Focal length value acquisition method, focal length value acquisition device and detection equipment

Abstract

The invention discloses a focal length value acquisition method, a focal length value acquisition device and detection equipment capable of calculating the focal length of a slide. The focal length value acquisition method comprises the following steps: acquiring a correction slope value and a correction intercept value of a detection device comprising a microscope; obtaining a height value of the slide by judging a scale when the probe is in contact with the carrying table and another scale when the probe is in contact with the slide according to the image about the carrying table; and calculating the calculated focal length of the slide according to the corrected slope value, the corrected intercept value and the height value.

Description

Technical Field

[0001] The present disclosure relates to a method for obtaining a focal length value, an apparatus for obtaining a focal length value, and a detection device capable of calculating the focal length of a glass slide, particularly in the field of microscope measurement. Background Art

[0002] In existing biological detection methods, particularly in circulating tumor cell (CTC) detection, steps such as cell separation, fluorescence staining, image acquisition, and result interpretation are usually automated processes that no longer require manual intervention. However, due to different manufacturing tolerances of different glass slides, when using different glass slides for detection, the tolerances will affect the shooting focus of the microscope, resulting in failed focusing of the microscope and thus causing the problem of blurred captured images.

[0003] Therefore, a method and an apparatus are needed to solve the above problems. Summary of the Invention

[0004] The present disclosure relates to a method for obtaining a focal length value, including: obtaining a focus correction slope value and a correction intercept value of a detection device including a microscope; obtaining a height value of a glass slide by judging a scale where a probe contacts a stage and another scale where the probe contacts the glass slide according to a plurality of images of the stage; and calculating a calculated focal length value of the glass slide according to the correction slope value, the correction intercept value, and the height value.

[0005] The present disclosure also relates to an apparatus for obtaining a focal length value, including a main body, a stage, a linear movement module, a probe, an optical scale module, an imaging module, and a processor. The main body includes a side surface. The stage is located on one side of the side surface and has a tabletop for placing a glass slide. The linear movement module is connected to the main body. The probe can move relative to the side surface closer to or farther from the stage through the linear movement module. The optical scale module includes a scale slide rail provided on the side surface and an optical scale reader / writer head moving along the scale slide rail. The optical scale reader / writer head is connected to the probe and is configured to read the scale of the scale slide rail. The imaging module is configured to capture images of the tabletop and output the images. The processor is configured to perform the following operations: controlling the linear movement module to move the probe along the scale slide rail towards the stage; controlling the imaging module to capture a plurality of images of the stage; calculating a height value of the glass slide by judging a scale where the probe contacts the stage and another scale where the probe contacts the glass slide according to the images of the stage; and calculating the calculated focal length of the glass slide according to the correction slope value of the detection device including the microscope, the correction intercept value of the detection device including the microscope, and the height value.

[0006] The present disclosure also relates to a detection device capable of calculating the focal length of a glass slide for detecting a biological sample located on the glass slide. The detection device includes the focal length value acquisition device and the microscope module as described above. The focal length value acquisition device is configured to obtain the calculated focal length value of the glass slide. The microscope module is configured to perform focusing and shooting according to the calculated focal length value. BRIEF DESCRIPTION OF THE DRAWINGS

[0007] Figure 1 It is a schematic diagram of the steps of a method for acquiring a focal length value according to an embodiment of the present disclosure.

[0008] Figure 2 It is a schematic structural diagram of a focal length value acquisition device according to an embodiment of the present disclosure.

[0009] Figure 3 It is a functional architecture diagram of a focal length value acquisition device according to an embodiment of the present disclosure.

[0010] Figure 4A It is a schematic diagram of a glass slide according to an embodiment of the present disclosure.

[0011] Figure 4B It is a schematic diagram of a glass slide placed on a stage according to an embodiment of the present disclosure.

[0012] Figure 5 It is a schematic diagram of the steps of a method for acquiring a focal length value according to an embodiment of the present disclosure.

[0013] Figure 6A AND Figure 6B It is a schematic diagram of a glass slide without deformation and with deformation according to an embodiment of the present disclosure.

[0014] Figure 7 It is a schematic diagram of the steps of obtaining a calibration slope value and a calibration intercept value according to an embodiment of the present disclosure.

