TWI733355B - Pipetting system and calibration method thereof - Google Patents
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Abstract
Description
本發明是有關於移液設備技術領域,且特別是有關於一種移液系統及其校準方法。The present invention relates to the technical field of pipetting equipment, and particularly relates to a pipetting system and a calibration method thereof.
液體的抽取與注射是產業應用中經常需要進行的操作。在自動化過程中,可利用移液設備進行液體的抽取與注射,通過移液設備的移液管和轉盤達到微量移液的操作要求。Liquid extraction and injection are operations that are often required in industrial applications. In the automated process, the pipetting equipment can be used for liquid extraction and injection, and the pipette and turntable of the pipetting equipment can meet the operation requirements of micropipetting.
實際應用中,不同液體、不同操作場景均需要換用不同的移液管。在移液管的拆裝過程中,由於組裝偏差、轉盤的移動偏差等原因,造成移液管的尖端位置產生三個維度不同程度的偏移。這種偏移對於微量移液操作造成如下影響:In practical applications, different liquids and different operating scenarios need to be replaced with different pipettes. In the process of disassembling and assembling the pipette, due to the assembly deviation, the movement deviation of the turntable, etc., the tip position of the pipette is shifted in three dimensions to varying degrees. This shift has the following effects on micropipetting operations:
在抽取液體時,移液管尖端的位置偏移造成移液管吸入氣泡,過多的氣泡會造成後續操作的錯誤;When extracting liquid, the offset of the tip of the pipette will cause the pipette to inhale air bubbles. Excessive air bubbles will cause errors in subsequent operations;
在注射液體時,移液管尖端的位置偏移造成液體無法順利注射至預定位置,導致液體流失;以及When injecting liquid, the displacement of the tip of the pipette causes the liquid to be unable to be injected to the predetermined position smoothly, resulting in liquid loss;
在轉盤轉動時,移液管尖端的位置偏移造成移液管與轉盤上的試管等部件發生碰撞,影響系統運行。When the turntable rotates, the position deviation of the pipette tip causes the pipette to collide with the test tube on the turntable and other parts, which affects the operation of the system.
可見,由於移液管尖端的位置偏移,移液操作和系統運行受到影響。It can be seen that due to the offset of the pipette tip, the pipetting operation and system operation are affected.
需要說明的是,上述背景技術部分公開的資訊僅用於加強對本案的背景的理解,因此可以包括不構成對本領域具有通常知識者已知的現有技術的資訊。It should be noted that the information disclosed in the background art section above is only used to enhance the understanding of the background of the case, and therefore may include information that does not constitute the prior art known to those with ordinary knowledge in the field.
有鑑於此,本案揭露一種移液系統及其校準方法,以實現移液系統的校準,克服現有技術中由於移液管尖端的位置偏移導致移液操作和系統運行受到影響的問題。In view of this, this case discloses a pipetting system and a calibration method thereof to achieve the pipetting system calibration and overcome the problem that the pipetting operation and system operation are affected due to the position offset of the pipette tip in the prior art.
根據本發明之一方面,提出一種移液系統的校準方法,所述校準方法包括:通過一影像擷取裝置擷取一校準點的至少一第一影像;獲得影像擷取裝置的一中心點相對於第一影像中的校準點於一平面的一第一偏移量,依據第一偏移量調整影像擷取裝置的位置,使所述中心點與所述校準點進行校準;通過校準後的影像擷取裝置擷取移液系統的一移液管的尖端的至少一第二影像和一第三影像,其中所述第三影像擷取自所述尖端與校準後的所述中心點的對焦距離發生位移之後;獲得第三影像中的所述尖端相對於第二影像中的所述尖端於所述平面的一第二偏移量;分別依據位移、第三影像相對於第二影像的清晰度值的變異量和一預設的對焦距離與清晰度值的映射關係表,獲得第三影像的清晰度值相對於一預設清晰度值的一對焦距離偏移量;以及,依據第二偏移量和所述對焦距離偏移量調整移液管的位置,使所述尖端與校準後的所述中心點進行校準。According to one aspect of the present invention, a method for calibrating a pipetting system is provided. The calibration method includes: capturing at least one first image of a calibration point through an image capturing device; A first offset of the calibration point in the first image on a plane, and the position of the image capturing device is adjusted according to the first offset, so that the center point and the calibration point are calibrated; The image capture device captures at least a second image and a third image of the tip of a pipette of the pipetting system, wherein the third image is captured from the focus of the tip and the center point after calibration After the distance is shifted; obtain a second offset of the tip in the third image relative to the tip in the second image on the plane; respectively according to the displacement and the sharpness of the third image relative to the second image The degree of variation of the degree value and a preset mapping relationship table between the focus distance and the sharpness value to obtain a focusing distance offset of the sharpness value of the third image with respect to a predetermined sharpness value; and, according to the second The offset and the focus distance offset adjust the position of the pipette so that the tip is calibrated with the center point after calibration.
在本案的一個實施例中,獲得影像擷取裝置的一中心點相對於所述第一影像中的所述校準點於一平面的一第一偏移量的步驟包括:分別獲得所述中心點和所述第一影像中的所述校準點於所述平面的直角坐標,其中所述平面平行於所述移液系統的一轉盤所在的平面;分別將所述中心點的直角坐標和所述校準點的直角坐標轉換為極角,以獲得所述中心點的極角相對於所述校準點的極角的夾角;以及,依據所述夾角獲得所述第一偏移量。In an embodiment of the present case, the step of obtaining a first offset of a center point of the image capturing device relative to the calibration point in the first image on a plane includes: obtaining the center points respectively And the Cartesian coordinates of the calibration point in the first image on the plane, wherein the plane is parallel to the plane where a turntable of the pipetting system is located; and the Cartesian coordinates of the center point and the The rectangular coordinate of the calibration point is converted into a polar angle to obtain the included angle of the polar angle of the center point with respect to the polar angle of the calibration point; and the first offset is obtained according to the included angle.
在本案的一個實施例中,所述獲得所述第一影像中的所述校準點於所述平面的直角坐標的步驟包括:對所述第一影像進行平滑濾波,去除所述第一影像的噪點;對平滑濾波後的所述第一影像進行二值化,為平滑濾波後的所述第一影像中的每個圖元點賦值;對二值化後的所述第一影像進行測邊,標示出二值化後的所述第一影像的多個邊緣點;對測邊後的所述第一影像的所述多個邊緣點進行輪廓標定,形成至少一個輪廓;對各個所述輪廓進行圓度檢測,篩除圓度值小於一預設值的輪廓;對篩除後的所述輪廓進行面積篩選,篩出輪廓面積符合所述校準點的面積的一輪廓;以及,獲得篩出的所述輪廓的形心於所述平面的直角坐標,作為所述第一影像中的所述校準點於所述平面的直角坐標。In an embodiment of the present case, the step of obtaining the rectangular coordinates of the calibration point in the first image on the plane includes: smoothing and filtering the first image to remove Noise; binarize the first image after smoothing and filtering, and assign a value to each pixel point in the first image after smoothing and filtering; perform edge measurement on the first image after binarization , Marking a plurality of edge points of the first image after binarization; performing contour calibration on the plurality of edge points of the first image after edge measurement to form at least one contour; for each of the contours Perform roundness detection to screen out contours whose roundness value is less than a preset value; perform area screening on the screened contours, and screen out a contour whose area meets the area of the calibration point; and obtain a screened out contour The rectangular coordinates of the centroid of the contour on the plane are used as the rectangular coordinates of the calibration point in the first image on the plane.
