectly by intermediary, can be the connection of two element internals.For the ordinary skill in the art, can understand above-mentioned term concrete implication in the present invention as the case may be.
In the present invention, unless clear and definite regulation and qualification are arranged in addition, first feature second feature it " on " or D score can comprise that first and second features directly contact, can comprise that also first and second features are not directly contacts but by the contact of the additional features between them.And, first feature second feature " on ", " top " and " above " comprise first feature directly over second feature and oblique upper, or only represent that the first characteristic level height is higher than second feature.First feature second feature " under ", " below " and " below " comprise first feature under second feature and tiltedly, or only represent that the first characteristic level height is less than second feature.
Now in conjunction with the accompanying drawings the specific embodiment of the present invention is described further.
As shown in Figure 1, the microscope enlargement factor scaling method according to the embodiment of the invention may further comprise the steps:
A. the master grating with known raster pitch is reference, gathers raster image in the microscope that need are demarcated.
Particularly, be reference with the master grating of known raster pitch, be placed in the microscope that need are demarcated, it is clear to imaging to adjust focal length, images acquired.
B. the image of gathering is carried out Fast Fourier Transform (FFT), obtain spatial frequency spectrum.
C. in spatial frequency spectrum, select to carry out local high resolving power discrete Fourier transformation and obtain local high resolution space frequency spectrum between the inferior resonance frequency location of certain one-level.
Particularly, in spatial frequency spectrum, select to adopt formula (1) to carry out local high resolving power discrete Fourier transformation and obtain local high resolution space frequency spectrum between the inferior resonance frequency location of certain one-level.
μ=μ
d,μ
d+D,μ
d+2D,…,μ
u
ν=ν
d,ν
d+D,ν
d+2D,…,ν
u
Wherein, (μ ν) is local high resolution space frequency spectrum to F; M is the pixel count of image x direction, and N is the pixel count of image y direction; (x is that (x y) locates gray-scale value to image coordinate y) to f; D is the frequency increment of local high resolving power Fourier transform, and the size of D requires to adjust according to computational accuracy; μ is an x director space frequency, and ν is a y director space frequency; (μ
d, μ
u) be the frequency separation of the local high resolving power Fourier transform of image x direction, (ν
d, ν
u) be the frequency separation of the local high resolving power Fourier transform of image y direction; I is an imaginary unit; π is a circular constant; E is the natural logarithm truth of a matter.
D. the frequency of selecting the amplitude maximum in local high resolution space frequency spectrum is as resonance frequency, with resonance frequency divided by the inferior fundamental frequency that obtains of level.
Particularly, in local high resolution space frequency spectrum, select the amplitude maximal value as resonance frequency (μ
m, ν
n), time obtain fundamental frequency (μ with resonance frequency divided by level
0, ν
0).
Wherein, m and n be x and y direction resonance frequency the level time.
E. by the size of the corresponding master grating of raster pitch and fundamental frequency computed image.
Particularly, utilize raster pitch (P
x, P
y) and fundamental frequency (μ
0, ν
0), according to the relation that fundamental frequency equates with the grid number of lines, adopt the size (l of the corresponding master grating of formula (3) computed image
x, l
y).
(l
x,l
y)=(P
xμ
0,P
yν
0) （3）
F. demarcate microscopical enlargement factor by the ratio of the size of the display size of image and the corresponding master grating of image.
Particularly, utilize the display size (I of image
x, I
y) and the size (l of the corresponding master grating of image
x, l
y), adopt formula (4) to demarcate microscopical enlargement factor (MAG
x, MAG
y).
In a preferred embodiment of the invention, the preferential level time high resonance frequency of selecting among the step C.Inferior select high more of the level of resonance frequency, the precision that enlargement factor is demarcated is high more.
Need to prove that grating above can be orthogonal grating or unidirectional grating.When grating is orthogonal grating, can demarcate two enlargement factors on the mutually perpendicular direction simultaneously, demarcate the efficient height.When grating is unidirectional grating, execution in step A-F on grating principal direction, the enlargement factor of demarcation grating principal direction correspondence; With 90 ° of grating rotating, repeating step A-F on grating principal direction again is to demarcate the enlargement factor of other direction.
For making those skilled in the art understand the present invention better, it is as follows to enumerate a specific embodiment again.
