CN107770434B - Rapid focusing adjustment method - Google Patents
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Abstract
The invention relates to the field of image shooting and provides a quick focusing adjustment method. The method comprises a preparation step and a monitoring step, wherein the monitoring step comprises a photographing step, a judging step and a focusing step, the first definition value, the second definition value and the distance between the photographing positions corresponding to the first definition value and the second definition value are obtained through the preparation step, then the distance between the photographing module and the platform where the photographing object is located is rapidly predicted and focused according to the first distance = | (the first definition value-the third definition value) ÷ (the first definition value-the second definition value) | × A in the monitoring step, the distance between the photographing module and the photographing object required to move relative to the platform where the photographing object is located is always the same as or similar to the optimal focusing distance, and the picture photographed by the photographing module is always clear.
Description
Technical Field
The invention relates to the field of image shooting, in particular to a quick focusing adjustment method.
Background
In the prior art, a shooting module is often moved by a fixed step length and shoots a shooting object, so as to obtain a series of images with different focal lengths, then the images with the best definition are obtained by analyzing the definition values of the images, the position corresponding to the image with the best definition is defined as an optimal focusing position, and then the shooting module is moved to the optimal focusing position for shooting. However, after the focus adjustment is completed, if the distance between the photographing module and the photographing object is changed, the above steps are often repeated to obtain the optimal focus position, and the focus adjustment speed is too slow.
A new method of enabling fast focus adjustment is therefore needed.
Disclosure of Invention
The invention aims to provide a quick focusing adjustment method, and aims to solve the problem of low focusing adjustment speed in the prior art.
In order to achieve the object of the invention, the invention provides a fast focusing adjustment method, which comprises the following steps:
the method comprises the following steps that a control module controls a shooting module to respectively shoot a shooting object at an optimal focusing position and a preset position to obtain a first image and a second image, and then the control module respectively calculates the first image and the second image by using a definition value calculation method to obtain a corresponding first definition value and a corresponding second definition value; the control module controls the shooting module to return to the optimal focusing position; the optimal focusing position and the preset position are both positioned on a Z axis vertical to the plane of the shooting object, and the distance between the optimal focusing position and the preset position is A;
a monitoring step, wherein the monitoring step comprises a photographing step, a judging step and a focusing step;
in the step of photographing, the control module controls the photographing module to photograph to obtain a third image, and then the control module calculates the third image by using a definition value calculation method to obtain a third definition value;
in the judging step, if the difference value between the first definition value and the third definition value does not exceed the threshold range, focusing is finished; if the difference value between the first definition value and the third definition value exceeds the threshold value range, entering a focusing step;
in the focusing step, the control module controls a platform where the shooting object is located to move a first distance in a first direction and shoot to obtain a fourth image, and then the control module calculates the fourth image by using a definition value calculation method to obtain a fourth definition value; first spacing = | (first definition value-third definition value) ÷ (first definition value-second definition value) | × a;
comparing the fourth sharpness value with the third sharpness value;
if the fourth definition value is large, defining the fourth definition value as a third definition value, and entering a judging step;
if the fourth definition value is small, the control module controls the platform where the shooting object is located to move a second distance in a second direction and shoot to obtain a fifth image, and then the control module calculates the fifth image by using a definition value calculation method to obtain a fifth definition value; defining the fifth definition value as the third definition value and the second direction as the first direction, and entering the judging step;
the first direction and the second direction are both vertical to a platform where the shooting object is located;
the second pitch is greater than the first pitch.
Wherein, the definition value calculation method comprises the following steps:
calculating the gray difference value of each pixel point in a target area in the image;
reserving the part with large gray difference values in all pixel points in the target area in the image according to a preset proportion;
and calculating the average value of all the gray level difference values reserved according to the preset proportion, and defining the average value as the definition value of the image.
The gray difference value of each pixel point is the gray difference value between each pixel point and four neighborhood pixel points thereof, or the gray difference value between each pixel point and eight neighborhood pixel points thereof.
Wherein the preset proportion is between 0.001% and 1%.
Wherein the target area is a rectangular area in the middle of the image.
Wherein, the step of taking a picture specifically is: the control module controls the shooting module to shoot every preset time to obtain a third image, and then the control module calculates the third image by using a definition value calculation method to obtain a third definition value.
Wherein the preset time is 1-3 s.
Wherein the distance between the best focus position and the preset position is 2-6 μm.
Wherein the shooting object is a region in which a sequencing reaction occurs in the high-throughput gene sequencing reaction chamber.
Wherein the first spacing = | (first-third sharpness value) ÷ (first-second sharpness value) | × a × B; b is a focusing coefficient; if | (= first sharpness value-third sharpness value) ÷ (first sharpness value-second sharpness value) | > 1, said B = | (first sharpness value-third sharpness value) ÷ (first sharpness value-second sharpness value) |; if | is (first definition value-third definition value) ÷ (first definition value-second definition value) | ≦ 1, said B = 1.
And leveling the platform where the shooting object is positioned after the preparation step.
The preparation step further comprises a step of translating the platform where the shooting object is located relative to the shooting module, or the preparation step further comprises a step of translating the shooting module relative to the platform where the shooting object is located.
