CN113916779B - Microscopic image acquisition control method, microscopic image acquisition control system and computer-readable storage medium - Google Patents
Microscopic image acquisition control method, microscopic image acquisition control system and computer-readable storage medium Download PDFInfo
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Abstract
The invention discloses a microscopic image acquisition control method for rapid and accurate focusing, which comprises the following steps: s1, acquiring plane coordinate information of a reference point and at least two preselected points from a currently acquired slide sample image; s2, controlling the objective lens to focus the reference point on the slide glass sample in a full range along the Z-axis direction, and obtaining initial coordinate information after focusing; s3, controlling the objective lens to respectively adjust and focus the reference point and the at least two preselected points on the slide glass sample along the Z-axis direction according to the plane coordinate information and the Z-axis information in the initial coordinate information to obtain at least three pieces of adjustment coordinate information; s4, constructing a fitting plane of the slide sample according to the at least three pieces of adjustment coordinate information, and then carrying out image scanning on the slide sample on the fitting plane, so that the invention can carry out rapid and accurate scanning on a sample image.
Description
Technical Field
The invention relates to the field of microscopic photographing, in particular to a microscopic image acquisition control method, a control system and a computer readable storage medium for rapid and accurate focusing.
Background
The pathological section is that a part of diseased tissue or viscera is processed by various chemicals and burying methods, so that the diseased tissue or viscera is fixed and hardened, the diseased tissue or viscera is cut into slices on a slicing machine, the slices are adhered on a slide, and the slices are dyed with various colors for examination under a microscope to observe pathological changes, so that pathological diagnosis is made, help is provided for clinical diagnosis and treatment, along with the progress of science and technology, people combine a microscope with a scanner, and microscopic images of the pathological section are displayed through a display screen, so that the pathological section is convenient for doctors to check.
In order to improve the detection efficiency, most of the mechanisms at present successively adopt an automatic image acquisition method for microscopic photographing of pathological sections, wherein the automatic image acquisition method combines optical, mechanical and electrical technologies with a cytological diagnosis technology, and whether cells are abnormal or cancerous is judged by measuring a glass slide loaded with the pathological sections so as to make pathological diagnosis. Although the existing automatic image acquisition method can acquire images, improves the detection accuracy and the detection efficiency, in the actual microscopic shooting process, the focusing process is too long and the situation of unclear focusing can also occur frequently, so that the rapid and accurate focusing of sample images is still a key point of continuous improvement, the success or failure of sample image scanning is directly influenced by the focusing quality, and the detection result is greatly disturbed.
In view of this, it is necessary to provide a fast and accurate focusing microscopic image acquisition control method to solve or at least alleviate the above-mentioned drawbacks of slow focusing process and unclear focusing.
Disclosure of Invention
Aiming at the technical problems, the main purpose of the invention is to provide a microscopic image acquisition control method for rapid and accurate focusing.
In order to achieve the above object, the present invention provides a method for controlling the acquisition of microscopic images with rapid and accurate focusing, comprising the steps of:
s1, acquiring plane coordinate information of a reference point and at least two preselected points from a currently acquired slide sample image, wherein the plane coordinate information is information of X-axis and Y-axis corresponding to the reference point and the at least two preselected points; wherein the reference point and the at least two preselected points are located on at least two different straight lines when at the same height, and the slide sample is placed between a light source and an objective lens of the microscopy device;
s2, controlling the objective lens to perform full-range focusing on the reference point on the slide glass sample along the Z-axis direction to obtain initial coordinate information after focusing, wherein the initial coordinate information is information of an X-axis, a Y-axis and a Z-axis of a focusing position corresponding to the reference point after full-range focusing;
s3, controlling the objective lens to adjust and focus the reference point and the at least two preselected points on the slide sample along the Z-axis direction according to the plane coordinate information and the Z-axis information in the initial coordinate information to obtain at least three pieces of adjustment coordinate information, wherein the adjustment coordinate information is information of an X axis, a Y axis and a Z axis corresponding to the reference point and the at least two preselected points after adjustment and focusing;
s4, constructing a fitting plane of the slide sample according to the at least three pieces of adjustment coordinate information, and then carrying out image scanning on the slide sample on the fitting plane.