在本案的一個實施例中,獲得所述第三影像中的所述尖端相對於所述第二影像中的所述尖端於一平面的一第二偏移量的步驟包括:分別獲得所述第三影像中的所述尖端的中心於所述平面的一第一直角坐標,和所述第二影像中的所述尖端的中心於所述平面的一第二直角坐標,其中所述平面垂直於所述對焦距離所在的方向;依據所述第一直角坐標和所述第二直角坐標,獲得所述第三影像中的所述尖端的中心相對於所述第二影像中的所述尖端的中心的圖元偏移量,其中所述直角坐標以一圖元作為一座標單位;以及,依據一圖元與距離的對應關係,將所述圖元偏移量轉換成距離偏移量,作為所述第二偏移量。In an embodiment of the present case, the step of obtaining a second offset of the tip in the third image relative to the tip in the second image on a plane includes: obtaining the first The center of the tip in the three images is a first rectangular coordinate of the plane, and the center of the tip in the second image is a second rectangular coordinate of the plane, wherein the plane is perpendicular to The direction in which the focusing distance is located; according to the first rectangular coordinates and the second rectangular coordinates, the center of the tip in the third image is obtained relative to the center of the tip in the second image , Where the rectangular coordinate uses a graphic element as a standard unit; and, according to the corresponding relationship between a graphic element and a distance, the graphic element offset is converted into a distance offset, which is used as the The second offset.
在本案的一個實施例中,所述獲得所述第三影像中的所述尖端的中心於所述平面的一第一直角坐標的步驟包括:對所述第三影像進行平滑濾波,去除所述第三影像的噪點;對平滑濾波後的所述第三影像進行二值化,為平滑濾波後的所述第三影像中的每個圖元點賦值;對二值化後的所述第三影像進行測邊,標示出二值化後的所述第三影像的多個邊緣點;對測邊後的所述第三影像的所述多個邊緣點進行輪廓標定,形成至少一個輪廓;對各個所述輪廓進行圓度檢測,篩除圓度值小於一預設值的輪廓;對篩除後的所述輪廓進行面積篩選,篩出輪廓面積符合所述尖端的面積的一輪廓;以及,獲得篩出的所述輪廓的形心於所述平面的直角坐標,作為所述第三影像中的所述尖端的中心於所述平面的直角坐標。In an embodiment of the present case, the step of obtaining a first right-angle coordinate of the center of the tip in the third image on the plane includes: smoothly filtering the third image to remove the The noise of the third image; binarize the third image after smoothing and filtering, and assign a value to each pixel point in the third image after smoothing and filtering; Image edge measurement, marking multiple edge points of the third image after binarization; performing contour calibration on the multiple edge points of the third image after edge measurement to form at least one contour; Perform roundness detection on each of the contours, and screen out contours whose roundness value is less than a preset value; perform area screening on the screened contours, and screen out a contour whose area matches the area of the tip; and, Obtain the rectangular coordinates of the centroid of the screened contour on the plane as the rectangular coordinates of the center of the tip in the third image on the plane.
在本案的一個實施例中,所述依據一圖元與距離的對應關係的步驟包括:通過所述影像擷取裝置擷取一預設尺寸的物體的影像;以及,根據所述物體的影像的圖元個數與所述預設尺寸,獲得所述圖元與距離的對應關係。In an embodiment of the present case, the step of according to the correspondence between a picture element and the distance includes: capturing an image of an object of a preset size through the image capturing device; and, according to the image of the object The number of graphic elements and the preset size are used to obtain the corresponding relationship between the graphic elements and the distance.
在本案的一個實施例中,所述分別依據所述位移、所述第三影像相對於所述第二影像的清晰度值的變異量和一預設的對焦距離與清晰度值的映射關係表,獲得所述第三影像的清晰度值相對於一預設清晰度值的一對焦距離偏移量的步驟包括:依據所述位移的方向和所述第三影像相對於所述第二影像的清晰度值的變異量,獲得所述第三影像的清晰度值在所述映射關係表中的位置,其中所述映射關係表中所述清晰度值呈高斯分佈;以及,根據所述第三影像的清晰度值在所述映射關係表中的位置,獲得所述第三影像的清晰度值對應的對焦距離相對於一預設清晰度值對應的對焦距離的所述對焦距離偏移量,其中所述預設清晰度值是所述映射關係表中的最高清晰度值。In an embodiment of the present case, the mapping relationship between the focus distance and the sharpness value is based on the displacement, the deviation of the sharpness value of the third image relative to the second image, and a preset focusing distance and sharpness value. , The step of obtaining a focus distance offset of the sharpness value of the third image relative to a preset sharpness value includes: according to the direction of the displacement and the third image relative to the second image Obtaining the position of the sharpness value of the third image in the mapping relationship table, wherein the sharpness value in the mapping relationship table has a Gaussian distribution; and, according to the variation amount of the sharpness value, the position of the sharpness value of the third image in the mapping relationship table is obtained; The position of the sharpness value of the image in the mapping relationship table, obtaining the focusing distance offset of the focusing distance corresponding to the sharpness value of the third image with respect to the focusing distance corresponding to a preset sharpness value, The preset sharpness value is the highest sharpness value in the mapping relationship table.
在本案的一個實施例中,所述校準方法還包括設置所述影像擷取裝置的成像景深,使所述對焦距離偏移量的可偵測範圍位於一預設距離範圍內,其中,擷取所述第三影像的步驟中,所述位移大於所述預設距離範圍。In an embodiment of the present case, the calibration method further includes setting the imaging depth of the image capture device so that the detectable range of the focus distance offset is within a preset distance range, wherein In the step of the third image, the displacement is greater than the preset distance range.
根據本發明之一方面,提出一種移液系統,所述移液系統包括:一影像擷取裝置;以及一處理器,所述處理器被配置為通過執行多條可執行指令實現:通過所述影像擷取裝置擷取一校準點的至少一第一影像;獲得所述影像擷取裝置的一中心點相對於所述第一影像中的所述校準點於一平面的一第一偏移量,依據所述第一偏移量調整所述影像擷取裝置的位置,使所述中心點與所述校準點進行校準;通過校準後的所述影像擷取裝置擷取所述移液系統的一移液管的尖端的至少一第二影像和一第三影像,其中所述第三影像擷取自所述尖端與校準後的所述中心點的對焦距離發生位移之後;獲得所述第三影像中的所述尖端相對於所述第二影像中的所述尖端於所述平面的一第二偏移量;分別依據所述位移、所述第三影像相對於所述第二影像的清晰度值的變異量和一預設的對焦距離與清晰度值的映射關係表,獲得所述第三影像的清晰度值相對於一預設清晰度值的一對焦距離偏移量;以及,依據所述第二偏移量和所述對焦距離偏移量調整所述移液管的位置,使所述尖端與校準後的所述中心點進行校準。According to one aspect of the present invention, a pipetting system is provided. The pipetting system includes: an image capture device; and a processor configured to execute a plurality of executable instructions: The image capturing device captures at least one first image of a calibration point; obtaining a first offset of a center point of the image capturing device relative to the calibration point in the first image on a plane , Adjust the position of the image capturing device according to the first offset, so that the center point and the calibration point are calibrated; and the image capturing device is calibrated to capture the pipetting system At least a second image and a third image of the tip of a pipette, wherein the third image is captured after the focus distance between the tip and the calibrated center point is shifted; the third image is obtained A second offset of the tip in the image relative to the tip in the second image relative to the plane; respectively according to the displacement and the sharpness of the third image relative to the second image The degree value variation and a preset mapping relationship table between the focus distance and the sharpness value to obtain a focusing distance offset of the sharpness value of the third image relative to a preset sharpness value; and, according to The second offset and the focus distance offset adjust the position of the pipette so that the tip is calibrated with the center point after calibration.