In order to achieve the object of the present invention, the present invention provides another fast focus adjustment method, comprising the steps of:
the method comprises the following steps that a control module controls a shooting module to respectively shoot a shooting object at an optimal focusing position and a preset position to obtain a first image and a second image, and then the control module respectively calculates the first image and the second image by using a definition value calculation method to obtain a corresponding first definition value and a corresponding second definition value; the control module controls the shooting module to return to the optimal focusing position; the optimal focusing position and the preset position are both positioned on a Z axis vertical to the plane of the shooting object, and the distance between the optimal focusing position and the preset position is A;
a monitoring step, wherein the monitoring step comprises a photographing step, a judging step and a focusing step;
in the step of photographing, the control module controls the photographing module to photograph to obtain a third image, and then the control module calculates the third image by using a definition value calculation method to obtain a third definition value;
in the judging step, if the difference value between the first definition value and the third definition value does not exceed the threshold range, focusing is finished; if the difference value between the first definition value and the third definition value exceeds the threshold value range, entering a focusing step;
in the focusing step, the control module controls the shooting module to move the first distance to the third direction and shoot to obtain a fourth image, and then the control module calculates the fourth image by using a definition value calculation method to obtain a fourth definition value; first spacing = | (first definition value-third definition value) ÷ (first definition value-second definition value) | × a;
comparing the fourth sharpness value with the third sharpness value;
if the fourth definition value is large, defining the fourth definition value as a third definition value, and entering a judging step;
if the fourth definition value is small, the control module controls the platform where the shooting object is located to move a second distance in the third direction and shoot to obtain a fifth image, and then the control module calculates the fifth image by using a definition value calculation method to obtain a fifth definition value; defining the fifth definition value as the third definition value, and entering a judging step;
the third direction is vertical to the platform where the shooting object is located;
the second pitch is greater than the first pitch.
In order to achieve the object of the present invention, the present invention further provides a fast focus adjustment method, comprising the steps of:
the method comprises the following steps that a control module controls a shooting module to respectively shoot a shooting object at an optimal focusing position and a preset position to obtain a first image and a second image, and then the control module respectively calculates the first image and the second image by using a definition value calculation method to obtain a corresponding first definition value and a corresponding second definition value; the control module controls the shooting module to return to the optimal focusing position; the optimal focusing position and the preset position are both positioned on a Z axis vertical to the plane of the shooting object, and the distance between the optimal focusing position and the preset position is A;
a monitoring step, wherein the monitoring step comprises a photographing step, a judging step and a focusing step;
in the step of photographing, the control module controls the photographing module to photograph to obtain a third image, and then the control module calculates the third image by using a definition value calculation method to obtain a third definition value;
in the judging step, if the difference value between the first definition value and the third definition value does not exceed the threshold range, focusing is finished; if the difference value between the first definition value and the third definition value exceeds the threshold value range, entering a focusing step;
in the focusing step, the control module controls the platform where the shooting object is located to move the first distance in the fourth direction and shoot to obtain a fourth image, and then the control module calculates the fourth image by using a definition value calculation method to obtain a fourth definition value; first spacing = | (first definition value-third definition value) ÷ (first definition value-second definition value) | × a;
comparing the fourth sharpness value with the third sharpness value;
if the fourth definition value is large, defining the fourth definition value as a third definition value, and entering a judging step;
if the fourth definition value is small, the control module controls the shooting module to move the second distance in the fourth direction and shoot to obtain a fifth image, and then the control module calculates the fifth image by using a definition value calculation method to obtain a fifth definition value; defining the fifth definition value as the third definition value, and entering a judging step;
the fourth direction is vertical to the platform where the shooting object is located;
the second pitch is greater than the first pitch.
In order to achieve the object of the present invention, the present invention further provides another fast focus adjustment method, comprising the steps of:
the method comprises the following steps that a control module controls a shooting module to respectively shoot a shooting object at an optimal focusing position and a preset position to obtain a first image and a second image, and then the control module respectively calculates the first image and the second image by using a definition value calculation method to obtain a corresponding first definition value and a corresponding second definition value; the control module controls the shooting module to return to the optimal focusing position; the optimal focusing position and the preset position are both positioned on a Z axis vertical to the plane of the shooting object, and the distance between the optimal focusing position and the preset position is A;
a monitoring step, wherein the monitoring step comprises a photographing step, a judging step and a focusing step;
in the step of photographing, the control module controls the photographing module to photograph to obtain a third image, and then the control module calculates the third image by using a definition value calculation method to obtain a third definition value;
in the judging step, if the difference value between the first definition value and the third definition value does not exceed the threshold range, focusing is finished; if the difference value between the first definition value and the third definition value exceeds the threshold value range, entering a focusing step;
in the focusing step, the control module controls the shooting module to move the first distance to the fifth direction and shoot to obtain a fourth image, and then the control module calculates the fourth image by using a definition value calculation method to obtain a fourth definition value; first spacing = | (first definition value-third definition value) ÷ (first definition value-second definition value) | × a;
comparing the fourth sharpness value with the third sharpness value;
if the fourth definition value is large, defining the fourth definition value as a third definition value, and entering a judging step;
if the fourth definition value is small, the control module controls the shooting module to move the second distance in the sixth direction and shoot to obtain a fifth image, and then the control module calculates the fifth image by using a definition value calculation method to obtain a fifth definition value; defining the fifth definition value as a third definition value, defining the sixth direction as a fifth direction, and entering a judging step;
the fifth direction and the sixth direction are both vertical to a platform where the shooting object is located;
the second pitch is greater than the first pitch.
Therefore, the first definition value and the second definition value obtained in the preparation step and the distance between the first definition value and the shooting position corresponding to the second definition value can be used for quickly predicting the distance, which is required by the shooting module to move relative to the platform where the shooting object is located, in the monitoring step and focusing the distance, so that the distance between the shooting module and the shooting object is always the same as or similar to the optimal focusing distance, and the picture shot by the shooting module is always clear.
Drawings
FIG. 1 is a diagram of a sharpness value and focus curve image.
Fig. 2 is a flowchart of a fast focus adjustment method according to a first embodiment of the present invention.
Fig. 3 is a flowchart illustrating a fast focus adjustment method according to a first embodiment of the present invention.
Fig. 4 is a flowchart illustrating a fast focus adjustment method according to a second embodiment of the present invention.
Fig. 5 is a flowchart illustrating a fast focus adjustment method according to a third embodiment of the present invention.