Further, in the step S1, the step of acquiring the reference point and the pre-selected point includes:
s11, identifying a plurality of convex hull areas in the slide sample image;
s12, constructing a plurality of concentric circles according to a plurality of convex hull areas;
s13, selecting a reference circle from a plurality of concentric circles according to the number of the convex hull areas penetrated by each concentric circle and the dispersion degree of the convex hull areas on each concentric circle;
s14, selecting at least three scattered convex hull areas on a plurality of convex hull areas penetrated by the reference circle or on a plurality of convex hulls closest to the reference circle as point selection areas, then respectively selecting a point from a sample image in each point selection area as a base point, and taking the base point as the reference point or the preselected point.
Further, the step of controlling the objective lens in the step S2 to perform full-range focusing on the reference point on the slide sample along the Z-axis direction includes:
s21, acquiring a preset distance range between the reference point on the slide sample and the objective lens in the Z-axis direction;
s22, controlling the relative distance between the reference point on the slide glass sample and the objective lens to change within the preset distance range, and judging the initial coordinate information corresponding to the reference point after full-range focusing according to the real-time definition value in the changing process.
Further, the step S21 further includes: acquiring a preset definition value of the slide glass sample;
in the step S22, the determining, according to the real-time sharpness value in the changing process, the initial coordinate information corresponding to the reference point after the full-range focusing includes:
when the corresponding real-time definition value in the changing process is smaller than the preset definition value, the relative distance between the reference point and the objective lens is determined to not reach the full-range focusing standard;
and when the real-time definition corresponding to the change process is not smaller than the preset definition value, determining that the relative distance between the reference point and the objective lens reaches the standard of full-range focusing, and acquiring the initial coordinate information corresponding to the full-range focusing.
Further, the adjusting focus comprises the steps of:
s31, controlling the objective lens to move to a designated position according to the Z-axis information of the initial coordinate information and the plane coordinate information;
s32, controlling the objective lens to move downwards along the Z-axis direction by a preset distance, judging the focusing movement direction of the objective lens according to the definition change result when the objective lens moves downwards, and obtaining the adjustment coordinate information through the definition change result generated in the focusing movement process of the objective lens.
Further, the determining the focus moving direction of the objective lens according to the change result of the definition in the downward movement includes:
when the change result of the definition is that the definition is reduced during the downward movement, the focusing movement direction of the objective lens is that the objective lens is moved upwards until the definition is not increased any more;
when the change of the definition is raised as the downward movement, the focus movement direction of the objective lens is moved downward until the definition is no longer raised.
Further, in the step S4, the constructing a fitting plane of the slide sample according to the at least three adjustment coordinate information includes: and performing secondary polynomial surface fitting on the at least three pieces of adjustment coordinate information to obtain the fitting plane.
Further, in the process of the full-range focusing and the focusing adjustment, the coordinate switching is performed by controlling the movement of the objective lens in the X-axis, Y-axis and Z-axis directions;
the minimum objective lens moving unit for full-range focusing is larger than the minimum objective lens moving unit for adjusting focusing, wherein the minimum objective lens moving unit is the minimum distance for moving the objective lens along the Z-axis direction in the focusing process.
The invention also provides a control system of a microscopic device, comprising a memory, a processor, a computer program stored in the memory and running on the processor, and a controller receiving instructions from the processor, wherein the processor executes the computer program to implement the steps of the image acquisition method according to any one of the above.
The present invention also provides a computer-readable storage medium storing a computer program which, when executed by a processor, implements the steps of the image acquisition method as described in any one of the above
Compared with the prior art, the invention has the following advantages:
the invention provides a microscopic image acquisition control method for rapid and accurate focusing, which can rapidly and accurately focus a sample pattern. By acquiring one reference point and carrying out full-range focusing on the position of the reference point, initial coordinate information can be obtained, and a base and a reference are provided for follow-up accurate adjustment focusing; by acquiring at least two pre-selected points, under the condition of matching the reference points and combining the initial coordinate information, the reference points and the at least two pre-selected points can be adjusted and focused, so that accurate focuses at the reference points and the pre-selected points are obtained, and at least three pieces of adjusted coordinate information are obtained; by acquiring the adjusted coordinate information and performing a plane fit on the slide sample with the adjusted coordinate information, an accurate image scan plane can be obtained.
