CN113048898A - Nonlinear fluorescent dyeing film thickness measurement calibration method - Google Patents
Nonlinear fluorescent dyeing film thickness measurement calibration method Download PDFInfo
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- CN113048898A CN113048898A CN202110431140.4A CN202110431140A CN113048898A CN 113048898 A CN113048898 A CN 113048898A CN 202110431140 A CN202110431140 A CN 202110431140A CN 113048898 A CN113048898 A CN 113048898A
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- G01—MEASURING; TESTING
- G01B—MEASURING LENGTH, THICKNESS OR SIMILAR LINEAR DIMENSIONS; MEASURING ANGLES; MEASURING AREAS; MEASURING IRREGULARITIES OF SURFACES OR CONTOURS
- G01B11/00—Measuring arrangements characterised by the use of optical techniques
- G01B11/02—Measuring arrangements characterised by the use of optical techniques for measuring length, width or thickness
- G01B11/06—Measuring arrangements characterised by the use of optical techniques for measuring length, width or thickness for measuring thickness ; e.g. of sheet material
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Abstract
The invention provides a non-linear fluorescent dyeing film thickness measurement calibration method, which can effectively improve the measurement precision and the measurement range of the thickness of a fluorescent dyeing film. The present invention considers the nonlinearity between the fluorescence intensity and the thickness of the fluorescence-dyed film in an excitation-induced fluorescence imaging system. By using the fluorescent photographs of films with different standard thicknesses, a relation between the film thickness and the fluorescence intensity of each pixel unit is established based on a polynomial, and then a calibration coefficient equation set is constructed and solved to obtain a calibration coefficient table. And (3) shooting a fluorescence photo of the film to be detected, and calculating to obtain the film thickness corresponding to each pixel unit based on the calibration coefficient table and the relation between the film thickness and the fluorescence intensity. The measurement error caused by the nonlinearity between the fluorescence intensity and the thickness of the fluorescent dyeing film can be obviously reduced, and the measurement precision and the measuring range of the thickness of the fluorescent dyeing film can be effectively improved.
Description
Technical Field
The invention relates to the technical field of measurement and calibration, in particular to a nonlinear fluorescence staining film thickness measurement and calibration method.
Background
The thickness measuring technology of the fluorescent dyeing film is a technology for measuring the thickness of a transparent fluorescent medium film by exciting and inducing fluorescence imaging, can be used for measuring the thickness of liquid and solid films, such as the height of a structural gap, the depth of a surface groove, the thickness of an LED fluorescent film and the like, and is applied to the industries of machinery, electronics, biology and the like. According to the Beer-Lambert law, when a transparent liquid or solid film uniformly containing a fluorescent substance is excited by a normally incident light, the intensity of the generated fluorescence is related to the thickness of the film as follows:
the conventional calibration method considers that the fluorescence intensity and the film thickness are in a linear relation in a certain range, obtains a linear coefficient through experimental fitting, and directly calculates the film thickness through the fluorescence intensity. However, since the true relationship is non-linear, the linearization process can introduce truncation errors. Therefore, as the thickness of the film increases, the error between the measured value and the real thickness becomes larger, so that the measuring range based on the conventional calibration method is limited, and the precision is difficult to guarantee.
Disclosure of Invention
In view of this, the invention provides a method for measuring and calibrating the thickness of a fluorescent dyed film by taking nonlinear measurement into consideration, which can effectively improve the measurement precision and the measurement range of the thickness of the fluorescent dyed film.
The invention relates to a non-linear considered fluorescent dyeing film thickness measurement calibration method, which comprises the steps of using fluorescent photographs of films with different standard thicknesses, establishing a relational expression between the fluorescent intensity corresponding to each pixel unit and the film thickness based on a polynomial, constructing a calibration coefficient equation set according to the relational expression between the fluorescent intensity and the film thickness, and solving to obtain a calibration coefficient table; and calculating the film thickness corresponding to each pixel unit for the fluorescence photo of the film to be detected based on the calibration coefficient table and the relation between the film thickness and the fluorescence intensity.
Wherein the established relation between the thickness of the film and the fluorescence intensity is as follows:
wherein the content of the first and second substances,
the film thicknesses corresponding to the pixel units in the ith row and the jth column of the kth picture are obtained;
the calibration coefficients are n-order calibration coefficients corresponding to the ith row and the jth column of pixel units;
the fluorescence intensity of the kth picture corresponding to the ith row and jth column pixel units; i. j is the serial number of the row and the column where the pixel unit is positioned respectively; n is the order of the basis function; n is the total order of the basis function; k is the picture number, and K is 1, 2,3 … K; k is the total number of pictures, and K is N + 1.