[0015] SYMBOL DESCRIPTION:

[0016] 100: Focal length value acquisition device

[0017] 110: Body

[0018] 111: Side

[0019] 120: Stage

[0020] 121: Tabletop

[0021] 130: Linear movement module

[0022] 140: Probe

[0023] 150: Optical ruler module

[0024] 151: Graduated slide rail

[0025] 152: Optical scale reader / writer head

[0026] 160: Camera module

[0027] 170: Processor

[0028] 200: Slide

[0029] 210: Sampling slot

[0030] 220: Microscope shooting area

[0031] 230: Surface

[0032] 300: Cell sample Detailed implementation manners

[0033] Please refer to Figure 1 , Figure 1 which is a schematic diagram of the steps of a method for obtaining a focal length value according to an embodiment of the present disclosure, including: Step S100, obtaining a calibration slope value and a calibration intercept value of a detection device including a microscope; Step S200, obtaining a height value of a slide by determining that a probe contacts a scale of a stage and another scale of the slide according to an image of the stage; Step S300, calculating a calculated focal length of the slide according to the calibration slope value, the calibration intercept value and the height value. Specifically, in Step S300, the height value is substituted into the following formula to calculate the calculated focal length value of the slide: Ft = a×y + b, where Ft is the calculated focal length value of the slide, a is the calibration slope value, y is the height value, and b is the calibration intercept value.

[0034] Please also refer to Figure 2 and Figure 3 , Figure 1 The method for obtaining a focal length value can be achieved by a focal length value obtaining device 100 shown in Figure 2 and Figure 3 . In some embodiments, the focal length value obtaining device 100 includes a main body 110, a stage 120, a linear movement module 130, a probe 140, an optical scale module 150, a camera module 160, and a processor 170. The main body 110 includes a side surface 111. The stage 120 is located on one side of the side surface 111 and has a table surface 121 for placing the slide 200. The linear movement module 130 is connected to the main body 110. The probe 140 is moved relative to the side surface 111 closer to or farther from the stage 120 through the linear movement module 130. The processor 170 is electrically connected to the linear movement module 130, the optical scale module 150, and the camera module 160.

[0035] In some embodiments, the optical scale module 150 includes a graduated slide rail 151 disposed on the side surface 111, and an optical scale reader / writer head 152 that moves along the graduated slide rail 151. The optical scale reader / writer head 152 is connected to the probe 140 and is configured to read the graduations of the graduated slide rail 151. As the probe 140 is displaced, the optical scale reader / writer head 152 can transmit the read graduations back to the processor 170.

[0036] In some embodiments, the imaging module 160 is configured to photograph the stage surface 121 and output an image. In some embodiments, the image output by the imaging module 160 to the processor 170 is a real-time image, which can be taken continuously or discontinuously. The field of view of the imaging module 160 includes the stage surface 121 of the stage 120 and the glass slide 200 placed on the stage surface 121. Specifically, the imaging module 160 should be able to clearly photograph the stage surface 121 of the stage 120 to provide a sufficiently clear real-time image for the processor 170 to determine whether the probe 140 contacts the stage 120, whether the probe 140 contacts the glass slide 200, and whether the glass slide 200 is deformed.

[0037] In some embodiments, the processor 170 is configured to perform the following operations: controlling the linear movement module 130 to move the probe 140 along the graduated slide rail 151 closer to or farther away from the stage 120; controlling the imaging module 160 to photograph a real-time image of the stage 120; calculating the height value of the glass slide 200 by determining the graduations when the probe 140 contacts the stage 120 and another graduation when the probe 140 contacts the glass slide 200 based on the real-time image of the stage 120; and calculating the calculated focal length value of the glass slide 200 based on the calibration slope value of the detection device including the microscope, the calibration intercept value of the detection device including the microscope, and the height value. Specifically, the processor 170 substitutes the height value into the formula Ft = a×y + b to calculate the calculated focal length value of the glass slide 200, where Ft is the calculated focal length value of the glass slide, a is the calibration slope value, y is the height value, and b is the calibration intercept value.

[0038] Please refer to Figure 4A and Figure 4B, in one embodiment of the present disclosure, the glass slide 200 used includes a sample injection groove 210 and a microscope photographing area 220. The cell sample 300 is injected into the glass slide 200 through the sample injection groove 210 and flows into the microscope photographing area 220. When performing detection, the microscope passes through the microscope photographing area 220 to photograph the cell sample 300 located on the surface 230. When the glass slide 200 is placed on the stage 120, the height difference between the surface 230 and the tabletop 121 is the height value of the glass slide 200. The manufacturing tolerance of the glass slide will cause slight differences in the height value of each glass slide, thereby affecting the focal length of the microscope focusing. In some embodiments, the method for obtaining the focal length value of the present disclosure further includes aligning the sample injection groove of the glass slide with the probe so that the probe can surely contact the surface of the glass slide for carrying the cell sample to ensure obtaining the correct height value.

[0039] Please refer to Figure 5 , Figure 5 for a more specific and detailed description of how to Figure 1 obtain the height value of the glass slide 200 through step S200 of the method by judging the scale where the probe contacts the stage and another scale where the probe contacts the glass slide. The following description should be read in conjunction with Figure 2 and Figure 3 the focal length value obtaining device 100.