In addition, in the process of acquiring the reference point and the preselected point, the convex hull area and the concentric circle are introduced, and the reference circle and the selected point area are identified, so that the finally fitted plane can reflect the plane of the slide sample image to the greatest extent, and a clearer image can be obtained through subsequent scanning; in the focusing adjustment process, the focusing movement direction of the objective lens is judged according to the definition change result when the objective lens moves downwards, and the adjustment coordinate information can be quickly and accurately obtained through the definition change result generated in the focusing movement process of the objective lens; by adopting the quadratic polynomial surface fitting, a more accurate fitting plane can be obtained by means of the adjustment coordinate information.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings that are required in the embodiments or the description of the prior art will be briefly described, and it is obvious that the drawings in the following description are only some embodiments of the present invention, and other drawings may be obtained according to the structures shown in these drawings without inventive effort for a person skilled in the art.
FIG. 1 is a flow chart of a microscopic image acquisition control method according to an embodiment of the present invention;
FIG. 2 is a flow chart of step S1 according to an embodiment of the present invention;
FIG. 3 is a flow chart of step S2 according to an embodiment of the present invention;
FIG. 4 is a flow chart of step S3 according to an embodiment of the present invention;
FIG. 5 is a schematic illustration of the selection of reference points and preselected points in one embodiment of the invention.
Reference numerals illustrate: a slide sample image 1, a convex hull area 2, concentric circles 3, a reference circle 4, a point selection area 5 and a base point 6.
The achievement of the object, functional features and advantages of the present invention will be further described with reference to the drawings in connection with the embodiments.
Detailed Description
The following description of the embodiments of the present invention will be made clearly and fully with reference to the accompanying drawings, in which it is evident that the embodiments described are only some, but not all embodiments of the invention. All other embodiments, based on the embodiments of the invention, which are apparent to those of ordinary skill in the art without inventive faculty, are intended to be within the scope of the invention.
It should be noted that all directional indicators (such as upper and lower … …) in the embodiments of the present invention are merely used to explain the relative positional relationship, movement conditions, etc. between the components in a specific posture (as shown in the drawings), and if the specific posture is changed, the directional indicator is changed accordingly.
Furthermore, descriptions such as those referred to as "first," "second," and the like, are provided for descriptive purposes only and are not to be construed as indicating or implying a relative importance or implying an order of magnitude of the indicated technical features in the present disclosure. Thus, a feature defining "a first" or "a second" may explicitly or implicitly include at least one such feature.
Moreover, the technical solutions of the embodiments of the present invention may be combined with each other, but it is necessary to be based on the fact that those skilled in the art can implement the embodiments, and when the technical solutions are contradictory or cannot be implemented, it should be considered that the combination of the technical solutions does not exist, and is not within the scope of protection claimed by the present invention.
Referring to fig. 1 to 5, in order to improve the scanning efficiency and accuracy, the invention provides a fast and accurate focusing microscopic image acquisition control method, which comprises the following steps:
s1, acquiring plane coordinate information of a reference point and at least two preselected points from a currently acquired slide sample image 1, wherein the plane coordinate information is information of an X axis and a Y axis corresponding to the reference point and the at least two preselected points; wherein the reference point and the at least two preselected points are located on at least two different straight lines when at the same height, and the slide sample is placed between a light source and an objective lens of the microscopy apparatus. It should be noted that the slide sample image refers to an image of a sample on the slide that is collected, and the number of the pre-selected points must be not less than two, however, in order to better reflect the entire plane of the slide sample image 1, it may also be not less than 5, not less than 6, not less than 10, etc., and specifically may be set according to practical situations.