在本案的一個實施例中,所述移液系統還包括:一具有所述影像擷取裝置的轉盤。In an embodiment of the present case, the pipetting system further includes: a turntable with the image capturing device.
本案與現有技術相比的有益效果至少包括:Compared with the prior art, the beneficial effects of this case include at least:
通過影像擷取裝置擷取校準點的影像,獲得影像擷取裝置的中心點相對於校準點於平面維度的第一偏移量,實現影像擷取裝置於平面維度的校準,使校準後的影像擷取裝置用於後續校準步驟的定位;通過校準後的影像擷取裝置至少兩次擷取移液管的尖端的影像,獲得移液管的尖端於平面維度的第二偏移量以及於對焦距離維度的對焦距離偏移量,實現移液管的尖端於平面維度和對焦距離維度的校準;從而,本案利用了一影像擷取裝置實現移液系統的校準,從而實現後續移液操作和系統運行的控制。The image of the calibration point is captured by the image capture device, and the first offset of the center point of the image capture device relative to the calibration point in the plane dimension is obtained, so as to realize the calibration of the image capture device in the plane dimension, so that the calibrated image The capture device is used for the positioning of the subsequent calibration steps; the image capture device after the calibration is used to capture the image of the tip of the pipette at least twice to obtain the second offset of the tip of the pipette in the plane dimension and the focus The focus distance offset in the distance dimension realizes the calibration of the tip of the pipette in the plane dimension and the focus distance dimension; thus, this case uses an image capture device to achieve the pipetting system calibration, so as to realize the subsequent pipetting operation and system Operational control.
為了對本發明之上述及其他方面有更佳的瞭解,下文特舉實施例,並配合所附圖式詳細說明如下:In order to have a better understanding of the above and other aspects of the present invention, the following specific examples are given in conjunction with the accompanying drawings to describe in detail as follows:
現在將參考附圖更全面地描述示例實施方式。然而,示例實施方式能夠以多種形式實施,且不應被理解為限於在此闡述的實施方式。相反,提供這些實施方式使得本案將全面和完整,並將示例實施方式的構思全面地傳達給本領域的具有通常知識者。在圖中相同的附圖標記表示相同或類似的結構,因而將省略對它們的重複描述。Example embodiments will now be described more fully with reference to the accompanying drawings. However, the example embodiments can be implemented in various forms, and should not be construed as being limited to the embodiments set forth herein. On the contrary, these embodiments are provided so that the case will be comprehensive and complete, and the concept of the example embodiments will be fully conveyed to those with ordinary knowledge in the art. The same reference numerals in the figures indicate the same or similar structures, and thus their repeated description will be omitted.
第1圖示出一實施例中一種移液系統的主要結構,在一些實施例中,本案的移液系統的校準方法可用於校準如第1圖所示的移液系統。參照第1圖所示,該移液系統包括一轉盤10和一移液裝置30。移液裝置30例如是一個立式注射泵,通過移液管31實現移液操作。轉盤10上設置有移液管替換區101以及移液區102。Figure 1 shows the main structure of a pipetting system in an embodiment. In some embodiments, the method for calibrating the pipetting system in this case can be used to calibrate the pipetting system shown in Figure 1. Referring to FIG. 1, the pipetting system includes a
第2圖示出該移液系統中移液裝置30的主要結構,移液裝置30連接滑軌300,移液裝置30帶動移液管31沿滑軌300移動,實現移液操作。進一步的,本實施例的移液系統還包括一裝設於轉盤10上的影像擷取裝置21,以及一固定不動的校準點11。利用該影像擷取裝置21和該校準點11,實現移液系統的轉盤10和移液管31的校準。Figure 2 shows the main structure of the
第3圖示出一實施例中一移液系統的校準方法的步驟。參照第3圖所示,本實施例中移液系統的校準方法包括:步驟S10、通過一影像擷取裝置擷取一校準點的至少一第一影像;步驟S20、獲得該影像擷取裝置的一中心點相對於該第一影像中的該校準點於一平面的一第一偏移量,依據該第一偏移量調整該影像擷取裝置的位置,使該中心點與該校準點進行校準;步驟S30、通過校準後的該影像擷取裝置擷取該移液系統的一移液管的尖端的至少一第二影像和一第三影像,其中該第三影像擷取自該尖端與校準後的該中心點的對焦距離發生位移之後;步驟S40、獲得該第三影像中的該尖端相對於該第二影像中的該尖端於該平面的一第二偏移量;步驟S50、分別依據該位移、該第三影像相對於該第二影像的清晰度值的變異量和一預設的對焦距離與清晰度值的映射關係表,獲得該第三影像的清晰度值相對於一預設清晰度值的一對焦距離偏移量;以及步驟S60、依據該第二偏移量和該對焦距離偏移量調整該移液管的位置,使該尖端與校準後的該中心點進行校準。Figure 3 shows the steps of a method for calibrating a pipetting system in an embodiment. Referring to Figure 3, the calibration method of the pipetting system in this embodiment includes: step S10, capturing at least one first image of a calibration point through an image capturing device; step S20, obtaining the image capturing device A first offset of a center point relative to the calibration point in the first image on a plane, and the position of the image capturing device is adjusted according to the first offset so that the center point and the calibration point are aligned Calibration; step S30, capturing at least a second image and a third image of the tip of a pipette of the pipetting system by the image capturing device after calibration, wherein the third image is captured from the tip and After the calibrated focusing distance of the center point is shifted; step S40, obtaining a second offset of the tip in the third image relative to the tip in the second image on the plane; step S50, respectively According to the displacement, the variation of the sharpness value of the third image relative to the second image, and a preset mapping relationship table between the focus distance and the sharpness value, the sharpness value of the third image relative to a predetermined Set a focus distance offset of the sharpness value; and step S60, adjust the position of the pipette according to the second offset and the focus distance offset, so that the tip is calibrated with the center point after calibration .
本實施例通過影像擷取裝置擷取校準點的影像,獲得影像擷取裝置的中心點相對於校準點於平面維度的第一偏移量,實現影像擷取裝置於平面維度的校準,使校準後的影像擷取裝置用於後續校準步驟的定位;通過校準後的影像擷取裝置至少兩次擷取移液管的尖端的影像,獲得移液管的尖端於平面維度的第二偏移量以及於對焦距離維度的對焦距離偏移量,實現移液管的尖端於平面維度和對焦距離維度的校準;從而,本實施例利用了一影像擷取裝置實現移液系統的校準,從而實現後續移液操作和系統運行的控制。In this embodiment, the image of the calibration point is captured by the image capturing device, and the first offset of the center point of the image capturing device with respect to the calibration point in the plane dimension is obtained, so as to realize the calibration of the image capturing device in the plane dimension, so that the calibration The latter image capture device is used for positioning in subsequent calibration steps; the image capture device after calibration is used to capture images of the tip of the pipette at least twice to obtain the second offset of the tip of the pipette in the plane dimension And the focus distance offset in the focus distance dimension, to achieve the calibration of the tip of the pipette in the plane dimension and the focus distance dimension; thus, this embodiment uses an image capture device to achieve the calibration of the pipetting system, thereby achieving subsequent Control of pipetting operation and system operation.
其中,於本實施例中,影像擷取裝置是一個顯微鏡攝像頭,於其他實施例中,影像擷取裝置是一個微型攝像頭,只要其具備影像擷取功能即可。校準點是一個固定不動的參考點,於本實施例中,校準點是屬於移液系統,於其他實施例中,校準點不是屬於移液系統。Among them, in this embodiment, the image capturing device is a microscope camera. In other embodiments, the image capturing device is a miniature camera, as long as it has an image capturing function. The calibration point is a fixed reference point. In this embodiment, the calibration point belongs to the pipetting system. In other embodiments, the calibration point does not belong to the pipetting system.
