CN110702615B - Color correction method for color digital transmission microscope - Google Patents
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Abstract
The invention discloses a color correction method of a color digital transmission microscope, which utilizes a micro transmission color card to correct the color of a microscopic image acquired by the color digital transmission microscope, and because an illumination light source of the digital microscope is usually fixed, the chromaticity characterization of a digital camera under the fixed condition is particularly suitable for the color correction of the digital microscope, the invention utilizes a color block in the micro transmission color card as a training sample to establish a forward chromaticity characterization model of the color digital transmission microscope.
Description
Technical Field
The invention relates to a color correction method for a microscope, in particular to a color correction method for a color digital transmission microscope.
Background
In color digital imaging systems, there are increasing demands placed on the color quality of images. The colors of digital microscopic imaging systems are formed with the particularity that, on the one hand, they are adapted to the sensory properties of the human eye and, on the other hand, they reflect the primary colors of the object as realistically as possible. Most of the existing color digital transmission microscopes are applied to medical detection, and many pathological characteristics are distinguished by the color change of cells, so that the requirement on color authenticity is high. The microscopic image processing software is an integral part of a digital microscopic imaging system, converts data acquired by an image sensor into a color image consistent with human vision, and provides powerful image post-processing and analysis functions. However, the processing software has a very high requirement on the color quality of the image of the digital microscopic imaging system, and if the color quality of the image is not good, the visual effect is affected, and the post-processing and analysis of the image are also greatly affected, so that the color correction of the image of the color digital transmission microscope is required. At present, most of color correction solutions adopt a white balance algorithm, however, the white balance algorithm is only a rough color correction method, and accurate color correction of a digital microscopic imaging system cannot be realized.
Disclosure of Invention
The invention aims to provide a color correction method of a color digital transmission microscope, which can improve the color quality of an image.
The technical scheme adopted by the invention for solving the technical problems is as follows: a color correction method for a color digital transmission microscope comprises the following steps:
(1) the method comprises the steps that a color camera is used in a color digital transmission microscope to obtain RGB values of all color blocks in a micro transmission color card, and the RGB values are defined as an RGB matrix with the number of the color blocks on the micro transmission color card as the number of lines and the number of color camera channels as the number of columns, wherein each line represents an RGB 3 channel response value corresponding to one color block, and each column represents the response value of all the color blocks output by one channel;
(2) the method comprises the steps of continuously shooting after an illumination light source of a microscope is turned off, obtaining dark current response values of a color camera, defining the dark current response values as a dark current response matrix with the number of color blocks on a micro transmission color card as the number of lines and the number of color camera channels as the number of columns, wherein each line represents the dark current response value of 3 channels of RGB corresponding to one color block, then measuring XYZ tristimulus values of all color blocks in the micro transmission color card by using a spectrophotometer, and defining the dark current response values as a tristimulus matrix with the number of color blocks on the micro transmission color card as the number of lines and the number of color camera channels as the number of columns, wherein each line represents the XYZ tristimulus values corresponding to one color block;
(3) placing the glass sheet with the transmittance of 70% on an objective table, shooting RGB images of the glass sheet by using a color camera, and correcting illumination space nonuniformity of RGB response values to obtain a color block response matrix of the miniature transmission color card after the illumination space nonuniformity is corrected;
(4) carrying out nonlinear correction on RGB response values by utilizing Y values in tristimulus values of neutral color blocks in the miniature transmission color card and corresponding RGB values;
(5) establishing a forward chromaticity characterization model of the color digital transmission microscope, namely a conversion relation between RGB values and XYZ tristimulus values;
(6) shooting and collecting RGB images of a target object, deducting dark current pixel by pixel, and correcting illumination space nonuniformity and nonlinearity of RGB response values to obtain a corrected response value matrix of the target object;
(7) and calculating the sRGB response value of the target object by using the tristimulus value of the target object to obtain an sRGB image which is the microscopic image after color correction. Utilizing tristimulus values S of target objectstAnd calculating the sRGB response value, wherein the sRGB image is the microscopic image after color correction.