The microscope of being demarcated is FEI SEM quanta450, is reference with the master grating of 1000 lines per millimeters, as shown in Figure 2, is placed in the microscope stage, and it is clear to imaging to adjust focal length, is 10000 * images acquired in the enlargement factor that shows.The image of gathering is carried out Fast Fourier Transform (FFT), obtain spatial frequency spectrum.Selecting level inferior in spatial frequency spectrum is between the resonance frequency location of (10,10), as shown in Figure 3.Adopt formula (1) to carry out local high resolving power discrete Fourier transformation and obtain local high resolution space frequency spectrum.
μ=μ
d,μ
d+D,μ
d+2D,…,μ
u
ν=ν
d,ν
d+D,ν
d+2D,…,ν
u
Wherein, (μ ν) is local high resolution space frequency spectrum to F; M=1024 is the pixel count of image x direction, and N=884 is the pixel count of image y direction; (x is that (x y) locates gray-scale value to image coordinate y) to f; D=0.01 is the frequency increment of local high resolving power Fourier transform; μ is an x director space frequency, and ν is a y director space frequency; (μ
d, μ
u)=(299,301) be the frequency separation of the local high resolving power Fourier transform of image x direction, (ν
d, ν
u)=(258,260) be the frequency separation of the local high resolving power Fourier transform of image y direction; I is an imaginary unit; π is a circular constant; E is the natural logarithm truth of a matter;
In local high resolution space frequency spectrum, select the amplitude maximal value as resonance frequency (μ
m, ν
n)=(300.86,259.33), utilize formula (2) to calculate fundamental frequency (μ
0, ν
0)=(30.086,25.933).
Wherein, m=10 and n=10 be x and y direction resonance frequency the level time.
Utilize raster pitch (P
x, P
y)=(1um, 1um) and fundamental frequency (30.086,25.933), according to the relation that fundamental frequency equates with the grid number of lines, adopt the corresponding master grating of formula (3) computed image size (30.086um, 25.933um).
(l
x,l
y)=(P
xμ
0,P
yν
0) （3）
Utilize image display size (298500um, 257686um) and the size of the corresponding master grating of image (30.086um 25.933um), adopts formula (4) to demarcate microscopical enlargement factor (MAG
x, MAG
y)=(9921.5,9936.7).
The present invention utilizes Fourier transform to demarcate microscopical enlargement factor, the precision height; Error when directly determining fundamental frequency adopts and determines senior time resonance frequency, utilizes the method for the relation calculating fundamental frequency of resonance frequency and fundamental frequency again; In addition, determine to dwindle between the inferior resonance frequency location of certain one-level the computation interval of local high resolving power discrete Fourier transformation by the method for utilizing Fast Fourier Transform (FFT) earlier, and then reduce computing time; This method calibration range is big, and applicability is wide.
In the description of this instructions, concrete feature, structure, material or characteristics that the description of reference term " embodiment ", " some embodiment ", " example ", " concrete example " or " some examples " etc. means in conjunction with this embodiment or example description are contained at least one embodiment of the present invention or the example.In this manual, the schematic statement to above-mentioned term not necessarily refers to identical embodiment or example.And concrete feature, structure, material or the characteristics of description can be with the suitable manner combination in any one or more embodiment or example.
Although illustrated and described embodiments of the invention above, be understandable that, the foregoing description is exemplary, can not be interpreted as limitation of the present invention, those of ordinary skill in the art can change the foregoing description under the situation that does not break away from principle of the present invention and aim within the scope of the invention, modification, replacement and modification.
Claims (7)
1. a microscope enlargement factor scaling method is characterized in that, comprises the steps:
A. the master grating with known raster pitch is reference, gathers raster image in the microscope that need are demarcated;
B. the image of gathering is carried out Fast Fourier Transform (FFT), obtain spatial frequency spectrum;
C. in spatial frequency spectrum, select to carry out local high resolving power discrete Fourier transformation and obtain local high resolution space frequency spectrum between the inferior resonance frequency location of certain one-level;
D. the frequency of selecting the amplitude maximum in local high resolution space frequency spectrum is as resonance frequency, with resonance frequency divided by the inferior fundamental frequency that obtains of level;
E. by the size of the corresponding master grating of raster pitch and fundamental frequency computed image;
F. demarcate microscopical enlargement factor by the ratio of the size of the display size of image and the corresponding master grating of image.
