阅读说明：本技术 一种显微镜对焦方法、装置及显微镜设备 (Microscope focusing method and device and microscope equipment ) 是由 孙宝亮 郭灵犀 张鹏宇 张志龙 郑宇� 潘红九 马鸣 李萌萌 王保录 纪祖赑 赵 于 2019-10-29 设计创作，主要内容包括：一种显微镜对焦方法、装置及显微镜设备,包括：控制载物装置携带待测物体在预先确定的图像清晰经验范围内沿y轴方向移动所述待测物体,获取每个焦平面上所述待测物体的图像；所述y轴为垂直于显微镜焦平面的方向；根据所述待测物体的图像确定所要从所述待测物体的图像中截取的所述待测物体的经验区域图像；根据所述待测物体的经验区域图像计算反差值；根据反差值最大的位置将显微镜焦点置于该位置焦点上。采用本申请中的方案,只需要很少的“拉风箱”操作即可完成对焦,极大提高了对焦效率。(A microscope focusing method, a device and a microscope device comprise: controlling a carrying device to carry an object to be detected to move the object to be detected along the y-axis direction within a predetermined image definition experience range, and acquiring an image of the object to be detected on each focal plane; the y-axis is perpendicular to the focal plane of the microscope; determining an experience area image of the object to be detected, which is intercepted from the image of the object to be detected, according to the image of the object to be detected; calculating a contrast value according to the experience area image of the object to be detected; the microscope focus is placed at the position of the maximum contrast value. By adopting the scheme in the application, focusing can be completed only by few operations of 'air box pulling', and the focusing efficiency is greatly improved.)

1. A method of focusing a microscope, comprising:

controlling a carrying device to carry an object to be detected to move the object to be detected along the y-axis direction within a predetermined image definition experience range, and acquiring an image of the object to be detected on each focal plane; the y-axis is perpendicular to the focal plane of the microscope;

determining an experience area image of the object to be detected, which is intercepted from the image of the object to be detected, according to the image of the object to be detected;

calculating a contrast value according to the experience area image of the object to be detected;

the microscope focus is placed at the position of the maximum contrast value.

2. The method of claim 1, further comprising, before the controlling the object carrying device to move the object along the y-axis within a predetermined image sharpness experience range, the method further comprising:

and controlling the object carrying device to move in the y-axis direction until an image of the object to be detected appears in the visual field of an eyepiece of the microscope.

3. The method of claim 1, wherein the object to be measured is a cylindrical object, the side surface of the object to be measured is a curved surface, and the acquiring the image of the object to be measured on each focal plane comprises: and acquiring an image of the side surface of the object to be detected on each focal plane.

4. The method according to claim 1 or 3, wherein the microscope is a fluorescence microscope, the object to be measured is a medical probe, a stained cell is adsorbed on the medical probe, and the stained cell is stained with a fluorescent substance.

5. The method of claim 4, wherein the microscope illuminates the medical probe with a background light source.

6. The method according to claim 3, wherein the determining an image of the empirical region of the object to be captured from the image of the object to be captured comprises:

determining the edge of the object to be detected in the image of the object to be detected;

calculating the distance e from the edge of the experience area of the object to be measured to the edge of the object to be measured closest to the edge of the experience area;

obtaining the number of pixels from the edge of the experience area of the object to be measured to the edge of the object to be measured closest to the edge of the experience area according to the relation between the camera imaging pixel and the actual length of the object to be measured obtained through pre-measurement and the distance e;

and determining the empirical region image of the object to be detected, which is to be intercepted from the image of the object to be detected, according to the number of pixels from the edge of the empirical region of the object to be detected to the edge of the object to be detected, which is closest to the edge of the empirical region.

7. The method according to claim 6, wherein the distance e from the edge of the empirical region of the object to the edge of the object closest to the edge of the empirical region is calculated according to the following formula:

wherein d is the diameter of the cylindrical object to be measured; h is the depth of field.

8. The method according to claim 6, wherein the empirical region of the object under test corresponds to a sector angle α of the cross section of the object under test, where α is:

α＝2arccos(d-h/d)；

wherein d is the diameter of the cylindrical object to be measured; h is the depth of field.

9. The method of claim 1, wherein the determination of the clear empirical range of the image is performed as follows:

comparing the image of the object with the same type as the object to be detected with a predetermined experience area of the object with the same type as the object to be detected;

and determining the clear experience range of the image of the movement of the focusing axis when the microscope shoots the object of the type according to the comparison result.