[0040] The steps for obtaining the height value of the glass slide 200 include: step S210, moving the probe 140 from the initial position along the scale slide rail 151 towards the stage 120; step S220, when it is judged according to the real-time image of the stage 120 that the probe 140 contacts the stage 120, recording the scale of the probe 140 relative to the scale slide rail 151 as the reference scale; step S230, returning the probe 140 to the initial position and placing the glass slide 200 on the stage 120; step S240; when it is judged according to another real-time image of the stage 120 that the probe 140 contacts the glass slide 200 and deforms the glass slide 200, recording the other scale of the probe 140 relative to the scale slide rail 151 as the glass slide scale; and step S250, subtracting the glass slide scale from the reference scale to obtain the height value of the glass slide 200.

[0041] In conjunction with the above steps for obtaining the height value of the glass slide 200, the processor 170 is further configured to perform the following operations: when it is judged according to the real-time image that the probe 140 contacts the stage 120, reading the scale of the probe 140 relative to the scale slide rail 151 through the optical scale reader / writer 152 and recording it as the reference scale; when it is judged according to the real-time image that the probe 140 contacts the glass slide 200 and deforms the glass slide 200, reading the other scale of the probe 140 relative to the scale slide rail 151 through the optical scale reader / writer 152 and recording it as the glass slide scale; subtracting the glass slide scale from the reference scale to obtain the height value.

[0042] In some embodiments, when determining whether the glass slide is deformed, another real-time image is used to determine whether the surface of the glass slide carrying the cell sample is slightly deformed due to contact with the probe. Figure 6A As shown, when the probe 140 has not yet contacted the glass slide 200, the surface 230 of the glass slide 200 is slightly raised; Figure 6B As shown, the probe 140 has contacted the glass slide 200 and caused the glass slide 200 to deform. The surface 230 is pressed by the probe 140 and slightly deformed.

[0043] See also Figure 7 , Figure 7 The invention is a step of obtaining a correction slope value and a correction intercept value before executing the focal length acquisition method disclosed in the present invention, comprising: step S110, measuring the height value of a reference glass slide by the aforementioned measurement method for the glass slide height value; step S120, injecting a cell sample into the reference glass slide, and measuring the actual focal length value of the reference glass slide on a detection device including a microscope; step S130, repeating the step of measuring the actual focal length value of the reference glass slide at least five times; and step S140, substituting the reference height value and the actual focal length value into a formula Ft=a×y+b, and calculating the correction slope value and the correction intercept value of the detection device including the microscope, where Ft is the actual focal length value of the reference glass slide, a is the correction slope value, y is the height value, and b is the correction intercept value.

[0044] The following are the data and results of the experimental test of the focal length acquisition method and focal length acquisition device disclosed in the present invention. Through the above-mentioned focal length acquisition device, the focal length acquisition method disclosed in the present invention is used to calculate the height value and the calculated focal length value of different glass slides, and the actual focal length of these glass slides is actually measured by the microscope device for comparison. Since the cell image cannot be clearly captured as long as the focal length changes by more than 0.03 mm during microscope shooting, the calculated focal length value and the actual focal length value of the glass slide must differ by less than 0.03 mm to meet the accuracy requirements of microscope shooting.

[0045] Table 1 shows the data results of comparing the height values ​​and calculated focal lengths of the glass slides No. 1 to 5 by the focal length acquisition method and focal length acquisition device disclosed in the present invention, and the actual focal length values ​​measured by the No. 1 microscope device. The numerical units in Table 1 are millimeters.

[0046] [Table 1]

[0047]

[0048] Table 2 shows the data results of comparing the height values ​​and calculated focal lengths of the glass slides No. 1 to 5 by the focal length acquisition method and focal length acquisition device disclosed in the present invention, and the actual focal length values ​​measured by the second microscope device. The numerical units in Table 2 are millimeters.

[0049] [Table 2]

[0050]

[0051] According to Table 1 and Table 2, by means of the focal length value obtaining method and the focal length value obtaining device of the present disclosure, the error ranges between the calculated focal length values and the actual focal length values obtained are all less than 0.03 mm. In addition, although there are slight differences in the focal length parameters of each microscope, by means of the focal length value obtaining method and the focal length value obtaining device of the present disclosure, even on different microscope devices, the accuracy condition that the error ranges between the calculated focal length values and the actual focal length values are all less than 0.03 mm can still be satisfied. The focal length value obtaining method and the focal length value obtaining device of the present disclosure can overcome the differences in the focal length parameters of different microscope devices.