Wherein, the calibration coefficient linear equation set is as follows:
wherein, the film thickness of the pixel unit to be measured is:
wherein the content of the first and second substances,
the film thicknesses corresponding to the pixel units in the ith row and the jth column of the kth picture are obtained;
the calibration coefficients are n-order calibration coefficients corresponding to the ith row and the jth column of pixel units;
the fluorescence intensity of the kth picture corresponding to the ith row and jth column pixel units; i. j is the serial number of the row and the column where the pixel unit is positioned respectively; n is the order of the basis function; n is the total order of the basis function; k is the picture number, and K is 1, 2,3 … K; k is the total number of pictures, and K is N + 1.
The method is realized by the following steps:
step S1, giving a polynomial order N to be considered;
step S2, taking fluorescent pictures of films with different standard thicknesses, wherein the total number is N + 1;
step S3, establishing a relation between the film thickness and the fluorescence intensity for a certain pixel unit;
constructing a calibration coefficient linear equation set according to the above;
s4, solving the calibration coefficient linear equation set obtained in the step S3 to obtain a calibration coefficient array;
step S5, executing steps S3-S4 to all pixel units to obtain a calibration coefficient table;
step S6, using the same optical system to shoot the fluorescence photo of the film to be measured;
and step S7, calculating the film thickness corresponding to each pixel unit according to the relation between the film thickness and the fluorescence intensity by using the calibration coefficient table obtained in the step S4.
The method is used for calibrating when the thickness of the liquid or solid film subjected to fluorescent staining is measured by a fluorescent imaging system.
In taking a fluorescent photograph of a film, the film is strictly focused on a film reference surface.
Has the advantages that:
the present invention considers the nonlinearity between the fluorescence intensity and the thickness of the fluorescence-dyed film in an excitation-induced fluorescence imaging system. By using the fluorescent photographs of films with different standard thicknesses, a relation between the film thickness and the fluorescence intensity of each pixel unit is established based on a polynomial, and then a calibration coefficient equation set is constructed and solved to obtain a calibration coefficient table. And (3) shooting a fluorescence photo of the film to be detected, and calculating to obtain the film thickness corresponding to each pixel unit based on the calibration coefficient table and the relation between the film thickness and the fluorescence intensity. The measurement error caused by the nonlinearity between the fluorescence intensity and the thickness of the fluorescent dyeing film can be obviously reduced, and the measurement precision and the measuring range of the thickness of the fluorescent dyeing film can be effectively improved.
Drawings
FIG. 1 is a flow chart of the actual operation process of the present invention.
Detailed Description
The invention is described in detail below by way of example with reference to the accompanying drawings.
According to the Beer-Lambert law, when a transparent liquid or solid film uniformly containing a fluorescent substance is excited by a normally incident light, the intensity of the generated fluorescence is related to the thickness of the film as follows:
since the fluorescence intensity and the film thickness originally have a negative exponential relationship and are nonlinear, the linearization process inevitably brings truncation errors. Therefore, the invention considers the measurement error caused by the nonlinear relation between the fluorescence intensity and the film thickness, uses the fluorescence photos of films with different standard thicknesses, establishes the relation between the fluorescence intensity and the film thickness corresponding to each pixel unit based on the polynomial, constructs and solves the calibration coefficient equation set to obtain the calibration coefficient table; and calculating the film thickness corresponding to each pixel unit for the fluorescence photo of the film to be detected based on the calibration coefficient table and the relation between the film thickness and the fluorescence intensity. The invention is suitable for the calibration when the thickness of the liquid or solid film subjected to fluorescent staining is measured by adopting a fluorescent imaging system.
Wherein the established relation between the thickness of the film and the fluorescence intensity is as follows:
wherein the content of the first and second substances,
the film thicknesses corresponding to the pixel units in the ith row and the jth column of the kth picture are obtained;
the calibration coefficients are n-order calibration coefficients corresponding to the ith row and the jth column of pixel units;
the fluorescence intensity of the kth picture corresponding to the ith row and jth column pixel units; i. j is the serial number of the row and the column where the pixel unit is positioned respectively; n is the order of the basis function; n is the total order of the basis function; k is the picture number, and K is 1, 2,3 … K; k is the total number of pictures, and K is N + 1.