Wherein, in order to cover the slide sample image 1 with the maximum range of the selected point position and embody the whole plane where the slide sample image 1 is located to the maximum extent, the step of obtaining the reference point and the pre-selected point may include:
s11, identifying a plurality of convex hull areas 2 in the slide sample image 1;
s12, constructing a plurality of concentric circles 3 according to a plurality of convex hull areas 2;
s13, selecting one reference circle 4 from a plurality of concentric circles 3 according to the number of the convex hull areas 2 penetrated by each concentric circle 3 and the dispersion degree of the convex hull areas 2 on each concentric circle;
s14, selecting at least three scattered convex hull areas 2 on a plurality of convex hull areas 2 penetrated by the reference circle 4 or on a plurality of convex hulls closest to the reference circle 4 as point selection areas 5, then selecting a point from a sample image in each point selection area 5 as a base point 6, and taking the base point 6 as the reference point or the preselected point.
It should be noted that, the convex hull region 2 refers to a region irradiated by a plane convex hull in the slide sample image 1, the plane convex hull is a protrusion on an outer contour of the slide sample image 1, and the concentric circle 3 is constructed by taking the convex hull region 2 as a reference, so that the sample image can be covered in a maximum range, and the whole plane of the carrier sample is reflected to the greatest extent when the subsequent plane is fitted. When the concentric circles 3 are constructed, it is preferable that a middle point of the slide sample image 1 is used as a center, and when the exact middle point cannot be determined, a point may be selected as a center in a middle area of the slide sample image 1 because the slide sample image 1 is irregular.
However, since the slide sample image 1 is generally irregular and the positions of the projections formed on the plane are also disordered, although the concentric circle 3 at the outermost periphery has the widest coverage, most of the area through which it passes may be an image on a blank slide, and therefore, the concentric circle 3 at the outermost periphery cannot be directly used as the reference circle 4, but should be selected in a targeted manner according to the step S13. Among the concentric circles 3, when the number of the convex hull regions 2 penetrating one of the concentric circles 3 is the largest and the degree of dispersion of each convex hull region 2 on the concentric circle is high, it can be used as the reference circle 4. The degree of dispersion is a concept of reference such as: when the convex hull regions 2 penetrated by a certain concentric circle 3 are the most, the dispersion degree of each convex hull region 2 on the concentric circle needs to be identified, and if each convex hull region 2 on the concentric circle is found to be extruded in a stack, the judgment needs to be carried out again after the concentric circle 3 is removed. Moreover, when the number of the convex hull regions 2 penetrated by several concentric circles 3 is identical, the concentric circle 3 with the highest dispersion degree of the convex hull regions 2 needs to be selected as the reference circle 4. It should be noted that, in an extreme case, when the number of the convex hull regions 2 penetrated by the two concentric circles 3 is identical, and the dispersion degree of each of the convex hull regions 2 on the two concentric circles is also close, the concentric circle on the outer side may be selected as the reference circle 4. After the reference circle 4 is determined, the reference circle 4 may be used as a reference for selecting points, the reference circle 4 may be used for selecting points in the convex hull region 2 penetrated by the reference circle 4, in order to improve the dispersion degree of the selected points, the reference circle 4 may be used for selecting points nearby, or the reference circle 4 may be used for selecting points in the nearby convex hull region 2, and several modes may be combined with each other. It should be clear that, at a specific point of selection, both the reference point and the preselected point selected need to fall on the sample image.
The reference point mainly has the function of providing a focused plane locus for the whole range, namely, coarse focusing is carried out at the position of the reference point, and the reference point can be used as one of the plane loci for adjusting focusing continuously in the later focusing adjustment process; the preselected points are primarily planar sites that provide focus for adjusting focus. In addition, since the reference point and the preselected point are mainly focused plane sites, only plane coordinate information is needed to be considered when the reference point and the preselected point are selected; in addition, since the fitting is required to be performed on the plane later, when the reference point and the preselected points are selected, the reference point and the at least two preselected points are located on at least two different straight lines when located at the same height, specifically, a virtual height can be set, and plane coordinate information of the reference point and the at least two preselected points can be acquired according to the same height standard.