Fig. 6 is a flowchart illustrating a fast focus adjustment method according to a fourth embodiment of the present invention.
FIG. 7 is a flowchart illustrating a method for calculating sharpness values according to an embodiment of the present invention.
Fig. 8 is a flowchart of a sharpness value calculation method in a fifth embodiment of the present invention.
Fig. 9 is a schematic flow chart of a possible fast focus adjustment process according to an embodiment of the present invention.
Fig. 10 is a schematic diagram of another possible flow chart during a fast focus adjustment process in an embodiment of the invention.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is described in further detail below with reference to the accompanying drawings and embodiments.
Under different focal length conditions, the shooting module shoots the shot object, and then the pictures are subjected to definition value calculation to obtain corresponding definition values; then, taking the focal length as the abscissa and the sharpness value as the ordinate, a sharpness value and focal length curve image as shown in fig. 1 can be obtained. As can be seen from fig. 1, the sharpness value and the focal length have a similar linear relationship on the same side of the best focus position.
Based on fig. 1, the present invention provides a first embodiment of a fast focus adjustment method, as shown in fig. 2 and 3, including the following steps:
s21, a preparation step, in which the control module controls the shooting module to respectively shoot the shooting object at the optimal focusing position and the preset position to obtain a first image and a second image, and then the control module respectively calculates the first image and the second image by using a definition value calculation method to obtain a corresponding first definition value and a corresponding second definition value; the control module controls the shooting module to return to the optimal focusing position; the optimal focusing position and the preset position are both positioned on a Z axis vertical to the plane of the shooting object, and the distance between the optimal focusing position and the preset position is A;
s22, a monitoring step, wherein the monitoring step comprises the following steps:
s221, a photographing step, wherein the control module controls the photographing module to photograph to obtain a third image, and then the control module calculates the third image by using a definition value calculation method to obtain a third definition value;
s222, a judging step, namely finishing focusing if the difference value between the first definition value and the third definition value does not exceed the threshold range; if the difference value between the first definition value and the third definition value exceeds the threshold value range, the step S223 and the step S of focusing are carried out;
s223, a focusing step, wherein the control module controls a platform where the shooting object is located to move a first distance in a first direction and shoot to obtain a fourth image, and then the control module calculates the fourth image by using a definition value calculation method to obtain a fourth definition value; first spacing = | (first definition value-third definition value) ÷ (first definition value-second definition value) | × a;
comparing the fourth sharpness value with the third sharpness value;
if the fourth sharpness value is large, defining the fourth sharpness value as the third sharpness value, and entering S222 and judging;
if the fourth definition value is small, the control module controls the platform where the shooting object is located to move a second distance in a second direction and shoot to obtain a fifth image, and then the control module calculates the fifth image by using a definition value calculation method to obtain a fifth definition value; defining the fifth definition value as the third definition value and the second direction as the first direction, and entering the step S222 and the step of judgment;
the first direction and the second direction are both vertical to a platform where the shooting object is located;
in one embodiment, the second pitch is greater than the first pitch, and preferably, the second pitch is 2 times the first pitch.
It should be noted that the photographic subject may be a substantially stationary landscape, or may be an object that has a certain motion capability or can be moved, including living objects and inanimate objects; the invention is particularly suitable for the situation that the displacement between the shooting object and the shooting module can occur in a certain time period. In the preparation step, the shooting object and the shooting module are both static; in the monitoring step, the photographic subject and/or the photographic module may move. As shown in fig. 1, the first sharpness value corresponding to the best focus position is maximum, or the first sharpness value corresponding to the best focus position is in a continuous interval containing the maximum sharpness values. The threshold range may be determined as needed, and in this embodiment, the threshold range may be set to be ± 0.1 × the first definition value. In a preferred embodiment of the present invention, the threshold range may be set within ± 0.03 × the first resolution value.
In the embodiment, the first definition value and the second definition value obtained in the preparation step and the distance between the shooting positions corresponding to the first definition value and the second definition value can be used for quickly predicting the distance, which is required by the shooting module to move relative to the platform where the shooting object is located, in the monitoring step and focusing the distance, so that the picture shot by the shooting module is always clear, and by using the method of the embodiment, the quick focusing adjustment on a possible moving object in a certain time period can be realized without perfect focusing equipment (PFS).
Based on fig. 1, a second embodiment of the present invention is further proposed, as shown in fig. 4, a fast focus adjustment method, including the following steps:
s41, a preparation step, in which the control module controls the shooting module to respectively shoot the shooting object at the optimal focusing position and the preset position to obtain a first image and a second image, and then the control module respectively calculates the first image and the second image by using a definition value calculation method to obtain a corresponding first definition value and a corresponding second definition value; the control module controls the shooting module to return to the optimal focusing position; the optimal focusing position and the preset position are both positioned on a Z axis vertical to the plane of the shooting object, and the distance between the optimal focusing position and the preset position is A;
s42, a monitoring step, wherein the monitoring step comprises a photographing step, a judging step and a focusing step;
s421, a photographing step, in which the control module controls the photographing module to photograph to obtain a third image, and then the control module calculates the third image by using a definition value calculation method to obtain a third definition value;
s422, a judging step, namely finishing focusing if the difference value between the first definition value and the third definition value does not exceed the threshold range; if the difference value between the first definition value and the third definition value exceeds the threshold value range, the step of S423 and focusing is carried out;
s423, focusing, wherein the control module controls the shooting module to move the first distance to the third direction and shoot to obtain a fourth image, and then the control module calculates the fourth image by using a definition value calculation method to obtain a fourth definition value; first spacing = | (first definition value-third definition value) ÷ (first definition value-second definition value) | × a;
comparing the fourth sharpness value with the third sharpness value;
if the fourth definition value is large, defining the fourth definition value as the third definition value, and entering the step S422 for judgment;
if the fourth definition value is small, the control module controls the platform where the shooting object is located to move a second distance in the third direction and shoot to obtain a fifth image, and then the control module calculates the fifth image by using a definition value calculation method to obtain a fifth definition value; defining the fifth definition value as the third definition value, and entering the step S422 for judgment;
the third direction is vertical to the platform where the shooting object is located;
the second pitch is greater than the first pitch.