第4圖示出一實施例中影像擷取裝置擷取校準點的第一影像的示意。參照第4圖所示,本實施例中校準點11位於影像擷取裝置21的上方,在步驟S10擷取校準點11的影像前,移動影像擷取裝置21至校準點11進入影像擷取裝置21的擷取區域內。在其他實施例中,校準點與影像擷取裝置之間的相對位置以調整,只要滿足校準點能進入影像擷取裝置的擷取區域內,使影像擷取裝置參照校準點進行校準的條件即可。FIG. 4 shows a schematic diagram of the image capturing device capturing the first image of the calibration point in an embodiment. Referring to Figure 4, the
第5圖示出一實施例中獲得一第一偏移量的主要步驟。參照第5圖所示,步驟S20中獲得該影像擷取裝置的一中心點相對於該第一影像中的該校準點於一平面的一第一偏移量的步驟包括:S202、分別獲得該中心點和該第一影像中的該校準點於該平面的直角坐標,其中該平面平行於該移液系統的一轉盤所在的平面;S204、分別將該中心點的直角坐標和該校準點的直角坐標轉換為極角,以獲得該中心點的極角相對於該校準點的極角的夾角;以及S206、依據該夾角獲得該第一偏移量。Figure 5 shows the main steps of obtaining a first offset in an embodiment. Referring to FIG. 5, the step of obtaining a first offset of a center point of the image capturing device relative to the calibration point in the first image on a plane in step S20 includes: S202, obtaining the The rectangular coordinates of the center point and the calibration point in the first image on the plane, wherein the plane is parallel to the plane where a turntable of the pipetting system is located; S204, the rectangular coordinates of the center point and the calibration point respectively The rectangular coordinate is converted into a polar angle to obtain the included angle of the polar angle of the center point with respect to the polar angle of the calibration point; and S206, obtaining the first offset amount according to the included angle.
結合第4圖和第5圖來看,該平面標示為X-Y平面,X-Y平面平行於影像擷取裝置21所在的平面。影像擷取裝置21所在的平面是移液系統中用於存放液體和接收液體的操作平臺,例如在配置轉盤的移液系統中,影像擷取裝置21所在的平面就是移液系統的轉盤。步驟S202中,先依據該X-Y平面建立X-Y直角坐標系,然後分別獲得中心點211和第一影像中的校準點11在該X-Y直角坐標系中的直角坐標。由於裝設完影像擷取裝置21後,中心點211的位置就是已知的,因此在一些實施例中,以中心點211為原點建立X-Y直角坐標系,並通過將第一影像投影至X-Y直角坐標系獲得第一影像中的校準點11的直角坐標,以方便第一偏移量的計算。在一些實施例中,預存一張中心點211的影像,並通過分別將中心點211的影像和第一影像投影至X-Y直角坐標系,獲得中心點211的直角坐標和校準點11的直角坐標。Combining Fig. 4 and Fig. 5, the plane is marked as the X-Y plane, and the X-Y plane is parallel to the plane where the
其中,校準點11的直角坐標是通過中心圓輪廓搜尋演算法獲得。第6圖示出一實施例中獲得校準點的直角坐標的主要步驟,參照第6圖所示,步驟S202中獲得該第一影像中的該校準點於該平面的直角坐標的步驟包括:S2021、對該第一影像進行平滑濾波,去除該第一影像的噪點;S2022、對平滑濾波後的該第一影像進行二值化,為平滑濾波後的該第一影像中的每個圖元點賦值;S2023、對二值化後的該第一影像進行測邊,標示出二值化後的該第一影像的多個邊緣點;S2024、對測邊後的該第一影像的該多個邊緣點進行輪廓標定,形成至少一個輪廓;S2025、對各個該輪廓進行圓度檢測,篩除圓度值小於一預設值的輪廓;S2026、對篩除後的該輪廓進行面積篩選,篩出輪廓面積符合該校準點的面積的一輪廓;以及S2027、獲得篩出的該輪廓的形心於該平面的直角坐標,作為該第一影像中的該校準點於該平面的直角坐標。Among them, the rectangular coordinates of the
具體來說,步驟S2022的二值化是將平滑濾波後的第一影像中超過一預設閾值的圖元點設為1,反之設為0。於本實施例中,預設閾值是根據需要設定,此處不作限定。步驟S2023的測邊是利用Canny邊緣檢測演算法將二值化後的第一影像的邊緣點標示出來。Canny邊緣檢測演算法是用來標識出 圖像中的實際邊緣的一個多級邊緣檢測演算法。步驟S2024的輪廓標定將上一步檢測到的相連的邊緣點連在一起形成輪廓,由於第一影像中可能存在多個輪廓,因此測邊時會檢測出多組相連的邊緣點,則輪廓標定時會標示出多個輪廓。步驟S2025的圓度檢測通過公式“T=4π*S/C
2”計算上一步標示出的各個輪廓的圓度值,並篩除圓度值低的輪廓,留下接近圓形的輪廓。用於評判圓度值的預設值是根據需要設定,此處不作限定。在公式“T=4π*S/C
2”中,T是一輪廓的圓度值,S是該輪廓的面積,C是該輪廓的周長。一輪廓的面積S和周長C是通過軟體測量獲得。校準點的面積是已知的,步驟S2026的面積篩選通過計算上一步留下的各個輪廓的面積,並篩出輪廓面積最接近校準點面積的一個輪廓,然後通過步驟S2027獲得該輪廓的形心於X-Y平面的直角坐標,作為校準點11的直角坐標。其中,一個輪廓的形心是該輪廓的幾何中心。
Specifically, the binarization in step S2022 is to set the pixel points in the first image after smoothing and filtering that exceed a preset threshold to 1, and vice versa. In this embodiment, the preset threshold is set according to needs, and is not limited here. The edge measurement in step S2023 is to use the Canny edge detection algorithm to mark the edge points of the first image after binarization. The Canny edge detection algorithm is a multi-level edge detection algorithm used to identify the actual edges in the image. The contour calibration of step S2024 connects the connected edge points detected in the previous step to form a contour. Since there may be multiple contours in the first image, multiple groups of connected edge points will be detected during edge measurement. Multiple outlines will be marked. The roundness detection in step S2025 calculates the roundness value of each contour marked in the previous step by the formula "T=4π*S/C 2 ", and screens out contours with low roundness values, leaving a contour close to a circle. The preset value used for judging the roundness value is set according to needs, and is not limited here. In the formula "T=4π*S/C 2 ", T is the roundness value of a contour, S is the area of the contour, and C is the circumference of the contour. The area S and perimeter C of a contour are obtained by software measurement. The area of the calibration point is known. The area screening of step S2026 calculates the area of each contour left in the previous step, and screens out a contour whose contour area is closest to the area of the calibration point, and then obtains the centroid of the contour through step S2027 The rectangular coordinates on the XY plane are used as the rectangular coordinates of the
同理,當需要獲取中心點211的直角坐標時,也採用中心圓輪廓搜尋演算法,此處不再展開說明。In the same way, when the rectangular coordinates of the
獲得中心點211的直角坐標和校準點11的直角坐標後,通過步驟S204分別將中心點211的直角坐標和校準點11的直角坐標轉換為極角。以將校準點11的直角坐標轉換為極角為例,通過下述公式獲得校準點11的極角θ
11:
After obtaining the rectangular coordinates of the
其中,(x
11,y
11)是校準點11於X-Y直角坐標系中的直角坐標,X-Y直角坐標系以一個圖元點作為單位座標。同理,根據中心點211的直角坐標(x
211,y
211)以獲得中心點211的極角θ
211,此處不再展開說明。進一步,將中心點211的極角θ
211減去校準點11的極角θ
11,獲得中心點211的極角θ
211相對於校準點11的極角θ
11的夾角Δθ,作為第一偏移量。在移液系統組態轉盤的實施例中,如第4圖所示,該夾角Δθ是轉盤沿其轉動方向的一個轉動角度。
Among them, (x 11 , y 11 ) are the rectangular coordinates of the
後續,依據該第一偏移量調整影像擷取裝置21的位置,使中心點211與校準點11進行校準,以補償第一偏移量。例如,控制移液系統的轉盤轉動Δθ,使中心點211沿垂直於X-Y平面的方向與校準點11對準。Subsequently, the position of the