In addition, it should be appreciated by those skilled in the art that in a specific application scenario, the placement of the slide sample between the light source and the objective lens of the microscope device is a conventional option, and in the present invention, the stage carrying the slide sample may be fixed during the subsequent focusing process, and the coordinate switching may be achieved by controlling the movement of the objective lens.
S2, controlling the objective lens to perform full-range focusing on the reference point on the slide glass sample along the Z-axis direction to obtain initial coordinate information after focusing, wherein the initial coordinate information is information of an X-axis, a Y-axis and a Z-axis of a focusing position corresponding to the reference point after full-range focusing.
It should be noted that in the process of full-range focusing, the objective lens needs to be adjusted to be above the reference point, and the distance between the objective lens and the glass slide sample is controlled along the Z-axis direction, so that Z-axis information after full-range focusing is obtained, and the initial coordinate information can be obtained by combining the X-axis information and the Y-axis information, and the initial coordinate information is used as a reference in the subsequent focusing process. In addition, since the stage may be fixedly disposed, the Z-axis information may be used primarily to refer to the relative distance between the objective lens and the slide sample along the Z-axis direction.
Specifically, the step of controlling the objective lens to perform the full-range focusing on the reference point on the slide sample along the Z-axis direction in the step S2 may include:
s21, acquiring a preset distance range between the reference point on the slide sample and the objective lens in the Z-axis direction;
s22, controlling the relative distance between the reference point on the slide glass sample and the objective lens to change within the preset distance range, and judging the initial coordinate information corresponding to the reference point after full-range focusing according to the real-time definition value in the changing process.
As a further explanation of the step S21, the step S21 further includes: acquiring a preset definition value of the slide glass sample;
as a refinement of the step S22, in the step S22, the determining, according to the real-time sharpness value in the changing process, the initial coordinate information corresponding to the reference point after focusing in the full range includes:
when the corresponding real-time definition value in the changing process is smaller than the preset definition value, the relative distance between the reference point and the objective lens is determined to not reach the full-range focusing standard;
and when the real-time definition corresponding to the change process is not smaller than the preset definition value, determining that the relative distance between the reference point and the objective lens reaches the standard of full-range focusing, and acquiring the initial coordinate information corresponding to the full-range focusing.
In addition, to facilitate a further understanding of the criteria for full-range focusing, the step S22 may further include:
and controlling the relative distance between the reference point and the objective lens to be sequentially changed from small to large, and recognizing that the relative distance between the reference point and the objective lens reaches the full-range focusing standard when a first real-time definition value larger than a preset definition value is generated in the process of sequentially changing the relative distance from small to large.
S3, controlling the objective lens to adjust and focus the reference point and the at least two preselected points on the slide sample along the Z-axis direction according to the plane coordinate information and the Z-axis information in the initial coordinate information to obtain at least three pieces of adjustment coordinate information, wherein the adjustment coordinate information is information of an X axis, a Y axis and a Z axis corresponding to the reference point and the at least two preselected points after adjustment and focusing.
It should be noted that, after the initial coordinate information is obtained, the Z-axis information of the initial coordinate information and the plane coordinate information of the reference point and the preselected point may be used as references, and by controlling the movement of the objective lens, the reference point and the preselected point on the slide sample may be accurately focused for fitting the plane to be scanned.
In order to be able to obtain the adjustment coordinate information quickly and accurately, the adjustment focus may comprise the steps of:
s31, controlling the objective lens to move to a designated position according to the Z-axis information of the initial coordinate information and the plane coordinate information; the designated position may include a position pointed by the initial coordinate information and a position shifted from the position of the initial coordinate information to the position after the selected point;
s32, controlling the objective lens to move downwards along the Z-axis direction by a preset distance, judging the focusing movement direction of the objective lens according to the definition change result when the objective lens moves downwards, and obtaining the adjustment coordinate information through the definition change result generated in the focusing movement process of the objective lens, namely, obtaining the coordinate information corresponding to the definition reaching the preset condition as the adjustment coordinate information.