Based on fig. 1, the present invention further provides a third embodiment, as shown in fig. 5, a fast focus adjustment method, including the following steps:
s51, a preparation step, in which the control module controls the shooting module to respectively shoot the shooting object at the optimal focusing position and the preset position to obtain a first image and a second image, and then the control module respectively calculates the first image and the second image by using a definition value calculation method to obtain a corresponding first definition value and a corresponding second definition value; the control module controls the shooting module to return to the optimal focusing position; the optimal focusing position and the preset position are both positioned on a Z axis vertical to the plane of the shooting object, and the distance between the optimal focusing position and the preset position is A;
s52, a monitoring step, wherein the monitoring step comprises a photographing step, a judging step and a focusing step;
s521, a photographing step, in which the control module controls the photographing module to photograph to obtain a third image, and then the control module calculates the third image by using a definition value calculation method to obtain a third definition value;
s522, a judging step, namely finishing focusing if the difference value between the first definition value and the third definition value does not exceed the threshold range; if the difference value between the first definition value and the third definition value exceeds the threshold range, the step S523 and the step of focusing are carried out;
s523, a focusing step, in which the control module controls the platform where the shooting object is located to move the first distance in the fourth direction and shoot to obtain a fourth image, and then the control module calculates the fourth image by using a definition value calculation method to obtain a fourth definition value; first spacing = | (first definition value-third definition value) ÷ (first definition value-second definition value) | × a;
comparing the fourth sharpness value with the third sharpness value;
if the fourth sharpness value is large, defining the fourth sharpness value as the third sharpness value, and entering the step S522 for judgment;
if the fourth definition value is small, the control module controls the shooting module to move the second distance in the fourth direction and shoot to obtain a fifth image, and then the control module calculates the fifth image by using a definition value calculation method to obtain a fifth definition value; defining the fifth definition value as the third definition value, and proceeding to S522 and the determining step;
the fourth direction is vertical to the platform where the shooting object is located;
the second pitch is greater than the first pitch.
Based on fig. 1, a fourth embodiment of the present invention is further proposed, as shown in fig. 6, a fast focus adjustment method, including the following steps:
s61, a preparation step, in which the control module controls the shooting module to respectively shoot the shooting object at the optimal focusing position and the preset position to obtain a first image and a second image, and then the control module respectively calculates the first image and the second image by using a definition value calculation method to obtain a corresponding first definition value and a corresponding second definition value; the control module controls the shooting module to return to the optimal focusing position; the optimal focusing position and the preset position are both positioned on a Z axis vertical to the plane of the shooting object, and the distance between the optimal focusing position and the preset position is A;
s62, a monitoring step, wherein the monitoring step comprises a photographing step, a judging step and a focusing step;
s621, a photographing step, wherein the control module controls the photographing module to photograph to obtain a third image, and then the control module calculates the third image by using a definition value calculation method to obtain a third definition value;
s622, in the judging step, if the difference value between the first definition value and the third definition value does not exceed the threshold range, focusing is finished; if the difference value between the first definition value and the third definition value exceeds the threshold range, the step S623 is carried out;
s623, focusing, wherein the control module controls the shooting module to move the first distance in the fifth direction and shoot to obtain a fourth image, and then the control module calculates the fourth image by using a definition value calculation method to obtain a fourth definition value; first spacing = | (first definition value-third definition value) ÷ (first definition value-second definition value) | × a;
comparing the fourth sharpness value with the third sharpness value;
if the fourth sharpness value is large, defining the fourth sharpness value as the third sharpness value, and entering the step S622 for judgment;
if the fourth definition value is small, the control module controls the shooting module to move the second distance in the sixth direction and shoot to obtain a fifth image, and then the control module calculates the fifth image by using a definition value calculation method to obtain a fifth definition value; defining the fifth definition value as a third definition value, defining the sixth direction as a fifth direction, and entering the step S622 and the step of judgment;
the fifth direction and the sixth direction are both vertical to a platform where the shooting object is located;
the second pitch is greater than the first pitch.
It should be noted that the first, second, third, and fourth embodiments are based on the same inventive concept, and only differ in that the control module controls the object to move the first and second pitches in the focusing step; when the object moved by the first distance is the same as the object moved by the second distance, the directions of the two movements are opposite; when the object moved by the first pitch and the object moved by the second pitch are different, the directions of the two movements are the same.
The method for calculating the sharpness value according to any of the above embodiments is not particularly limited, and will be further described with reference to a plurality of embodiments.
In one embodiment of the invention, the sharpness value calculation method is to calculate the sharpness value of the image by using the laplacian operator.
In an embodiment of the present invention, the laplacian operator Sum (LS) is a Sum of 8 neighborhood differential values of each pixel point in an image range, and the values thereof can also better reflect the change characteristic of the contrast of the image, and a larger value thereof indicates that the image is clearer.
Let g be an M × N image matrix, then the laplacian and expression are as follows:
in another embodiment of the invention, the sharpness value calculation method is to calculate the sharpness value of the image by using a Sober operator.
In another embodiment of the present invention, the sharpness value calculation method is to calculate the sharpness value of the image by using a gray-scale average gradient method.
The Gray Mean Gradient Method (GMG) is to square and then calculate the root Mean square of the difference of the Gray values of adjacent pixels in the length direction and the width direction of an image respectively, can better reflect the contrast change characteristics of the image, and the larger the value of the Gray Mean gradient method is, the clearer the image is.