10. A microscope focusing device, comprising:

the first control module is used for controlling the object carrying device to carry the object to be detected to move the object to be detected along the y-axis direction within a predetermined image definition experience range;

the image acquisition module is used for acquiring an image of the object to be detected on each focal plane; the y-axis is perpendicular to the focal plane of the microscope;

the first calculation module is used for determining an experience area image of the object to be detected, which is intercepted from the image of the object to be detected, according to the image of the object to be detected;

the second calculation module is used for calculating a contrast value according to the experience area image of the object to be detected;

and the second control module is used for placing the microscope focus on the focus at the position according to the position with the maximum contrast value.

11. The apparatus of claim 10, further comprising:

and the third control module is used for controlling the object carrying device to carry the object to be detected to move along the y-axis direction within a predetermined image definition experience range before the object to be detected, and controlling the object carrying device to carry the object to be detected to move in the y-axis direction until the image of the object to be detected appears in the visual field of an eyepiece of the microscope.

12. The apparatus according to claim 10, wherein the object to be measured is a cylindrical object, the side surface of the object to be measured is a curved surface, and the image obtaining module is configured to obtain an image of the side surface of the object to be measured at each focal plane.

13. The apparatus of claim 11, wherein the first computing module comprises:

the first determining unit is used for determining the edge of the object to be detected in the image of the object to be detected;

a first calculation unit, configured to calculate a distance e from an edge of an experience area of the object to be measured to an edge of the object to be measured closest to the edge of the experience area;

the second calculation unit is used for obtaining the number of pixels from the edge of the experience area of the object to be measured to the edge of the object to be measured closest to the edge of the experience area according to the relation between the camera imaging pixel and the actual length of the object to be measured obtained through measurement in advance and the distance e;

and the second determining unit is used for determining the experience area image of the object to be detected, which is to be intercepted from the image of the object to be detected, according to the number of pixels from the edge of the experience area of the object to be detected to the edge of the object to be detected, which is closest to the edge of the experience area.

14. A microscope apparatus, comprising: an image acquisition device and a carrying device of a microscope, and a microscope focusing device according to any one of claims 10 to 13; wherein the content of the first and second substances,

the image acquisition device of the microscope is used for shooting the image of the object to be measured under the control of the microscope focusing device;

the object carrying device is used for carrying the object to be detected to move under the control of the microscope focusing device.

Technical Field

The present application relates to a fluorescence microscope focusing technology, and in particular, to a microscope focusing method, device and microscope equipment.

Background

The fluorescence microscope equipment is mostly used for clinical pathological section scanning, focusing is needed in the pathological section scanning process, the clearest point in an image is found, so that a clear and complete pathological section scanning image is obtained, and focusing is usually realized by adopting an inverse differential focusing technology at present.

The focusing process of the contrast focusing technology is as follows:

the method comprises the steps of driving a lens, changing focus points along an axis pointing to a shot object, acquiring an image on each focus point, similarly to point-by-point scanning, digitizing the image acquired on each focus point (the digitized image is an integer matrix) and transmitting the image to an image processor, calculating the contrast amount (or called definition), screening out the focus point with the maximum contrast through comparison, driving the lens to place the focus point on the focus point with the maximum contrast to obtain a correct focus point, determining whether to focus according to the contrast value with the maximum contrast amount, and finishing focusing.

The above process is reflected on the screen, and is a 'bellows' process from blurring to blurring and then blurring to final blurring.

However, as can be seen from the above description of the focusing process, it is necessary to scan point by point, digitize each focus, calculate the contrast amount, and finally screen the focus with the highest contrast, and the entire focusing process takes a long time.

Disclosure of Invention

The embodiment of the application provides a microscope focusing method, a microscope focusing device and microscope equipment, and aims to solve the technical problems.

According to a first aspect of embodiments of the present application, there is provided a microscope focusing method, including the steps of:

controlling a carrying device to carry an object to be detected to move the object to be detected along the y-axis direction within a predetermined image definition experience range, and acquiring an image of the object to be detected on each focal plane; the y-axis is perpendicular to the focal plane of the microscope;

determining an experience area image of the object to be detected, which is intercepted from the image of the object to be detected, according to the image of the object to be detected;

calculating a contrast value according to the experience area image of the object to be detected;

the microscope focus is placed at the position of the maximum contrast value.