Specifically, the determining the focus moving direction of the objective lens according to the change result of the definition in the downward movement may include:
when the change result of the definition is that the definition is reduced during the downward movement, the focusing movement direction of the objective lens is that the objective lens is moved upwards until the definition is not increased any more, and the corresponding coordinate information is the adjustment coordinate information at the moment;
and when the change result of the definition is that the definition is increased during the downward movement, the focusing movement direction of the objective lens is that the objective lens is moved downwards until the definition is not increased any more, and the corresponding coordinate information is the adjustment coordinate information at the moment.
S4, constructing a fitting plane of the slide sample according to the at least three pieces of adjustment coordinate information, and then carrying out image scanning on the slide sample on the fitting plane. It should be appreciated that the fitting plane may be constructed by a fitting method conventional in the art, and preferably, the constructing the fitting plane of the slide sample according to the at least three adjustment coordinate information may also include: and performing secondary polynomial surface fitting on the at least three pieces of adjustment coordinate information to obtain the fitting plane.
The quadratic polynomial surface formula adopted by the quadratic polynomial surface fitting is as follows:
f（x,y）=a
20
x
2
+a
02
y
2
+a
11
xy+a
10
x+a
01
y+a
00
wherein the above formulae are commona 20 、a 02 、a 11 、a 10 、a 01 、a 00 And 6 coefficients. In addition, in the case of the optical fiber,f（x,y）may be expressed as a number corresponding to the Z axis,xmay be expressed as a number corresponding to the X-axis,ymay be expressed as a value corresponding to the Y-axis.
As a description of the above embodiments, in the process of the full-range focusing and the adjustment focusing, the switching of coordinates may be performed by controlling the movement of the objective lens in the X-axis, Y-axis, and Z-axis directions.
In addition, the minimum objective lens moving unit for full-range focusing is larger than the minimum objective lens moving unit for adjusting focusing, wherein the minimum objective lens moving unit is the minimum distance for moving the objective lens along the Z-axis direction in the focusing process.
The invention also provides a control system of a microscopic device, which comprises a memory, a processor, a computer program stored in the memory and capable of running on the processor, and a controller for receiving instructions of the processor, wherein the steps of the image acquisition method according to any one of the embodiments are realized when the processor executes the computer program.
The present invention also provides a computer readable storage medium storing a computer program which, when executed by a processor, implements the steps of the image acquisition method according to any one of the embodiments described above.
In the above technical solution of the present invention, the above is only a preferred embodiment of the present invention, and therefore, the patent scope of the present invention is not limited thereto, and all the equivalent structural changes made by the description of the present invention and the content of the accompanying drawings or the direct/indirect application in other related technical fields are included in the patent protection scope of the present invention.
Claims (9)
1. A microscopic image acquisition control method for rapid and accurate focusing is characterized by comprising the following steps:
s1, acquiring plane coordinate information of a reference point and at least two preselected points from a currently acquired slide sample image, wherein the plane coordinate information is information of X-axis and Y-axis corresponding to the reference point and the at least two preselected points; wherein the reference point and the at least two preselected points are located on at least two different straight lines when at the same height, and the slide sample is placed between a light source and an objective lens of the microscopy device;
the acquisition of the slide sample image irregularities, the reference points and the pre-selected points comprises the sub-steps of:
s11, identifying a plurality of convex hull areas in the slide sample image;
the convex hull region is a region irradiated by a plane convex hull in the slide sample image, and the plane convex hull is a bulge on the outer contour of the slide sample image;
s12, constructing a plurality of concentric circles according to a plurality of convex hull areas;
s13, selecting a reference circle from a plurality of concentric circles according to the number of the convex hull areas penetrated by each concentric circle and the dispersion degree of the convex hull areas on each concentric circle;
s14, selecting at least three scattered convex hull areas as point selection areas on a plurality of convex hull areas penetrated by the reference circle or on a plurality of convex hull areas closest to the reference circle, then selecting a point from a sample image in each point selection area as a base point, and taking the base point as the reference point or the preselected point;
s2, controlling the objective lens to perform full-range focusing on the reference point on the slide glass sample along the Z-axis direction to obtain initial coordinate information after focusing, wherein the initial coordinate information is information of an X-axis, a Y-axis and a Z-axis of a focusing position corresponding to the reference point after full-range focusing;
s3, controlling the objective lens to adjust and focus the reference point and the at least two preselected points on the slide sample along the Z-axis direction according to the plane coordinate information and the Z-axis information in the initial coordinate information to obtain at least three pieces of adjustment coordinate information, wherein the adjustment coordinate information is information of an X axis, a Y axis and a Z axis corresponding to the reference point and the at least two preselected points after adjustment and focusing;
s4, constructing a fitting plane of the slide sample according to the at least three pieces of adjustment coordinate information, and then carrying out image scanning on the slide sample on the fitting plane.