Let g be an M × N image matrix, the gray scale average gradient value expression is as follows:
。
in another embodiment of the present invention, the method for calculating the sharpness value may refer to the method in CN201310424934.3, and as shown in fig. 7 in particular, the method includes the following steps:
s71, carrying out harmonic analysis processing on the image to obtain a harmonic analysis processing diagram corresponding to the image;
s72, fitting a closed curve corresponding to the image according to the texture features of the harmonic analysis processing image corresponding to the image;
s73, determining the area ratio between the graph corresponding to the closed curve and the corresponding image; the area ratio is defined as a sharpness value.
The fifth embodiment of the present invention is directed to the sharpness value calculation method, and as shown in fig. 8, the sharpness value calculation method includes the following steps:
s81, calculating the gray difference value of each pixel point in the target area in the image;
s82, reserving the parts with large gray difference values of all pixel points in the target area in the image according to a preset proportion;
and S83, calculating the average value of all the gray level difference values reserved according to the preset proportion, and defining the average value as the definition value of the image.
Compared with the definition value calculation method, the definition value calculation method in the implementation is simpler, quicker and more efficient, and the definition value calculation method of the embodiment can obtain better curve unimodal model of the definition value and the focal length and can finish focusing adjustment more quickly.
On the basis of the fifth embodiment, the present invention proposes a sixth embodiment, and the sharpness value calculation method includes the steps of:
s91, calculating the gray difference value of each pixel point in the target area in the image;
s92, reserving the parts with large gray difference values of all pixel points in the target area in the image according to a preset proportion;
s93, sorting the gray level difference values of all pixel points of the image, and then reserving the part with the larger gray level difference value according to a preset proportion;
and S64, calculating the average value of all the gray level difference values reserved according to the preset proportion, and defining the average value as the definition value of the image.
Compared with the fifth embodiment, this embodiment actually adds a sorting step, so that the subsequent steps of reservation and calculation are simpler.
In any of the above embodiments, the predetermined ratio may be set between 1% and 0.001%; in a preferred embodiment, the preset proportion is 0.01% of the total amount of the pixel points, and this embodiment not only considers the calculation amount of the monitoring step, but also ensures the representativeness of the obtained definition value.
For the gray scale difference of each pixel point, the gray scale difference of each pixel point may be a gray scale difference between each pixel point and two adjacent pixel points (e.g., two adjacent pixel points on the left side and the top side), may also be a gray scale difference between each pixel point and four adjacent pixel points (e.g., each adjacent pixel point on the top, bottom, left, right, and four sides), and may also be a gray scale difference between each pixel point and eight adjacent pixel points. In addition, it should be noted that, when the object to be photographed in the fifth embodiment and the sixth embodiment is a region in which a sequencing reaction occurs in a high-throughput gene sequencing reaction chamber, because the obtained image is a high-throughput gene sequencing image including a plurality of magnetic beads to be recognized, because the number of the magnetic beads is large and the length and width of the magnetic beads are generally only a few pixel points, the gray scale difference of the pixel points is screened by using a conventional edge detection method, which may result in a loss of more image details, especially details of the shapes of the magnetic beads, and the sharpness value of the obtained image is slightly poor in representation. By adopting the gray difference value calculation method and processing the gray difference values of all the pixel points in a screening mode, the maximum part of the gray difference values in each pixel point is kept and actually is the clearest part of the edge of the magnetic bead, the shape of a focal length curve of the average value of the gray difference values of a plurality of different images is considered to be in accordance with normal distribution, the definition recognition effect is very obvious, and the representativeness of the obtained definition value is better.
In a specific embodiment of the present invention, the gray scale difference of each pixel is preferably the gray scale difference between the pixel in its four neighboring domains, and the calculation formula of the gray scale difference between each pixel and the pixel in its four neighboring domains is:
wherein, S (x) is the gray scale difference of each pixel point, and f (x, y) is the gray scale value of the pixel point at the coordinate position (x, y). The scheme not only reflects more image information of each pixel point, improves the accuracy of the definition value calculation, but also simplifies the operation and gives consideration to the effect and the efficiency.
In an entire image, the focusing effect of the central region of the image is often optimal, and the sharpness of the region is representative of the sharpness of the entire image. In a preferred embodiment of the present invention, the target area is a rectangular area in the center of the image. According to the embodiment, the calculated amount in the definition value calculating process can be obviously reduced, and the efficiency is improved. When the object to be photographed is a region in which a sequencing reaction occurs in the high-throughput gene sequencing reaction chamber, the rectangular region in the middle of the image occupies one half to sixty-fourth, preferably one fourth, of the entire image area.
In view of the photographing steps in any of the above embodiments, the present invention provides a preferred embodiment, wherein the photographing step specifically includes: the control module controls the shooting module to shoot every preset time to obtain a third image, and then the control module calculates the third image by using a definition value calculation method to obtain a third definition value.
In this embodiment, the preset time may be set as needed, as long as the monitoring of the present invention can complete focusing within the preset time. The embodiment can effectively reduce the control frequency of the shooting module and the control module, reduce the calculation amount of the monitoring step and make the monitoring step more suitable for practical application. In a preferred embodiment of the present invention, the predetermined time is 1-3 s.
In any of the above embodiments, the distance a between the optimal focusing position and the preset position may be set according to specific situations. For example, when the subject is a region in which a sequencing reaction occurs in a high-throughput gene sequencing reaction chamber, the distance between the best focus position and the preset position is preferably 2 to 6 μm.
The invention is particularly suitable for the condition that the same shooting object needs to be shot for a long time, and the distance between the shooting object and the shooting module can be changed in the process, wherein the reason for the change can be active adjustment or objective condition change.
In an embodiment of the present invention, after the step of preparing, any one of the above fast focus adjustment methods further includes a step of leveling a platform on which the photographic subject is located. The leveling in this embodiment may be performed automatically by the instrument or manually.