第7圖示出一實施例中影像擷取裝置擷取一移液管的尖端的一第二影像的示意,第8圖示出一實施例中影像擷取裝置擷取移液管的尖端的一第三影像的示意。參照第7圖和第8圖所示,本實施例中移液管31位於影像擷取裝置21的上方,擷取移液管31的尖端311的影像前,移動移液管31至移液管31的尖端311進入影像擷取裝置21的擷取區域內。在其他實施例中,移液管與影像擷取裝置之間的相對位置可以調整,只要滿足移液管的尖端能進入影像擷取裝置的擷取區域內,並使移液管的尖端參照影像擷取裝置進行校準的條件。移液管31裝設於移液系統的一移液裝置30上,該移液裝置30例如一立式注射泵。其中,第7圖中擷取第二影像時,移液管31的尖端311的中心與影像擷取裝置21的中心點211在垂直於X-Y平面的Z坐標軸方向上是對準的,也即移液管31符合原始裝配的狀態,在原始裝配狀態下尖端311的中心投影至X-Y直角坐標系的原點。或者,以校準後的中心點211作為第7圖和第8圖中X-Y-Z三維坐標系的原點,由於中心點211已經相對於校準點11完成校準,將中心點211作為X-Y-Z三維坐標系的原點,有利於後續校準步驟的定位。第8圖中擷取第三影像時,移液管31重新裝設過,其尖端311與校準後的中心點211之間的對焦距離發生了一定的位移,並且尖端311與校準後的中心點211之間於X-Y平面也可能發生了一些位移。Figure 7 shows a schematic diagram of the image capturing device capturing a second image of the tip of a pipette in an embodiment, and Figure 8 shows the image capturing device capturing a second image of the tip of the pipette in an embodiment A schematic representation of the third image. Referring to Figures 7 and 8, the
第9圖示出一實施例中獲得一第二偏移量的主要步驟。參照第9圖所示,步驟S40中獲得該第三影像中的該尖端相對於該第二影像中的該尖端於該平面的一第二偏移量的步驟包括:S402、分別獲得該第三影像中的該尖端的中心於該平面的一第一直角坐標,和該第二影像中的該尖端的中心於該平面的一第二直角坐標,其中該平面垂直於該對焦距離所在的方向;S404、依據該第一直角坐標和該第二直角坐標,獲得該第三影像中的該尖端的中心相對於該第二影像中的該尖端的中心的圖元偏移量,其中該直角坐標以一圖元作為一座標單位;以及S406、依據一圖元與距離的對應關係,將該圖元偏移量轉換成距離偏移量,作為該第二偏移量。Figure 9 shows the main steps of obtaining a second offset in an embodiment. Referring to FIG. 9, the step of obtaining a second offset of the tip in the third image relative to the tip in the second image on the plane in step S40 includes: S402, obtaining the third image respectively A first rectangular coordinate of the center of the tip in the image on the plane, and a second rectangular coordinate of the center of the tip in the second image on the plane, wherein the plane is perpendicular to the direction of the focusing distance; S404. Obtain the pixel offset of the center of the tip in the third image relative to the center of the tip in the second image according to the first rectangular coordinate and the second rectangular coordinate, where the rectangular coordinate is A graphic element is used as a standard unit; and S406, according to the corresponding relationship between a graphic element and the distance, the graphic element offset is converted into a distance offset as the second offset.
結合第7圖至第9圖來看,對焦距離所在的方向是Z軸方向,第二偏移量是第三影像中的尖端311的中心相對於第二影像中的尖端311的中心,也即校準後的中心點211於X-Y平面的偏移量Δx和Δy。由於X-Y直角坐標系以一個圖元點作為單位座標,因此獲得的座標偏移量Δx和Δy就是圖元偏移量。進一步,依據圖元與距離的對應關係,即可獲得圖元偏移量所對應的距離偏移量。In combination with Figures 7-9, the direction of the focus distance is the Z-axis direction, and the second offset is the center of the
步驟S402中獲得第三影像中的尖端的中心的第一直角坐標是採用中心圓輪廓搜尋演算法。第10圖示出一實施例中獲得第三影像中的尖端的中心的一第一直角坐標的主要步驟,參照第10圖所示,步驟S402中獲得該第三影像中的該尖端的中心於該平面的一第一直角坐標的步驟包括:S4021、對該第三影像進行平滑濾波,去除該第三影像的噪點;S4022、對平滑濾波後的該第三影像進行二值化,為平滑濾波後的該第三影像中的每個圖元點賦值;S4023、對二值化後的該第三影像進行測邊,標示出二值化後的該第三影像的多個邊緣點;S4024、對測邊後的該第三影像的該多個邊緣點進行輪廓標定,形成至少一個輪廓;S4025、對各個該輪廓進行圓度檢測,篩除圓度值小於一預設值的輪廓;S4026、對篩除後的該輪廓進行面積篩選,篩出輪廓面積符合該尖端的面積的一輪廓;以及S4027、獲得篩出的該輪廓的圓心於該平面的直角坐標,作為該第三影像中的該尖端的中心於該平面的直角坐標。中心圓輪廓搜尋演算法的各個步驟的具體原理與上述實施例中校準點11的直角坐標的獲取過程同理,因此不再重複說明。In step S402, the first right-angle coordinate of the center of the tip in the third image is obtained by using a center circle contour search algorithm. Figure 10 shows the main steps of obtaining a first right-angle coordinate of the center of the tip in the third image in an embodiment. Referring to Figure 10, the center of the tip in the third image is obtained in step S402. The step of a first rectangular coordinate of the plane includes: S4021, smoothing and filtering the third image to remove the noise of the third image; S4022, binarizing the smoothed and filtered third image, which is a smoothing filter Assign a value to each pixel point in the third image afterwards; S4023, perform edge measurement on the third image after binarization, and mark multiple edge points of the third image after binarization; S4024, Perform contour calibration on the plurality of edge points of the third image after edge measurement to form at least one contour; S4025. Perform roundness detection on each contour, and screen out contours whose roundness value is less than a preset value; S4026, Perform area screening on the screened contour, and screen out a contour whose contour area matches the area of the tip; and S4027. Obtain the rectangular coordinates of the center of the screened contour on the plane as the third image. The center of the tip is in the rectangular coordinates of the plane. The specific principle of each step of the center circle contour search algorithm is the same as the process of obtaining the rectangular coordinates of the
如上所述,在一些實施例中,第二影像中的尖端311的中心於X-Y平面的第二直角坐標是X-Y直角坐標系的原點。在另一些實施例中,當需要獲取第二影像中的尖端311的中心於X-Y平面的第二直角坐標時,同樣採用中心圓輪廓搜尋演算法,此處不再展開說明。As described above, in some embodiments, the second rectangular coordinate of the center of the
步驟S406通過影像擷取裝置21拍攝一已知尺寸的物體,來獲得圖元與距離的對應關係。第11圖示出一實施例中獲得一圖元與距離的對應關係的主要步驟,參照第11圖所示,步驟S406中依據一圖元與距離的對應關係的步驟包括:S4062、通過該影像擷取裝置擷取一預設尺寸的物體的影像;以及S4064、根據該物體的影像的圖元個數與該預設尺寸,獲得該圖元與距離的對應關係。例如,通過影像擷取裝置21擷取一已知為100μm長的物體的影像,得到該物體的影像的圖元個數為10個,則獲得1圖元等於10μm的對應關係。根據該圖元與距離的對應關係,將第三影像中的尖端311相對於第二影像中的尖端311的圖元偏移量轉換成偏移距離。In step S406, an object of a known size is captured by the
例如,在一個示例中,參照第7圖和第8圖所示,通過中心圓輪廓搜尋演算法計算得到第二影像中的尖端311的中心於X-Y平面的第二直角坐標是(297,249),第三影像中的尖端311的中心於X-Y平面的第一直角坐標是(240,257)。則獲得第三影像中的尖端311於X坐標軸的偏移量Δx=-57,於Y坐標軸的偏移量Δy=8。進一步根據圖元與距離的對應關係,以獲得第三影像中的尖端311在X軸偏移了570μm,在Y軸偏移了80μm。For example, in an example, referring to Figures 7 and 8, the second rectangular coordinates of the center of the