The specific color correction method comprises the following steps:
(1) turning on the lighting source of the color digital transmission microscope, preheating for a period of time, placing the micro transmission color card below the objective table eyepiece, shooting the micro transmission color card, obtaining RGB values of all color blocks in the color card image, and recording the RGB values as
P is a matrix with n rows and 3 columns, wherein n is the number of color blocks on the micro transmission color card, 3 is the number of color camera channels, each row represents the RGB 3 channel response value corresponding to one color block, and each column represents the response value of n color blocks output by one channel.
(2) Turning off the microscope illumination light source and using the color camera to shoot, obtaining the dark current response value of the color camera and recording the dark current response value
D is a matrix with n rows and 3 columns, wherein n is the number of color blocks on the micro transmission color card, each row represents the dark current response value of an RGB 3 channel corresponding to one color block, and the response value of the color block after subtracting the dark current is PDP-D, and then the XYZ tristimulus values of all color blocks in the miniature transmission color chart were measured using a spectrophotometer and recorded as
Expressing that S is a matrix with n rows and 3 columns, n is the number of color blocks on the miniature transmission color card, and each row expresses XYZ tristimulus values corresponding to one color block;
(3) placing the glass sheet with the transmittance of 70 percent on an object stage, shooting RGB images of the glass sheet by using a color camera, correcting illumination nonuniformity of color block response values in the micro transmission color card channel by using RGB values of the glass sheet,
wherein the content of the first and second substances,
the corrected response value of the kth channel of the ith color block in the color card, k belongs to { R, G, B },
the average response value of the k channel of the glass sheet image,
the response value of the kth channel of the glass sheet image corresponding to the ith color block in the micro transmission color card image,
obtaining color block response matrix of the color card after illumination space nonuniformity correction for response value of ith color block kth channel in the color card, and recording the matrix as
Represents PLThe matrix is a matrix with n rows and 3 columns, n is the number of color blocks on the miniature transmission color card, each row represents RGB value after color block correction, and each column represents the value under a certain color channel of all the color blocks;
(4) respectively establishing nonlinear correction relationship of RGB three channels by using Y value in tristimulus value of neutral color block in miniature transmission color card and its corresponding RGB value,
yi＝c1pik 3+c2pik 2+c3pik+c4
in the formula, yiNormalized Y stimulus value, p, representing the ith neutral color patch on the miniature transmission color chipikRepresents the normalized response value of the ith neutral patch at the kth channel, k ∈ { R, G, B }, c1，c2，c3，c4Fitting coefficients of cubic polynomials are respectively adopted, the values of the four fitting coefficients are respectively between 0 and 3 and cannot be simultaneously 0, and the RGB response values P of all color blocks in the micro transmission color card are corrected by utilizing the above formulaLAnd recording the corrected color block response value as
Is represented by PcA matrix of n rows and 3 columns;
(5) establishing a forward chromaticity characterization model of a color digital microscope, i.e. establishing a conversion relation between RGB value and XYZ tristimulus values, and using corrected color block response value PcAnd the tristimulus values S thereof, a conversion mathematical model between the tristimulus values S and the tristimulus values S is established, a conversion matrix Z between the tristimulus values S and the tristimulus values S is calculated by a least square method,
Z＝STPc(Pc TPc)-1
wherein, superscript T represents the transpose of the matrix, and superscript-1 represents the inverse of the matrix;
(6) using a color camera to capture and collect RGB images of a target object, subtracting dark current pixel by pixel and formulating
And formula yi＝c1pik 3+c2pik 2+c3pik+c4Respectively correcting the illumination space nonuniformity and nonlinearity of the RGB response value of the target object to obtain a corrected response value matrix of the target object, and recording the corrected response value matrix as
Represents PtFor a matrix of m rows and 3 columns, m being the number of pixels in the image, XYZ tristimulus values S of the target object are calculated using a conversion matrix Zt，
St＝ZPt；
(7) Utilizing tristimulus values S of target objectstCalculating its sRGB response value, sRGB ═ MStObtaining sRThe GB image is a microscopic image after color correction, wherein
Compared with the prior art, the method has the advantages that the color of the microscopic image acquired by the color digital transmission microscope is corrected by utilizing the miniature transmission color chip, the lighting source of the digital microscope is usually fixed, and the digital camera chromaticity characterization under the fixed condition is particularly suitable for the color correction of the digital microscope, so that the method utilizes the color block in the miniature transmission color chip as the training sample to establish the forward chromaticity characterization model of the color digital transmission microscope, can realize the accurate color correction of the shot target by utilizing the model, improves the color precision of the color digital transmission microscope, accords with the color perception of human eyes, realizes the what you see is what you get effect, obviously improves the identification degree of the target object in the microscopic detection, and realizes the accurate identification and positioning of the target object.