In an embodiment of the present invention, the subject is a region in the high-throughput gene sequencing reaction chamber where a sequencing reaction occurs, and the preparation step further includes a leveling step before the sequencing and image-taking. Because high-throughput gene sequencing has a very high requirement on the precision of focusing adjustment, slight changes in shooting distance may cause the captured image to become blurred, thereby affecting subsequent data processing. In particular, in the leveling step, the platform on which the high-throughput gene sequencing reaction chamber is located needs to be leveled, which may cause the image shot by the shooting module which is already focused to be blurred and needs to be refocused and adjusted to make the shot image clear, and the speed of the focus adjustment in the process has a great influence on the time required for automatic leveling.
In another embodiment of the present invention, after the step of preparing, the method for adjusting fast focus further includes a step of translating a platform on which the photographic subject is located relative to the photographic module, or after the step of preparing, the method further includes a step of translating the photographic module relative to the platform on which the photographic subject is located. The translation refers to parallel movement, which may be parallel movement parallel to the ground, parallel movement perpendicular to the ground, or parallel movement at an angle to the ground.
In an embodiment of the present invention, the subject is a region in the high-throughput gene sequencing reaction chamber where a sequencing reaction occurs, and the preparing step further includes a sampling step in the sequencing sampling process. Because high-throughput gene sequencing has a very high requirement on the precision of focusing adjustment, slight changes in shooting distance may cause the captured image to become blurred, thereby affecting subsequent data processing.
Specifically, in the mapping step, since the photographing module can only photograph a part of the region where the sequencing reaction occurs in the high-throughput gene sequencing reaction chamber at a time, and the region where the sequencing reaction occurs may not be a completely flat plane as a whole, in order to ensure the definition of the image photographed in the whole sequencing reaction region, multiple focusing adjustments are required in the mapping process, and the speed of the focusing adjustments in the mapping process has a great influence on the time required for mapping.
In a specific embodiment of the invention, the method is applied to the field of high-throughput gene sequencing, the shooting object is an area where a sequencing reaction occurs in a high-throughput gene sequencing reaction chamber, the area is used for a leveling step before sequencing and image collection, and the first definition value corresponding to the optimal focusing position is the largest. Fig. 9 and 10 show 2 different cases that may occur in one fast focusing process, i.e., whether the direction of the first focusing (movement by the first pitch) is correct, respectively.
The preparation method comprises the following steps: the control module controls the shooting module to respectively shoot a shooting object at the optimal focusing position and the preset position to obtain a first image and a second image, then the control module respectively calculates the first image and the second image by using a definition value calculation method in the third embodiment to obtain a corresponding first definition value (dMax) and a corresponding second definition value (dRef), and the gray difference value of each pixel point is the gray difference value between the pixel points in the four adjacent domains; the control module controls the shooting module to return to the optimal focusing position; the best focus position and the preset position are both positioned on a Z axis vertical to the plane of the shooting object, and the distance between the best focus position and the preset position is 4 mu m.
Then the platform where the high-throughput gene sequencing reaction chamber is located starts the work of automatic leveling. At the same time, the monitoring step begins. The monitoring step comprises a photographing step, a judging step and a focusing step.
A photographing step: the control module controls the shooting module to shoot every 2s, an image obtained by each shooting is defined as a third image, and after the third image is obtained each time, the control module calculates the third image by using a definition value calculation method to obtain a third definition value (dTemp 1). The initial position corresponds to a distance (focal length) between the photographing module and the subject when the photographing module photographs for the first time in the photographing step. The first predicted position, the second predicted position, the third predicted position, and the fourth predicted position correspond to a distance (focal length) between the photographing module and the subject after the first movement, the second movement, the third movement, and the fourth movement, respectively.
A judging step: if the dMax-dTemp1 is less than or equal to 0.03 multiplied by dMax, the image shot by the current shooting module is considered to be clear, and focusing is finished; if the dMax-dTemp1 is more than 0.03 multiplied by dMax, the focusing step is carried out;
a focusing step: the control module controls the platform where the shooting object is located to move a first distance (D) in a first direction and shoot to obtain a fourth image, and then the control module calculates the fourth image by using a definition value calculation method to obtain a fourth definition value (dTemp 2); comparing the size of dTemp2 with dTemp 1; d = | (dMax-dTemp 1) × 4 ÷ (dMax-dRef) |; if dTemp2 is more than dTemp1, defining dTemp2 as dTemp1, and entering a judging step; if dEmp 2 is less than dEmp 1, it is indicated that the movement direction of the platform where the photographic object is located is wrong, the control module controls the platform where the photographic object is located to move 2D in the opposite direction of the first direction to obtain a fifth image, and then the control module calculates the fifth image by using a definition value calculation method to obtain a fifth definition value (dEmp 3); defining dTemp3 as dTemp1, and entering a judging step; until focusing is finished.
As shown in fig. 9, in the first focusing step, the moving direction of the stage on which the photographic subject is located is correct, and the subsequent 2 times of focusing, the moving direction of the stage on which the photographic subject is located is the same as the first time of focusing, and the focusing is completed after 3 times of focusing in total.
As shown in fig. 10, in the first focusing step, the first moving direction of the platform on which the shooting object is located is wrong, the second focusing needs to be performed, the platform is moved in the reverse direction for 2D (at this time, the moving direction is the correct direction), then after the shooting calculation, the judging step is performed again for judgment, and the subsequent third and fourth focusing are performed, wherein the moving direction of the platform on which the shooting object is located is the same as the second focusing direction, and the focusing is completed after 4 times of focusing in total. That is, when the control module controls the platform where the shooting object is located to move, the moving direction preferentially selects the direction which is determined to be the correct moving direction in the last focusing.