第二影像和第三影像還用於獲得尖端311的對焦距離偏移量。第12圖示出一實施例中獲得對焦距離偏移量的主要步驟,參照第12圖所示,本實施例中,步驟S50中分別依據該位移、該第三影像相對於該第二影像的清晰度值的變異量和一預設的對焦距離與清晰度值的映射關係表,獲得該第三影像的清晰度值相對於一預設清晰度值的一對焦距離偏移量的步驟包括:S502、依據該位移的方向和該第三影像相對於該第二影像的清晰度值的變異量,獲得該第三影像的清晰度值在該映射關係表中的位置,其中該映射關係表中該清晰度值呈高斯分佈;以及S504、根據該第三影像的清晰度值在該映射關係表中的位置,獲得該第三影像的清晰度值對應的對焦距離相對於一預設清晰度值對應的對焦距離的該對焦距離偏移量,其中該預設清晰度值是該映射關係表中的最高清晰度值。The second image and the third image are also used to obtain the focus distance offset of the
其中,對焦距離與清晰度值的映射關係表是預先設定的。在進行移液系統的校準之前,先通過影像擷取裝置多次拍攝尖端的影像,計算不同對焦距離下的清晰度值,獲得能夠反應對焦距離與清晰度值之間的對應關係的映射關係表。例如,在一個實施例中,對焦距離與清晰度值的映射關係表如下表1所示:Among them, the mapping relationship table between the focus distance and the sharpness value is preset. Before calibrating the pipetting system, first use the image capture device to shoot the cutting-edge images multiple times, calculate the sharpness value at different focusing distances, and obtain a mapping table that can reflect the correspondence between the focusing distance and the sharpness value. . For example, in an embodiment, the mapping relationship between the focus distance and the sharpness value is shown in Table 1 below:
表1:對焦距離與清晰度值的映射關係表:
其中,清晰度值是正規化後的清晰度值。由於尖端與影像擷取裝置的中心點之間存在一個最優對焦距離,當尖端與中心點之間的距離等於該最優對焦距離時,影像擷取裝置所拍攝的尖端的影像具有最高清晰度值。當尖端與中心點之間的距離相對於該最優對焦距離變大或變小時,影像擷取裝置所拍攝的尖端的影像的清晰度值都會相對於該最高清晰度值減小。也就是說,影像的清晰度值關於對焦距離呈現高斯分佈(Gaussian distribution)的特徵。在該規律下,所獲取到的映射關係表中的清晰度值呈高斯分佈。因此,在獲得尖端與中心點之間的對焦距離時,需要至少兩次拍攝對焦距離發生了位移的尖端的影像,以獲得一清晰度值在映射關係表中的具體位置。在一些實施例中,多次重新裝設移液管,直至能夠獲取到尖端與中心點之間的準確對焦距離。需要說明的是,於本實施例中,上述映射關係表中僅展示出了部分對焦距離和清晰度值,於其他實施例中,在映射關係表中,最高清晰度值左右兩側的多個清晰度值大致是對稱分佈的。Among them, the sharpness value is the sharpness value after normalization. Since there is an optimal focusing distance between the tip and the center point of the image capturing device, when the distance between the tip and the center point is equal to the optimal focusing distance, the sharp image captured by the image capturing device has the highest definition value. When the distance between the tip and the center point becomes larger or smaller with respect to the optimal focusing distance, the sharpness value of the sharp image captured by the image capturing device will decrease relative to the highest sharpness value. In other words, the sharpness value of the image has a Gaussian distribution with respect to the focus distance. Under this rule, the definition value in the acquired mapping table is Gaussian. Therefore, when obtaining the focus distance between the tip and the center point, it is necessary to shoot the image of the tip with the shifted focus distance at least twice to obtain a specific position of the sharpness value in the mapping relationship table. In some embodiments, the pipette is reinstalled multiple times until the exact focusing distance between the tip and the center point can be obtained. It should be noted that in this embodiment, only part of the focus distance and sharpness values are shown in the above mapping relationship table. In other embodiments, in the mapping relationship table, there are multiple left and right sides of the highest sharpness value. The sharpness value is approximately symmetrically distributed.
參照第7圖和第8圖所示,位移是一個沿Z坐標軸方向的已知量。通過使尖端311與中心點211之間的對焦距離發生位移,使兩次擷取的尖端311的影像的清晰度值發生變化,以供獲得尖端311的第三影像的清晰度值對應的對焦距離,作為尖端311與中心點211之間的實際距離。以第7圖和第8圖所示的Z坐標軸的方向為例,當尖端311相對於影像擷取裝置21的中心點211沿Z坐標軸上移了一定距離,則尖端311與校準後的中心點211之間的對焦距離增大;當尖端311相對於影像擷取裝置21的中心點211沿Z坐標軸下移了一定距離,則尖端311與校準後的中心點211之間的對焦距離減小。Referring to Figures 7 and 8, the displacement is a known quantity along the Z coordinate axis. By shifting the focus distance between the
第三影像相對於第二影像的清晰度值的變異量是指第三影像的清晰度值相對於第二影像的清晰度值的差值。採用已有的方式獲得影像的清晰度值。具體來說,影像的清晰度值的評價演算法有多種,在空間域中,主要是考察影像的空間上的對比度,即相鄰圖元間的灰度特徵的梯度差;在頻域中,主要是考察影像的頻率分量,對焦清晰的影像高頻分量較多,對焦模糊的影像低頻分量較多。在一個實施例中,採用Laplace(拉普拉斯)運算元獲得第二影像和第三影像的清晰度值,Laplace運算元分別計算X軸方向和Y軸方向的梯度,同一場景下影像越清晰,梯度值越高。Laplace運算元的公式如下: 其中,Src指的是原始圖像的二維矩陣,而dst就是以src分別對x及y作二階偏微再加總的二維矩陣,目的是藉由求x方向與y方向的變化量找出影像的邊緣值,也就是清晰度值。 The amount of variation of the sharpness value of the third image with respect to the second image refers to the difference between the sharpness value of the third image and the sharpness value of the second image. Use the existing method to obtain the sharpness value of the image. Specifically, there are many algorithms for evaluating the sharpness value of an image. In the spatial domain, it is mainly to examine the spatial contrast of the image, that is, the gradient difference of gray-scale features between adjacent pixels; in the frequency domain, It mainly examines the frequency components of the image. The image with sharp focus has more high frequency components, and the image with blurred focus has more low frequency components. In one embodiment, the Laplace operator is used to obtain the sharpness values of the second image and the third image, and the Laplace operator calculates the gradients in the X-axis direction and the Y-axis direction respectively. The clearer the image in the same scene , The higher the gradient value. The formula of the Laplace operator is as follows: Among them, Src refers to the two-dimensional matrix of the original image, and dst is the two-dimensional matrix obtained by subtracting x and y respectively with src, and then adding up the two-dimensional matrix. The purpose is to find the amount of change in the x and y directions. The edge value of the output image, that is, the sharpness value.