Drawings
FIG. 1 is a flow chart for correcting color of a color digital transmission microscope using the method of the present invention.
Detailed Description
The invention is described in further detail below with reference to the following examples of the drawings.
Taking a biological microscope as an example, a method for correcting the color of a color digital transmission microscope based on a micro transmission color chip is described, wherein the micro transmission color chip adopts a transmission color chip containing 45 color blocks, and the size of the color chip is 3mm multiplied by 4 mm.
The method for correcting the color of the color digital transmission microscope based on the micro transmission color card comprises the following steps:
(1) turning on the lighting source of the color digital transmission microscope, preheating for a period of time, placing the miniature transmission color card below the objective table eyepiece, shooting the miniature transmission color card, obtaining the RGB values of all color blocks in the color card image, and recording the RGB values as
P is a matrix of 45 rows and 3 columns, where 45 is a micro-scale transmission colorThe number of color blocks on the card, 3, is the number of color camera channels, each line represents the RGB three-channel response values corresponding to one color block, and each column represents the response values of 45 color blocks output by one channel.
(2) Turning off the microscope illumination light source and shooting to obtain the dark current response value of the color card
45 is the number of color blocks on the micro transmission color card, each row represents the dark current response value of the RGB 3 channel corresponding to one color block, and the response value of the color block after deducting the dark current is PDP-D. XYZ tristimulus values of all color blocks in a micro transmission color card were measured using a spectrophotometer CE7000A manufactured by Icely corporation
And 45 is the number of color blocks on the micro transmission color card, and each row represents XYZ tristimulus values corresponding to one color block.
(3) And carrying out illumination space nonuniformity correction on the RGB response values. Placing a glass sheet with the transmittance of 70 percent on an object stage, shooting RGB images of the glass sheet, correcting the illumination nonuniformity of color block response values in the color card channel by using the RGB values of the glass sheet, wherein the correction formula is as follows,
wherein the content of the first and second substances,
the average response value of the k (k epsilon { R, G, B }) channel of the glass sheet image,
the response value of the kth channel of the glass sheet image at the ith color block in the corresponding color card image,
the response value of the kth channel of the ith color block in the color card,
the corrected response value of the k channel of the ith color block in the color card. The color block response value matrix of the color card after the illumination space nonuniformity is corrected is PL。
(4) The RGB response values are non-linearly corrected. Respectively establishing RGB three-channel nonlinear correction relationship by using Y value in tristimulus value of neutral color block in the miniature transmission color card and its corresponding RGB value, as shown in the following formula,
yi＝c1pik 3+c2pik 2+c3pik+c4 (2)
in the above formula, yiNormalized Y stimulus value, p, representing the ith neutral color BlockikDenotes the normalized response value, c, of the ith neutral patch at the kth (k ∈ { R, G, B }) channel1，c2，c3，c4The fitting coefficients of the cubic polynomials are respectively, the values of the four fitting coefficients are respectively between 0 and 3, and cannot be simultaneously 0, in this embodiment, the values of the four fitting coefficients are respectively c 1-1.637, c 2-1.997, c 3-1.502, and c 4-0.136. Correction of RGB response P of all color blocks in miniature transmissive color card by using the above formulaLAnd the corrected color block response value is recorded as
(5) Establishing a forward chromaticity characterization model of the color digital microscope, namely establishing a conversion relation between RGB values and XYZ tristimulus values. Using the corrected color block response value PcAnd the tristimulus values S of the three-dimensional transformation matrix are used for establishing a transformation mathematical model between the three-dimensional transformation matrix and the tristimulus values S, and a transformation matrix Z between the three-dimensional transformation matrix and the tristimulus values S is calculated by a least square method, namely
Z＝STPc(Pc TPc)-1 (3)
Where superscript T represents the transpose of the matrix and superscript-1 represents the inverse of the matrix.