In addition, as shown in fig. 1, when the distance at which the subject and/or the photographing module may move is smaller, correspondingly, the linearity between the sharpness value and the interval in the focal length curve corresponding to the image captured in the monitoring step is better, and in this case, the required focusing time is smaller.
In order to improve the focusing efficiency of any of the above fast focus adjustment methods, the present invention proposes a seventh embodiment based on any of the above embodiments, which is an improvement on only the method for calculating the first spacing = | (first-third sharpness value) ÷ (first-second sharpness value) | × a × B; b is a focusing coefficient; if | (= first sharpness value-third sharpness value) ÷ (first sharpness value-second sharpness value) | > 1, said B = | (first sharpness value-third sharpness value) ÷ (first sharpness value-second sharpness value) |; if | is (first definition value-third definition value) ÷ (first definition value-second definition value) | ≦ 1, said B = 1.
As shown in fig. 1, the sharpness value maxima divide the sharpness value versus focus curve into two symmetrical or substantially symmetrical curves, and the linearity of the sharpness values is slightly different in different focus ranges. When | is between the first sharpness value and the second sharpness value | ≦ 1, it means that the third sharpness value is between the first sharpness value and the second sharpness value, and they are considered to be in the same linear interval, so that the focusing coefficient is defined as 1; when | is (first definition value-third definition value) ÷ (first definition value-second definition value) | > 1, the linear interval in which the third definition value is positioned is different from the linear interval in which the first definition value and the second definition value are positioned, and then the focusing coefficient is defined as | is (first definition value-third definition value) ÷ (first definition value-second definition value) |, the focusing efficiency can be effectively improved. In a preferred embodiment, the object to be photographed is a region in the high-throughput gene sequencing reaction chamber where a sequencing reaction occurs, and when the object to be photographed is used in combination with a leveling step before sequencing and image-taking or an image-taking step in a sequencing and image-taking process, focusing is often performed 3 to 4 times, or even only 1 time, for each movement of the photographing module or the platform where the object to be photographed is located, so that automatic focusing can be completed.
The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents and improvements made within the spirit and principle of the present invention are intended to be included within the scope of the present invention.
Claims (15)
1. A fast focus adjustment method, characterized in that the method comprises the steps of:
the method comprises the following steps that a control module controls a shooting module to respectively shoot a shooting object at an optimal focusing position and a preset position to obtain a first image and a second image, and then the control module respectively calculates the first image and the second image by using a definition value calculation method to obtain a corresponding first definition value and a corresponding second definition value; the control module controls the shooting module to return to the optimal focusing position; the optimal focusing position and the preset position are both positioned on a Z axis vertical to the plane of the shooting object, and the distance between the optimal focusing position and the preset position is A;
a monitoring step, wherein the monitoring step comprises a photographing step, a judging step and a focusing step;
in the step of photographing, the control module controls the photographing module to photograph to obtain a third image, and then the control module calculates the third image by using a definition value calculation method to obtain a third definition value;
in the judging step, if the difference value between the first definition value and the third definition value does not exceed the threshold range, focusing is finished; if the difference value between the first definition value and the third definition value exceeds the threshold value range, entering a focusing step;
in the focusing step, the control module controls a platform where the shooting object is located to move a first distance in a first direction and shoot to obtain a fourth image, and then the control module calculates the fourth image by using a definition value calculation method to obtain a fourth definition value; first distance | (first definition value-third definition value)/(first definition value-second definition value) | × a;
comparing the fourth sharpness value with the third sharpness value;
if the fourth definition value is large, defining the fourth definition value as a third definition value, and entering a judging step;
if the fourth definition value is small, the control module controls the platform where the shooting object is located to move a second distance in a second direction and shoot to obtain a fifth image, and then the control module calculates the fifth image by using a definition value calculation method to obtain a fifth definition value; defining the fifth definition value as the third definition value and the second direction as the first direction, and entering the judging step;
the first direction and the second direction are both vertical to a platform where a shooting object is located, and the first direction is opposite to the second direction;
the second pitch is greater than the first pitch.
2. The fast focus adjustment method according to claim 1, wherein the sharpness value calculation method comprises the steps of:
calculating the gray difference value of each pixel point in a target area in the image;
reserving the part with large gray difference values in all pixel points in the target area in the image according to a preset proportion;
and calculating the average value of all the gray level difference values reserved according to the preset proportion, and defining the average value as the definition value of the image.
3. The fast focus adjustment method of claim 2, wherein the gray scale difference of each pixel point is a gray scale difference between each pixel point and its four neighboring pixel points, or a gray scale difference between each pixel point and its eight neighboring pixel points.
4. The fast focus adjustment method of claim 2, wherein the predetermined ratio is between 0.001% and 1%.
5. The fast focus adjustment method of claim 2, wherein the target region is a rectangular region in the center of the image.
6. The fast focus adjustment method according to any one of claims 1 to 5, wherein the photographing step specifically comprises: the control module controls the shooting module to shoot every preset time to obtain a third image, and then the control module calculates the third image by using a definition value calculation method to obtain a third definition value.
7. The fast focus adjustment method of claim 6, wherein the predetermined time is 1-3 s.
8. The fast focus adjustment method of claim 6, wherein the distance between the best focus position and the preset position is 2-6 μm.
9. The fast focusing adjustment method according to any one of claims 1 to 5, wherein the subject is a region in which a sequencing reaction occurs in a high-throughput gene sequencing reaction chamber.
10. The fast focus adjustment method according to claim 1, wherein the first distance | (first definition value-third definition value) ÷ (first definition value-second definition value) | × a × B; b is a focusing coefficient; if | (first definition value-third definition value)/(first definition value-second definition value) | > 1, the B | (first definition value-third definition value)/(first definition value-second definition value)/; if | (first definition value-third definition value)/(first definition value-second definition value) | ≦ 1, the B ═ 1.
11. The fast focus adjustment method according to any one of claims 1 to 5, wherein the preparation step is followed by a step of leveling a platform on which the photographic subject is placed.