在一個實施例中,預設清晰度值為上述映射關係表中的最高清晰度值1,其對應的最優對焦距離為6280μm。根據Laplace運算元計算得到尖端的第二影像的清晰度值為0.865822,然後增大對焦距離,將移液管上移20μm,再次拍攝獲得尖端的第三影像,通過Laplace運算元計算得到第三影像的清晰度值為0.890014。根據隨對焦距離增大而清晰度值增大的特徵和高斯分佈的規律,可知相對於最優對焦距離,拍攝第二影像和第三影像時尖端與中心點之間的對焦距離都小於該最優對焦距離,則確定在映射關係表中第三影像的清晰度值0.890014位於最高清晰度值1的左側,進而獲得第三影像的清晰度值0.890014所對應的對焦距離為6260μm。當然,在移動移液管時,也要控制位移量,避免由於位移過大或過小影響後續步驟。在一些實施例中,通過多次重新裝設移液管,以獲得更加準確的尖端與中心點之間的對焦距離。接著,獲得第三影像的清晰度值0.890014所對應的對焦距離為6260μm相對於該預設清晰度值1所對應的對焦距離6280μm的一對焦距離偏移量20μm,如第8圖中所示的Δz。In one embodiment, the preset sharpness value is the highest sharpness value 1 in the above mapping relationship table, and the corresponding optimal focusing distance is 6280 μm. According to the calculation of the Laplace operator, the sharpness value of the second image of the tip is 0.865822, then the focus distance is increased, the pipette is moved up by 20μm, and the third image of the tip is obtained by shooting again, and the third image is calculated by the Laplace operator The clarity value of is 0.890014. According to the characteristic that the sharpness value increases with the increase of the focusing distance and the law of Gaussian distribution, it can be known that the focusing distance between the tip and the center point when shooting the second image and the third image is less than the optimal focusing distance. For optimal focusing distance, it is determined that the sharpness value 0.890014 of the third image in the mapping relationship table is located to the left of the highest sharpness value 1, and the focusing distance corresponding to the sharpness value 0.890014 of the third image is 6260 μm. Of course, when moving the pipette, the amount of displacement should also be controlled to prevent the subsequent steps from being affected by excessive or small displacement. In some embodiments, the pipette is reinstalled multiple times to obtain a more accurate focusing distance between the tip and the center point. Next, obtain a focus distance of 6260 μm corresponding to the sharpness value of 0.890014 of the third image with a focus distance offset of 20 μm relative to the focusing distance of 6280 μm corresponding to the preset sharpness value 1, as shown in Figure 8. Δz.
進一步的,在一個實施例中,移液系統的校準方法還包括設置該影像擷取裝置的成像景深,使該對焦距離偏移量的可偵測範圍位於一預設距離範圍內,其中,擷取該第三影像的步驟中,該位移大於該預設距離範圍。採用如下公式設置影像擷取裝置的成像景深d tot: 。 Further, in one embodiment, the calibration method of the pipetting system further includes setting the imaging depth of the image capturing device so that the detectable range of the focus distance offset is within a preset distance range, wherein In the step of taking the third image, the displacement is greater than the preset distance range. Use the following formula to set the imaging depth of field d tot of the image capture device: .
其中,NA是影像擷取裝置的物鏡的數值孔徑,M是物鏡的放大倍率,NA和M都是已知量,在本實施例中採用NA為0.3,M為2.66的物鏡。λ是光波波長,通常λ=0.55μm。n是移液管與物鏡之間的介質的折射率,本實施例中介質為空氣,折射率n=1。而e是影像擷取裝置可分辨的最小距離,是一個已知量,本實施例中e=2.2μm。因此,本實施例計算得到影像擷取裝置的成像景深d tot=8.86μm。當成像景深為8.86μm時,大於8.86μm的對焦距離偏移量是可偵測的,也即本實施例中對焦距離偏移量的可偵測範圍大於8.86μm。因此,擷取第三影像的步驟中,重新裝設移液管時需使移液管的尖端與影像擷取裝置的中心點之間的對焦距離產生大於8.86μm的位移,從而使影像擷取裝置所拍攝的尖端的第二影像和第三影像之間的清晰度值的變異量能否被檢測到。 Among them, NA is the numerical aperture of the objective lens of the image capturing device, M is the magnification of the objective lens, NA and M are both known quantities. In this embodiment, an objective lens with NA being 0.3 and M being 2.66 is used. λ is the wavelength of light wave, usually λ=0.55μm. n is the refractive index of the medium between the pipette and the objective lens. In this embodiment, the medium is air, and the refractive index is n=1. And e is the minimum distance that the image capturing device can distinguish, which is a known quantity. In this embodiment, e=2.2 μm. Therefore, in this embodiment, the imaging depth of field d tot of the image capturing device is calculated as 8.86 μm. When the imaging depth of field is 8.86 μm, the focus distance offset greater than 8.86 μm is detectable, that is, the detectable range of the focus distance offset in this embodiment is greater than 8.86 μm. Therefore, in the step of capturing the third image, when reinstalling the pipette, the focus distance between the tip of the pipette and the center point of the image capturing device needs to be displaced more than 8.86μm, so that the image can be captured. Whether the variation of the sharpness value between the second image and the third image of the tip captured by the device can be detected.
進一步的,獲得第二偏移量和對焦距離偏移量後,則調整移液管的位置以補償第二偏移量和對焦距離偏移量,使移液管的尖端與校準後的中心點在X坐標軸、Y坐標軸和Z坐標軸三維方向上對準,完成移液管的校準。Further, after the second offset and the focus distance offset are obtained, the position of the pipette is adjusted to compensate for the second offset and the focus distance offset, so that the tip of the pipette is aligned with the calibrated center point Align in the three-dimensional directions of the X coordinate axis, the Y coordinate axis and the Z coordinate axis to complete the calibration of the pipette.
綜上,上述實施例中的移液系統的校準方法通過影像擷取裝置擷取校準點的影像,獲得影像擷取裝置的中心點相對於校準點於X-Y平面的第一偏移量,實現影像擷取裝置所在的移液系統的轉盤於X-Y平面的校準,使校準後的影像擷取裝置用於後續校準步驟的定位;通過校準後的影像擷取裝置至少兩次擷取移液管的尖端的影像,獲得移液管的尖端於X-Y平面的第二偏移量以及於Z坐標軸的對焦距離偏移量,實現移液管的尖端於X-Y平面和Z坐標軸的校準;從而,本案的移液系統的校準方法利用了一影像擷取裝置實現移液系統的轉盤的轉動方向的校準和移液管的三維方向的校準,以實現後續移液操作和系統運行的控制。In summary, the calibration method of the pipetting system in the above embodiment captures the image of the calibration point by the image capture device, and obtains the first offset of the center point of the image capture device with respect to the calibration point on the XY plane, so as to realize the image Calibration of the turntable of the pipetting system where the capture device is located on the XY plane, so that the calibrated image capture device is used for positioning in the subsequent calibration steps; the tip of the pipette is captured at least twice by the calibrated image capture device The second offset of the tip of the pipette on the XY plane and the offset of the focus distance on the Z coordinate axis are obtained to realize the calibration of the tip of the pipette on the XY plane and the Z coordinate axis; The method for calibrating the pipetting system utilizes an image capturing device to realize the calibration of the rotation direction of the turntable of the pipetting system and the calibration of the three-dimensional direction of the pipette, so as to realize subsequent pipetting operations and system operation control.