(6) Capturing and collecting RGB images of biological tissue slicesSubtracting dark current pixel by pixel and respectively correcting the illumination space nonuniformity and nonlinearity of the RGB response value of the biological tissue slice according to the formula (1) and the formula (2) to obtain a response value matrix after the biological tissue slice is corrected
m is the number of pixels in the image. Calculating XYZ tristimulus values S of the biological tissue section using the conversion matrix Z in equation (3)tI.e. by
St＝ZPt (4)
(7) Tristimulus value S using biological tissue slicestAnd calculating the sRGB response value, as shown in formula (5). The sRGB image is a microscopic image after color correction.
sRGB＝MSt (5)
Wherein the matrix
Claims (2)
1. A color correction method for a color digital transmission microscope is characterized by comprising the following steps:
(1) the method comprises the steps that a color camera is used in a color digital transmission microscope to obtain RGB values of all color blocks in a micro transmission color card, and the RGB values are defined as an RGB matrix with the number of the color blocks on the micro transmission color card as the number of lines and the number of color camera channels as the number of columns, wherein each line represents an RGB 3 channel response value corresponding to one color block, and each column represents the response value of all the color blocks output by one channel;
(2) the method comprises the steps of continuously shooting after an illumination light source of a microscope is turned off, obtaining dark current response values of a color camera, defining the dark current response values as a dark current response matrix with the number of color blocks on a micro transmission color card as the number of lines and the number of color camera channels as the number of columns, wherein each line represents the dark current response value of 3 channels of RGB corresponding to one color block, then measuring XYZ tristimulus values of all color blocks in the micro transmission color card by using a spectrophotometer, and defining the dark current response values as a tristimulus matrix with the number of color blocks on the micro transmission color card as the number of lines and the number of color camera channels as the number of columns, wherein each line represents the XYZ tristimulus values corresponding to one color block;
(3) placing the glass sheet with the transmittance of 70% on an objective table, shooting RGB images of the glass sheet by using a color camera, and correcting illumination space nonuniformity of RGB response values to obtain a color block response matrix of the miniature transmission color card after the illumination space nonuniformity is corrected;
(4) carrying out nonlinear correction on RGB response values by utilizing Y values in tristimulus values of neutral color blocks in the miniature transmission color card and corresponding RGB values;
(5) establishing a forward chromaticity characterization model of the color digital transmission microscope, namely a conversion relation between an RGB value and an XYZ tristimulus value;
(6) shooting and collecting RGB images of a target object, deducting dark current pixel by pixel, and correcting illumination space nonuniformity and nonlinearity of RGB response values to obtain a corrected response value matrix of the target object;
(7) and calculating the sRGB response value of the target object by using the tristimulus value of the target object to obtain an sRGB image which is the microscopic image after color correction.