Accordingly, a calibration coefficient linear equation set is constructed as follows:
after the calibration coefficient linear equation set is solved, a calibration coefficient array is obtained, and the film thickness of the pixel unit to be measured can be calculated according to the following formula:
the calibration method of the invention has the flow as shown in figure 1, and is specifically realized by the following steps:
step S1, giving a polynomial order N to be considered;
step S2, taking fluorescent pictures of films with different standard thicknesses (taking care to focus strictly on the film reference surface), wherein the total number is N + 1;
step S3, for a certain pixel unit, establishing a relation between the film thickness and the fluorescence intensity:
accordingly, a calibration coefficient linear equation set can be constructed:
s4, solving the calibration coefficient linear equation set obtained in the step S3 to obtain a calibration coefficient array;
step S5, executing steps S3-S4 to all pixel units to obtain a calibration coefficient table;
step S6, using the same optical system to shoot the fluorescence photo of the film to be measured (focusing to the film reference surface strictly when shooting the fluorescence photo of the film);
and step S7, calculating the film thickness corresponding to each pixel unit according to the relation between the film thickness and the fluorescence intensity by using the calibration coefficient table obtained in the step S4:
in summary, the above description is only a preferred embodiment of the present invention, and is not intended to limit the scope of the present invention. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Claims (7)
1. A fluorescence staining film thickness measurement calibration method considering nonlinearity is characterized in that fluorescence photographs of films with different standard thicknesses are used, a relational expression between fluorescence intensity and film thickness corresponding to each pixel unit is established based on a polynomial, a calibration coefficient equation set is established according to the relational expression between the fluorescence intensity and the film thickness and is solved, and a calibration coefficient table is obtained; and calculating the film thickness corresponding to each pixel unit for the fluorescence photo of the film to be detected based on the calibration coefficient table and the relation between the film thickness and the fluorescence intensity.
2. The method for calibrating the measurement of the thickness of a fluorescence-dyed film considering nonlinearity according to claim 1, wherein the relationship between the thickness of the film and the fluorescence intensity is:
wherein the content of the first and second substances,
the film thicknesses corresponding to the pixel units in the ith row and the jth column of the kth picture are obtained;
the calibration coefficients are n-order calibration coefficients corresponding to the ith row and the jth column of pixel units;
the fluorescence intensity of the kth picture corresponding to the ith row and jth column pixel units; i. j is the serial number of the row and the column where the pixel unit is positioned respectively; n is the order of the basis function; n is the total order of the basis function; k is the picture number, and K is 1, 2,3 … K; k is the total number of pictures, and K is N + 1.
3. The method for calibrating fluorescence-dyed film thickness measurement considering nonlinearity according to claim 2, wherein the calibration coefficient linear equation is as follows:
4. the method for measuring and calibrating the thickness of the fluorescence-dyed film considering the nonlinearity of claim 1, wherein the film thickness of the pixel unit to be measured is:
wherein the content of the first and second substances,
the film thicknesses corresponding to the pixel units in the ith row and the jth column of the kth picture are obtained;
the calibration coefficients are n-order calibration coefficients corresponding to the ith row and the jth column of pixel units;
the fluorescence intensity of the kth picture corresponding to the ith row and jth column pixel units; i. j is the serial number of the row and the column where the pixel unit is positioned respectively; n is the order of the basis function; n is the total order of the basis function; k is the picture number, and K is 1, 2,3 … K; k is the total number of pictures, and K is N + 1.
5. The method for calibrating the measurement of the thickness of the fluorescence-dyed film considering the nonlinearity according to any one of claims 1 to 4, comprising the steps of:
step S1, giving a polynomial order N to be considered;
step S2, taking fluorescent pictures of films with different standard thicknesses, wherein the total number is N + 1;
step S3, establishing a relation between the film thickness and the fluorescence intensity for a certain pixel unit;
constructing a calibration coefficient linear equation set according to the above;
s4, solving the calibration coefficient linear equation set obtained in the step S3 to obtain a calibration coefficient array;
step S5, executing steps S3-S4 to all pixel units to obtain a calibration coefficient table;
step S6, using the same optical system to shoot the fluorescence photo of the film to be measured;
and step S7, calculating the film thickness corresponding to each pixel unit according to the relation between the film thickness and the fluorescence intensity by using the calibration coefficient table obtained in the step S4.
6. The non-linearity-considered fluorescence-dyed film thickness measurement calibration method as claimed in any one of claims 1 to 5, used for calibration when measuring the thickness of a fluorescence-dyed liquid or solid film using a fluorescence imaging system.
7. The method for calibrating the thickness measurement of a fluorescence-dyed film considering nonlinearity as claimed in claim 5, wherein the fluorescence photograph of the film is taken with the film being focused exactly on the reference plane of the film.
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