2. The microscopic image acquisition control method according to claim 1, wherein the step of controlling the objective lens to perform full-range focusing on the reference point on the slide sample along the Z-axis direction in the step S2 includes:
s21, acquiring a preset distance range between the reference point on the slide sample and the objective lens in the Z-axis direction;
s22, controlling the relative distance between the reference point on the slide glass sample and the objective lens to change within the preset distance range, and judging the initial coordinate information corresponding to the reference point after full-range focusing according to the real-time definition value in the changing process.
3. The microscopic image acquisition control method according to claim 2, wherein the step S21 further includes: acquiring a preset definition value of the slide glass sample;
in the step S22, the determining, according to the real-time sharpness value in the changing process, the initial coordinate information corresponding to the reference point after the full-range focusing includes:
when the corresponding real-time definition value in the changing process is smaller than the preset definition value, the relative distance between the reference point and the objective lens is determined to not reach the full-range focusing standard;
and when the real-time definition corresponding to the change process is not smaller than the preset definition value, determining that the relative distance between the reference point and the objective lens reaches the standard of full-range focusing, and acquiring the initial coordinate information corresponding to the full-range focusing.
4. The microscopic image acquisition control method according to claim 1, wherein the adjusting focus includes the steps of:
s31, controlling the objective lens to move to a designated position according to the Z-axis information of the initial coordinate information and the plane coordinate information;
s32, controlling the objective lens to move downwards along the Z-axis direction by a preset distance, judging the focusing movement direction of the objective lens according to the definition change result when the objective lens moves downwards, and obtaining the adjustment coordinate information through the definition change result generated in the focusing movement process of the objective lens.
5. The microscopic image acquisition control method according to claim 4, wherein the judging a focus moving direction of the objective lens based on a change result of the sharpness when moving downward includes:
when the change result of the definition is that the definition is reduced during the downward movement, the focusing movement direction of the objective lens is that the objective lens is moved upwards until the definition is not increased any more;
when the change of the definition is raised as the downward movement, the focus movement direction of the objective lens is moved downward until the definition is no longer raised.
6. The microscopic image acquisition control method according to claim 1, wherein in the step S4, the constructing the fitting plane of the slide specimen according to the at least three pieces of adjustment coordinate information includes: and performing secondary polynomial surface fitting on the at least three pieces of adjustment coordinate information to obtain the fitting plane.
7. The microscopic image acquisition control method according to any one of claims 1-6, wherein the switching of coordinates is performed by controlling the movement of the objective lens in the X-axis, Y-axis, Z-axis directions during the full-range focusing and the adjustment focusing;
the minimum objective lens moving unit for full-range focusing is larger than the minimum objective lens moving unit for adjusting focusing, wherein the minimum objective lens moving unit is the minimum distance for moving the objective lens along the Z-axis direction in the focusing process.
8. A control system for a microscopy apparatus comprising a memory, a processor, a computer program stored in the memory and executable on the processor, and a controller receiving instructions from the processor, wherein execution of the computer program by the processor effects the steps of the microscopy image acquisition control method of any one of claims 1 to 7.
9. A computer-readable storage medium storing a computer program, wherein the computer program when executed by a processor implements the steps of the microscopic image acquisition control method according to any one of claims 1 to 7.
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