12. The fast focus adjustment method according to any one of claims 1 to 5, wherein the preparation step further comprises a step of translating a platform on which the photographic subject is placed with respect to the photographing module, or the preparation step further comprises a step of translating the photographing module with respect to the platform on which the photographic subject is placed.
13. A fast focus adjustment method, characterized in that the method comprises the steps of:
the method comprises the following steps that a control module controls a shooting module to respectively shoot a shooting object at an optimal focusing position and a preset position to obtain a first image and a second image, and then the control module respectively calculates the first image and the second image by using a definition value calculation method to obtain a corresponding first definition value and a corresponding second definition value; the control module controls the shooting module to return to the optimal focusing position; the optimal focusing position and the preset position are both positioned on a Z axis vertical to the plane of the shooting object, and the distance between the optimal focusing position and the preset position is A;
a monitoring step, wherein the monitoring step comprises a photographing step, a judging step and a focusing step;
in the step of photographing, the control module controls the photographing module to photograph to obtain a third image, and then the control module calculates the third image by using a definition value calculation method to obtain a third definition value;
in the judging step, if the difference value between the first definition value and the third definition value does not exceed the threshold range, focusing is finished; if the difference value between the first definition value and the third definition value exceeds the threshold value range, entering a focusing step;
in the focusing step, the control module controls the shooting module to move the first distance to the third direction and shoot to obtain a fourth image, and then the control module calculates the fourth image by using a definition value calculation method to obtain a fourth definition value; first distance | (first definition value-third definition value)/(first definition value-second definition value) | × a;
comparing the fourth sharpness value with the third sharpness value;
if the fourth definition value is large, defining the fourth definition value as a third definition value, and entering a judging step;
if the fourth definition value is small, the control module controls the platform where the shooting object is located to move a second distance in the third direction and shoot to obtain a fifth image, and then the control module calculates the fifth image by using a definition value calculation method to obtain a fifth definition value; defining the fifth definition value as the third definition value, and entering a judging step;
the third direction is vertical to the platform where the shooting object is located;
the second pitch is greater than the first pitch.
14. A fast focus adjustment method, characterized in that the method comprises the steps of:
the method comprises the following steps that a control module controls a shooting module to respectively shoot a shooting object at an optimal focusing position and a preset position to obtain a first image and a second image, and then the control module respectively calculates the first image and the second image by using a definition value calculation method to obtain a corresponding first definition value and a corresponding second definition value; the control module controls the shooting module to return to the optimal focusing position; the optimal focusing position and the preset position are both positioned on a Z axis vertical to the plane of the shooting object, and the distance between the optimal focusing position and the preset position is A;
a monitoring step, wherein the monitoring step comprises a photographing step, a judging step and a focusing step;
in the step of photographing, the control module controls the photographing module to photograph to obtain a third image, and then the control module calculates the third image by using a definition value calculation method to obtain a third definition value;
in the judging step, if the difference value between the first definition value and the third definition value does not exceed the threshold range, focusing is finished; if the difference value between the first definition value and the third definition value exceeds the threshold value range, entering a focusing step;
in the focusing step, the control module controls the platform where the shooting object is located to move the first distance in the fourth direction and shoot to obtain a fourth image, and then the control module calculates the fourth image by using a definition value calculation method to obtain a fourth definition value; first distance | (first definition value-third definition value)/(first definition value-second definition value) | × a;
comparing the fourth sharpness value with the third sharpness value;
if the fourth definition value is large, defining the fourth definition value as a third definition value, and entering a judging step;
if the fourth definition value is small, the control module controls the shooting module to move the second distance in the fourth direction and shoot to obtain a fifth image, and then the control module calculates the fifth image by using a definition value calculation method to obtain a fifth definition value; defining the fifth definition value as the third definition value, and entering a judging step;
the fourth direction is vertical to the platform where the shooting object is located;
the second pitch is greater than the first pitch.
15. A fast focus adjustment method, characterized in that the method comprises the steps of:
the method comprises the following steps that a control module controls a shooting module to respectively shoot a shooting object at an optimal focusing position and a preset position to obtain a first image and a second image, and then the control module respectively calculates the first image and the second image by using a definition value calculation method to obtain a corresponding first definition value and a corresponding second definition value; the control module controls the shooting module to return to the optimal focusing position; the optimal focusing position and the preset position are both positioned on a Z axis vertical to the plane of the shooting object, and the distance between the optimal focusing position and the preset position is A;
a monitoring step, wherein the monitoring step comprises a photographing step, a judging step and a focusing step;
in the step of photographing, the control module controls the photographing module to photograph to obtain a third image, and then the control module calculates the third image by using a definition value calculation method to obtain a third definition value;
in the judging step, if the difference value between the first definition value and the third definition value does not exceed the threshold range, focusing is finished; if the difference value between the first definition value and the third definition value exceeds the threshold value range, entering a focusing step;
in the focusing step, the control module controls the shooting module to move the first distance to the fifth direction and shoot to obtain a fourth image, and then the control module calculates the fourth image by using a definition value calculation method to obtain a fourth definition value; first distance | (first definition value-third definition value)/(first definition value-second definition value) | × a;
comparing the fourth sharpness value with the third sharpness value;
if the fourth definition value is large, defining the fourth definition value as a third definition value, and entering a judging step;
if the fourth definition value is small, the control module controls the shooting module to move the second distance in the sixth direction and shoot to obtain a fifth image, and then the control module calculates the fifth image by using a definition value calculation method to obtain a fifth definition value; defining the fifth definition value as a third definition value, defining the sixth direction as a fifth direction, and entering a judging step;
the fifth direction and the sixth direction are both vertical to a platform where a shooting object is located, and the fifth direction is opposite to the sixth direction;
the second pitch is greater than the first pitch.
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