[0052] In some embodiments, the focal length value obtaining method further includes generating an information label for calculating the focal length of the slide. In some embodiments, the processor of the focal length value obtaining device is further configured to generate an information label for calculating the focal length of the slide. In some embodiments, the information label can be a one-dimensional barcode, a two-dimensional barcode (e.g., QR code), a radio-frequency identification (RFID), or other readable information label barcodes. The information label can be attached to the slide, and the calculated focal length of the slide can be directly obtained by scanning the information label, without the need for re-measurement, so as to improve the detection efficiency.

[0053] The present disclosure also relates to a detection device including a focal length value obtaining device for detecting a biological sample located on a slide. The detection device including the focal length value obtaining device includes the focal length value obtaining device and a microscope module as described above. In some embodiments as described above, the detection device including the focal length value obtaining device is a circulating tumor cell detection (CTC) system. The calculated focal length value of the slide carrying the cell sample is obtained by the focal length value obtaining device, and then the microscope module focuses and takes a picture according to the calculated focal length value to obtain a clear sample image for result interpretation.

[0054] In summary, the focal length value obtaining method, the focal length value obtaining device, and the detection device including the focal length value obtaining device of the present disclosure accurately measure the height value of the slide by identifying the deformation of the slide through an image, and obtain an accurate focal length value through formula conversion. Therefore, the automatic focusing technology can be realized, the problem of tiny focusing errors in the microscope can be overcome, the shooting speed and accuracy can be improved to take clear photos, and thus the efficiency of automatic detection can be improved.

Claims

1. A method for obtaining a focal length value, comprising: Obtaining a calibration slope value and a calibration intercept value of a detection device including a microscope; According to the plurality of images of the stage, the height value of the glass slide is obtained by determining the scale mark at which the probe contacts the stage and another scale mark at which the probe contacts the glass slide; and The calculated focal length value of the glass slide is calculated according to the corrected slope value, the corrected intercept value and the height value.

2. The method for obtaining focal length value according to claim 1, characterized in that: Obtaining the height value further includes: Moving the probe along the scale slide toward the carrier; When it is determined that the probe contacts the stage according to the images related to the stage, recording the scale of the probe relative to the scale rail as a reference scale; placing the glass slide on the stage; When it is determined according to the images of the stage that the probe contacts the glass slide and causes the glass slide to deform, the other scale of the probe relative to the scale rail is recorded as the glass slide scale; Subtract the slide scale from the reference scale to obtain the height value.

3. The method for obtaining focal length value according to claim 2, characterized in that: The deformation of the glass slide is determined by judging, based on the images, that the surface of the glass slide carrying the cell sample is deformed due to contact with the probe.

4. The method for obtaining focal length value according to claim 1, characterized in that: The method further comprises aligning the sample injection slot of the glass slide with the probe.

5. The method for obtaining focal length value according to claim 1, characterized in that: The method further includes generating an information tag including the calculated focal length of the slide.

6. A focal length value acquisition device, comprising: The body, including the sides; A stage, located on one side of the side, having a table surface for placing a glass slide; A linear moving module connected to the main body; A probe, moving closer to or farther from the carrier relative to the side surface through the linear movement module; An optical ruler module, comprising a scale rail disposed on the side, and an optical ruler read / write head moving along the scale rail, wherein the optical ruler read / write head is connected to the probe and configured to read the scale of the scale rail; Camera module; as well as A processor configured to perform the following operations: Control the linear motion module to move the probe along the scale slide rail toward or away from the carrier; Controlling the camera module to capture multiple images on the stage; Calculating the height of the glass slide by determining the scale mark at which the probe contacts the glass slide and another scale mark at which the probe contacts the glass slide according to the images of the stage; as well as The calculated focal length value of the glass slide is calculated according to the calibrated slope value of the detection device including the microscope, the calibrated intercept value of the detection device including the microscope and the height value.

7. The focal length value acquisition device according to claim 6, characterized in that: The processor is further configured to perform the following operations: When it is determined according to the images that the probe contacts the stage, the scale of the probe relative to the scale rail is read by the optical ruler reading and writing head, and recorded as a reference scale; When it is determined according to the images that the probe contacts the glass slide and causes the glass slide to deform, the other scale of the probe relative to the scale rail is read by the optical ruler read-write head and recorded as the glass slide scale; Subtract the slide scale from the reference scale to obtain the height value.

8. The focal length value acquisition device according to claim 6, characterized in that: The deformation of the glass slide refers to that the surface of the glass slide carrying the cell sample is deformed due to contact with the probe.

9. The focal length value acquisition device according to claim 6, characterized in that: The processor is further configured to generate an information tag including the calculated focal length of the slide.

10. A detection device capable of calculating the focal length of a glass slide, used for detecting a biological sample located on a glass slide, the detection device comprising: The focal length value acquisition device as claimed in any one of claims 6 to 9, configured to obtain the calculated focal length value of the slide; and The microscope module is configured to focus and photograph according to the calculated focal length value.

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