本案還揭露一種移液系統。第13圖示出一實施例中一種移液系統的主要模組,參照第13圖所示,本實施中移液系統1包括:一影像擷取裝置21;以及一處理器41,該處理器41被配置為通過執行多條可執行指令實現:通過該影像擷取裝置21擷取一校準點的至少一第一影像;獲得該影像擷取裝置21的一中心點相對於該第一影像中的該校準點於一平面的一第一偏移量,依據該第一偏移量調整該影像擷取裝置21的位置,使該中心點與該校準點進行校準;通過校準後的該影像擷取裝置21擷取該移液系統1的一移液管31的尖端的至少一第二影像和一第三影像,其中該第三影像擷取自該尖端與校準後的該中心點的對焦距離發生位移之後;獲得該第三影像中的該尖端相對於該第二影像中的該尖端於該平面的一第二偏移量;分別依據該位移、該第三影像相對於該第二影像的清晰度值的變異量和一預設的對焦距離與清晰度值的映射關係表,獲得該第三影像的清晰度值相對於一預設清晰度值的一對焦距離偏移量;以及,依據該第二偏移量和該對焦距離偏移量調整該移液管31的位置,使該尖端與校準後的該中心點進行校準。其中,在一些實施例中,可執行指令存儲於處理器41中,在一些實施例中,可執行指令存儲於該移液系統1的一記憶體中。可執行指令的具體執行過程和原理請參照上述移液系統的校準方法實施例的描述,此處不再展開說明。This case also discloses a pipetting system. Figure 13 shows the main modules of a pipetting system in an embodiment. With reference to Figure 13, the pipetting system 1 in this embodiment includes: an
在一個實施例中,處理器41例如是一個CPU(central processing unit,中央處理器)。處理器41分別與移液系統1的影像擷取裝置21和移液管31連接,以實現第一偏移量、第二偏移量和對焦距離偏移量的計算,並根據第一偏移量、第二偏移量和對焦距離偏移量分別控制影像擷取裝置21和移液管31進行位置調整,以實現移液系統1的校準。In one embodiment, the
綜上,上述實施例的移液系統通過影像擷取裝置擷取校準點的影像,獲得影像擷取裝置的中心點相對於校準點於平面維度的第一偏移量,實現影像擷取裝置於平面維度的校準,使校準後的影像擷取裝置用於後續校準步驟的定位;通過校準後的影像擷取裝置至少兩次擷取移液管的尖端的影像,獲得移液管的尖端於平面維度的第二偏移量以及於對焦距離維度的對焦距離偏移量,實現移液管的尖端於平面維度和對焦距離維度的校準;從而,本案利用了一影像擷取裝置實現移液系統的校準,以實現後續移液操作和系統運行的控制。In summary, the pipetting system of the above embodiment captures the image of the calibration point through the image capture device, obtains the first offset of the center point of the image capture device relative to the calibration point in the plane dimension, and realizes that the image capture device is The calibration of the plane dimension enables the calibrated image capture device to be used for the positioning of the subsequent calibration steps; the image of the tip of the pipette is captured by the calibrated image capture device at least twice to obtain the tip of the pipette on the plane The second offset of the dimension and the offset of the focus distance in the focus distance dimension realize the calibration of the tip of the pipette in the plane dimension and the focus distance dimension; thus, this case uses an image capture device to achieve the pipetting system Calibration to control subsequent pipetting operations and system operation.
綜上所述,雖然本發明已以實施例揭露如上,然其並非用以限定本發明。本發明所屬技術領域中具有通常知識者,在不脫離本發明之精神和範圍內,當可作各種之更動與潤飾。因此,本發明之保護範圍當視後附之申請專利範圍所界定者為準。In summary, although the present invention has been disclosed in the above embodiments, it is not intended to limit the present invention. Those with ordinary knowledge in the technical field to which the present invention belongs can make various changes and modifications without departing from the spirit and scope of the present invention. Therefore, the protection scope of the present invention shall be subject to those defined by the attached patent application scope.
1:移液系統 10:轉盤 11:校準點 101:移液管替換區 102:移液區 21:影像擷取裝置 211:中心點 30:移液裝置 31:移液管 300:滑軌 41:處理器 Δθ:夾角 Δx、Δy、Δz:偏移量 1: Pipetting system 10: Turntable 11: Calibration point 101: Pipette replacement area 102: Pipetting area 21: Image capture device 211: Center point 30: Pipetting device 31: Pipette 300: Slide rail 41: Processor Δθ: included angle Δx, Δy, Δz: offset
第1圖示出本案一實施例中一種移液系統的結構示意圖; 第2圖示出本案一實施例中一種移液裝置的結構示意圖; 第3圖示出本案一實施例中一種移液系統的校準方法的流程示意圖; 第4圖示出本案一實施例中一影像擷取裝置擷取一校準點的一第一影像的示意圖; 第5圖示出本案一實施例中獲得一第一偏移量的流程示意圖; 第6圖示出本案一實施例中獲得校準點的直角坐標的流程示意圖; 第7圖示出本案一實施例中影像擷取裝置擷取一移液管的尖端的一第二影像的示意圖; 第8圖示出本案一實施例中影像擷取裝置擷取移液管的尖端的一第三影像的示意圖; 第9圖示出本案一實施例中獲得一第二偏移量的流程示意圖; 第10圖示出本案一實施例中獲得第三影像中的尖端的中心的一第一直角坐標的流程示意圖; 第11圖示出本案一實施例中獲得一圖元與距離的對應關係的流程示意圖; 第12圖示出本案一實施例中獲得一對焦距離偏移量的流程示意圖;以及 第13圖示出本案一實施例中一種移液系統的模組示意圖。 Figure 1 shows a schematic structural diagram of a pipetting system in an embodiment of the present case; Figure 2 shows a schematic structural diagram of a pipetting device in an embodiment of the present case; Figure 3 shows a schematic flow chart of a method for calibrating a pipetting system in an embodiment of the present case; FIG. 4 shows a schematic diagram of an image capturing device capturing a first image of a calibration point in an embodiment of the present invention; Figure 5 shows a schematic flow chart of obtaining a first offset in an embodiment of the present case; Figure 6 shows a schematic diagram of the process of obtaining the rectangular coordinates of the calibration point in an embodiment of the present case; FIG. 7 shows a schematic diagram of the image capturing device capturing a second image of the tip of a pipette in an embodiment of the present invention; FIG. 8 shows a schematic diagram of the image capturing device capturing a third image of the tip of the pipette in an embodiment of the present invention; Figure 9 shows a schematic flow chart of obtaining a second offset in an embodiment of the present case; Figure 10 shows a schematic flow chart of obtaining a first right-angle coordinate of the center of the tip in the third image in an embodiment of the present case; Figure 11 shows a schematic flow chart of obtaining the correspondence between a graphic element and a distance in an embodiment of the present case; Figure 12 shows a schematic diagram of the flow of obtaining a focus distance offset in an embodiment of the present case; and Figure 13 shows a schematic diagram of a module of a pipetting system in an embodiment of the present case.
S10~S60:各個步驟S10~S60: Each step
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