2. The color correction method of a color digital transmission microscope according to claim 1, wherein the specific correction method is as follows:
(1) turning on the lighting source of the color digital transmission microscope, preheating for a period of time, placing the miniature transmission color card below the objective table eyepiece, shooting the miniature transmission color card, obtaining the RGB values of all color blocks in the color card image, and recording the RGB values as
P is a matrix with n rows and 3 columns, wherein n is the number of color blocks on the miniature transmission color card, 3 is the number of color camera channels, each row represents the RGB 3 channel response value corresponding to one color block, and each column represents the response value of n color blocks output by one channel;
(2) turning off the microscope illumination light source and using the color camera to shoot, obtaining the dark current response value of the color camera and recording the dark current response value
D is a matrix with n rows and 3 columns, wherein n is the number of color blocks on the micro transmission color card, each row represents the dark current response value of an RGB 3 channel corresponding to one color block, and the response value of the color block after subtracting the dark current is PDP-D, and then the XYZ tristimulus values of all color blocks in the miniature transmission color chart were measured using a spectrophotometer and recorded as
Expressing that S is a matrix with n rows and 3 columns, n is the number of color blocks on the miniature transmission color card, and each row expresses XYZ tristimulus values corresponding to one color block;
(3) placing the glass sheet with the transmittance of 70 percent on an object stage, shooting RGB images of the glass sheet by using a color camera, correcting illumination nonuniformity of color block response values in the micro transmission color card channel by using RGB values of the glass sheet,
wherein, the first and the second end of the pipe are connected with each other,
the corrected response value of the kth channel of the ith color block in the color card, k belongs to { R, G, B },
the average response value of the k channel of the glass sheet image,
the response value of the kth channel of the glass sheet image corresponding to the ith color block in the micro transmission color card image,
obtaining color block response moment of the color card after correcting the illumination space nonuniformity for the response value of the kth channel of the ith color block in the color cardArray, as
Represents PLThe matrix is a matrix with n rows and 3 columns, n is the number of color blocks on the miniature transmission color card, each row represents RGB value after color block correction, and each column represents the value under a certain color channel of all the color blocks;
(4) respectively establishing nonlinear correction relationship of RGB three channels by using Y value in tristimulus value of neutral color block in miniature transmission color card and its corresponding RGB value,
yi＝c1pik 3+c2pik 2+c3pik+c4
in the formula, yiNormalized Y stimulus value, p, representing the ith neutral color patch on the miniature transmission color chipikRepresents the normalized response value of the ith neutral patch at the kth channel, k ∈ { R, G, B }, c1，c2，c3，c4Fitting coefficients of cubic polynomials are respectively adopted, the values of the four fitting coefficients are respectively between 0 and 3 and cannot be simultaneously 0, and the RGB response values P of all color blocks in the micro transmission color card are corrected by utilizing the above formulaLAnd recording the corrected color block response value as
Is represented by PcA matrix of n rows and 3 columns;
(5) establishing a forward chromaticity characterization model of a color digital microscope, i.e. establishing a conversion relation between RGB value and XYZ tristimulus values, and using corrected color block response value PcAnd the tristimulus values S thereof, a conversion mathematical model between the tristimulus values S and the tristimulus values S is established, a conversion matrix Z between the tristimulus values S and the tristimulus values S is calculated by a least square method,
Z＝STPc(Pc TPc)-1
wherein, superscript T represents the transpose of the matrix, and superscript-1 represents the inverse of the matrix;
(6) capturing and acquiring RGB images of a target object pixel by pixel using a color cameraDeducting dark current according to formula
And formula yi＝c1pik 3+c2pik 2+c3pik+c4Respectively correcting the illumination space nonuniformity and nonlinearity of the RGB response value of the target object to obtain a corrected response value matrix of the target object, and recording the corrected response value matrix as
Represents PtFor a matrix of m rows and 3 columns, m being the number of pixels in the image, XYZ tristimulus values S of the target object are calculated using a conversion matrix Zt，
St＝ZPt；
(7) Utilizing tristimulus values S of a target objecttCalculating the sRGB response value, wherein sRGB is MStObtaining an sRGB image which is a microscopic image after color correction, wherein
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