According to a second aspect of embodiments of the present application, there is provided a microscope focusing device, comprising:

the first control module is used for controlling the object carrying device to carry the object to be detected to move the object to be detected along the y-axis direction within a predetermined image definition experience range;

the image acquisition module is used for acquiring an image of the object to be detected on each focal plane; the y-axis is perpendicular to the focal plane of the microscope;

the first calculation module is used for determining an experience area image of the object to be detected, which is intercepted from the image of the object to be detected, according to the image of the object to be detected;

the second calculation module is used for calculating a contrast value according to the experience area image of the object to be detected;

and the second control module is used for placing the microscope focus on the focus at the position according to the position with the maximum contrast value.

According to a third aspect of embodiments of the present application, there is provided a microscope apparatus characterized by comprising: the microscope comprises an image acquisition device, a carrying device and the microscope focusing device; wherein the content of the first and second substances,

the image acquisition device of the microscope is used for shooting the image of the object to be measured under the control of the microscope focusing device;

the object carrying device is used for carrying the object to be detected to move under the control of the microscope focusing device.

By adopting the microscope focusing method, the microscope focusing device and the microscope equipment, the clear experience range of the image is predetermined, the object to be detected is controlled to move along the focusing axis (y axis) direction according to the experience range, the experience area image of the object to be detected is obtained by intercepting after the image of the object to be detected is obtained, the contrast value is calculated according to the experience area image, the focus of the microscope is placed on the focus of the position according to the position with the maximum contrast value, so that focusing can be completed only by few operations of a drawing bellows, and the focusing efficiency is greatly improved.

Drawings

The accompanying drawings, which are included to provide a further understanding of the application and are incorporated in and constitute a part of this application, illustrate embodiment(s) of the application and together with the description serve to explain the application and not to limit the application. In the drawings:

FIG. 1 is a schematic flow chart illustrating an implementation of a microscope focusing method according to a first embodiment of the present application;

FIG. 2 is a schematic structural diagram of a microscope focusing device according to a second embodiment of the present application;

FIG. 3 is a schematic structural diagram of a microscope device according to a third embodiment of the present application;

FIG. 4 is an exploded view of a lens of a fluorescence microscope according to a fourth embodiment of the present application;

FIG. 5 is a schematic structural view of a fluorescence microscope in example four of the present application;

FIG. 6 is a schematic structural diagram of a loading device in the fourth embodiment of the present application;

FIG. 7 is a schematic structural diagram of an adjusting device in the fourth embodiment of the present application;

FIG. 8 is a schematic structural diagram of a clamping assembly in the fourth embodiment of the present application;

FIG. 9 is a schematic structural diagram of an optical device according to a fourth embodiment of the present application;

FIG. 10 shows an enlarged schematic view of a medical probe according to a fourth embodiment of the present application;

FIG. 11 is a schematic diagram showing the imaging of a sharp image experience zone in the fourth embodiment of the present application;

FIG. 12 is a schematic diagram showing the relationship between the cross section and the radial direction of the empirical region of the sharp image in the fourth embodiment of the present application;

1. a base; 2. a carrying device; 3. a drive device; 4. a light emitting device; 5. an optical device; 6. an image acquisition device;

21. an adjustment device; 22. a clamping assembly;

211. the objective table supports the guide rail frame; 212. an objective table reference seat; 213. a stage reference plate; 2141. a rotating electric machine; 2142. an X-direction linear motor; 2143. a Y-direction linear motor; 215. a rotating table; 216. a rotating table bracket; 217. a coupling; 218. an object stage;

221. clamping the substrate; 222. a first load bearing structure; 223. a second load bearing structure; 224. a first fixed structure; 225. a second fixed structure; 226. an object to be measured;

51. a disc-shaped bracket assembly; 52. a rotating shaft; 53. a lens barrel; 54. an element support.

Detailed Description

Aiming at the technical problems in the prior art, the embodiment of the application provides a microscope focusing method, a microscope focusing device and microscope equipment, which can realize the automatic focusing of a microscope, and provides an empirical zone calculation method aiming at the problem of single objective curved surface focusing.

The scheme in the embodiment of the application can be implemented by adopting various computer languages, such as object-oriented programming language C # and interpreted scripting language JavaScript.

In order to make the technical solutions and advantages of the embodiments of the present application more apparent, the following further detailed description of the exemplary embodiments of the present application with reference to the accompanying drawings makes it clear that the described embodiments are only a part of the embodiments of the present application, and are not exhaustive of all embodiments. It should be noted that the embodiments and features of the embodiments in the present application may be combined with each other without conflict.