2. microscope enlargement factor scaling method as claimed in claim 1 is characterized in that, adopts following formula to carry out local high resolving power discrete Fourier transformation and obtains local high resolution space frequency spectrum:
μ=μ
d,μ
d+D,μ
d+2D,…,μ
u
ν=ν
d,ν
d+D,ν
d+2D,…,ν
u
Wherein, (μ ν) is local high resolution space frequency spectrum to F; M is the pixel count of image x direction, and N is the pixel count of image y direction; (x is that (x y) locates gray-scale value to image coordinate y) to f; D is the frequency increment of local high resolving power Fourier transform, and the size of D requires to adjust according to computational accuracy; μ is an x director space frequency, and ν is a y director space frequency; (μ
d, μ
u) be the frequency separation of the local high resolving power Fourier transform of image x direction, (ν
d, ν
u) be the frequency separation of the local high resolving power Fourier transform of image y direction; I is an imaginary unit; π is a circular constant; E is the natural logarithm truth of a matter.
3. microscope enlargement factor scaling method as claimed in claim 1 is characterized in that, the frequency that is defined in selection amplitude maximum in the local high resolution space frequency spectrum is as resonance frequency (μ
m, ν
n), time obtain fundamental frequency (μ with resonance frequency divided by level
0, ν
0), promptly
Wherein, m and n be x and y direction resonance frequency the level time.
4. microscope enlargement factor scaling method as claimed in claim 1 is characterized in that the definition raster pitch is (P
x, P
y) and fundamental frequency be (μ
0, ν
0), by the size (l of the corresponding master grating of following formula computed image
x, l
y): (l
x, l
y)=(P
xμ
0, P
yν
0).
5. microscope enlargement factor scaling method as claimed in claim 1 is characterized in that, utilizes the display size (I of image
x, I
y) and the size (l of the corresponding master grating of image
x, l
y), demarcate microscopical enlargement factor (MAG by following formula
x, MAG
y):
6. microscope enlargement factor scaling method as claimed in claim 1 is characterized in that, among the described step C, selects level time high resonance frequency.
7. microscope enlargement factor scaling method as claimed in claim 1 is characterized in that, described grating is orthogonal grating or unidirectional grating.
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|---|---|---|---|
| CN201310175694.8A CN103217275B (en) | 2013-05-14 | 2013-05-14 | Magnification factor calibration method for microscope |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310175694.8A CN103217275B (en) | 2013-05-14 | 2013-05-14 | Magnification factor calibration method for microscope |
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| CN103217275B CN103217275B (en) | 2015-04-15 |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104406519A (en) * | 2014-11-25 | 2015-03-11 | 深圳市计量质量检测研究院 | Method and device for digitally measuring magnification multiplying power |
| CN110579340A (en) * | 2019-09-17 | 2019-12-17 | 长江存储科技有限责任公司 | Method for judging magnification of transmission electron microscope and method for calibrating magnification of transmission electron microscope |
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| US6807314B1 (en) * | 1999-07-02 | 2004-10-19 | General Phosphorix, Llc | Method of precision calibration of a microscope and the like |
| US20050220362A1 (en) * | 1999-07-02 | 2005-10-06 | Arkady Nikitin | Method of precision calibration of a microscope and the like |
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2013
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Patent Citations (5)
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|---|---|---|---|---|
| US4818873A (en) * | 1987-10-30 | 1989-04-04 | Vickers Instruments (Canada) Inc. | Apparatus for automatically controlling the magnification factor of a scanning electron microscope |
| US5581631A (en) * | 1994-09-20 | 1996-12-03 | Neopath, Inc. | Cytological system image collection integrity checking apparatus |
| US6807314B1 (en) * | 1999-07-02 | 2004-10-19 | General Phosphorix, Llc | Method of precision calibration of a microscope and the like |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104406519A (en) * | 2014-11-25 | 2015-03-11 | 深圳市计量质量检测研究院 | Method and device for digitally measuring magnification multiplying power |
| CN110579340A (en) * | 2019-09-17 | 2019-12-17 | 长江存储科技有限责任公司 | Method for judging magnification of transmission electron microscope and method for calibrating magnification of transmission electron microscope |
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| CN103217275B (en) | 2015-04-15 |
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