TWI470300B - Image focusing method and auto-focusing microscopic apparatus - Google Patents
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Description
本發明係關於一種影像對焦方法與顯微裝置，特別關於一種影像對焦方法與自動對焦顯微裝置。The invention relates to an image focusing method and a microscopic device, in particular to an image focusing method and an autofocus microscopic device.
自動對焦技術已被廣泛地應用在各個領域，例如監視系統、相機、工業檢測、顯微平台等方面。透過自動對焦的方式能避免技術人員因主觀判別、疲勞等個人因素而造成對焦結果的差異。此外，在需要擷取大量對焦影像的情況下，自動對焦技術更能大幅降低所耗費的人力與時間。Autofocus technology has been widely used in various fields, such as surveillance systems, cameras, industrial inspection, microscopy, and so on. The method of autofocus can avoid the difference in focus results caused by personal factors such as subjective discrimination and fatigue. In addition, autofocus technology can significantly reduce the labor and time required to capture a large number of in-focus images.
傳統自動對焦的搜尋法大多數是以全域搜尋法進行，意即在一給定的座標軸(例如Z軸)範圍中以固定間距進行連續取像，接著計算所有影像的對焦值，再以擁有最大對焦值的影像所對應的座標值作為最終對焦位置。然而，對全域搜尋法而言，若使用低解析度的影像進行分析，則會降低焦點的精確度；若使用高解析度的影像進行分析，則會大幅增加系統負擔並降低搜尋速度。Most of the traditional autofocus search methods are based on the global search method, which means continuous image capture at a fixed pitch in a given coordinate axis (for example, Z-axis), and then calculate the focus value of all images, and then have the largest The coordinate value corresponding to the image of the focus value is used as the final focus position. However, for the global search method, if low-resolution images are used for analysis, the accuracy of the focus will be reduced; if high-resolution images are used for analysis, the system load will be greatly increased and the search speed will be reduced.
因此，如何提供一種影像對焦方法與自動對焦顯微裝置，能夠兼具搜尋焦點的速度與品質，實為當前重要課題之一。Therefore, how to provide an image focusing method and an autofocus microscopic device, which can simultaneously achieve the speed and quality of the search focus, is one of the current important topics.
有鑑於上述課題，本發明之目的為提供一種能夠兼具 搜尋焦點的速度與品質之影像對焦方法與自動對焦顯微裝置。In view of the above problems, an object of the present invention is to provide a Search for the speed and quality of the focus of the image focus method and autofocus microscopy.
為達上述目的，依據本發明之一種影像對焦方法包含：一第一對焦步驟，係使用具有一第一解析度之複數第一影像搜尋一第一焦點；以及一第二對焦步驟，使用第一焦點之焦距建立一第一搜尋範圍，第一焦點位於第一搜尋範圍內，並使用具有一第二解析度之複數第二影像，在第一搜尋範圍內搜尋一第二焦點，其中第二解析度高於第一解析度。To achieve the above object, an image focusing method according to the present invention includes: a first focusing step of searching for a first focus using a plurality of first images having a first resolution; and a second focusing step using the first The focal length of the focus establishes a first search range, the first focus is located in the first search range, and uses a second image having a second resolution to search for a second focus in the first search range, wherein the second resolution The degree is higher than the first resolution.
在一實施例中，該等第一影像係藉由每隔一第一間隔進行取像而產生，該等第二影像係藉由每隔一第二間隔進行取像而產生，第二間隔小於第一間隔。In one embodiment, the first images are generated by taking images at every first interval, and the second images are generated by taking images at every second interval, and the second interval is smaller than First interval.
在一實施例中，第一對焦步驟更包含：每隔一第三間隔進行取像以產生複數第三影像，該等第三影像具有第一解析度；使用該等第三影像搜尋一第三焦點；使用第三焦點之焦距建立一第三搜尋範圍，第三焦點位於第三搜尋範圍內；以及使用該等第一影像在第三搜尋範圍內搜尋第一焦點。In an embodiment, the first focusing step further includes: taking an image every third interval to generate a plurality of third images, the third images having a first resolution; searching for a third using the third images a focus; a third search range is established using a focal length of the third focus, the third focus is within the third search range; and the first focus is searched within the third search range using the first images.
在一實施例中，第一搜尋範圍的幅度小於第三搜尋範圍的幅度。In an embodiment, the magnitude of the first search range is less than the magnitude of the third search range.
在一實施例中，該等第一影像係藉由每隔一第一間隔進行取像而產生，第一間隔小於第三間隔。In an embodiment, the first images are generated by taking images at every first interval, and the first interval is smaller than the third interval.
在一實施例中，第二對焦步驟更包含：使用第二焦點之焦距建立一第二搜尋範圍，第二焦點位於第二搜尋範圍 內；在第二搜尋範圍內每隔一第四間隔進行取像以產生複數第四影像，該等第四影像具有第二解析度；以及使用該等第四影像在第二搜尋範圍內搜尋一第四焦點。In an embodiment, the second focusing step further comprises: establishing a second search range by using a focal length of the second focus, where the second focus is located in the second search range Taking images at every fourth interval in the second search range to generate a plurality of fourth images, the fourth images having a second resolution; and searching for a second search range using the fourth images The fourth focus.
在一實施例中，第二搜尋範圍的幅度小於第一搜尋範圍的幅度。In an embodiment, the magnitude of the second search range is less than the magnitude of the first search range.
在一實施例中，該等第二影像係藉由每隔一第二間隔進行取像而產生，第四間隔小於第二間隔。In an embodiment, the second images are generated by taking images at every second interval, and the fourth interval is smaller than the second interval.
在一實施例中，第一焦點、第二焦點、第三焦點或第四焦點之搜尋係藉由一改進拉普拉斯能量和(SML)計算方式進行。In an embodiment, the searching of the first focus, the second focus, the third focus, or the fourth focus is performed by a modified Laplacian energy and (SML) calculation.
在一實施例中，第一對焦步驟與第二對焦步驟係應用於一玻片之其中一區域，玻片之另一區域之焦點搜尋係依據第二焦點之焦距進行。In one embodiment, the first focusing step and the second focusing step are applied to one of the areas of a slide, and the focus of the other area of the slide is based on the focal length of the second focus.
在一實施例中，第一對焦步驟與第二對焦步驟係應用於一玻片之其中一區域，玻片之另一區域之焦點搜尋係依據第四焦點之焦距進行。In one embodiment, the first focusing step and the second focusing step are applied to one of the areas of one slide, and the focus search of the other area of the slide is performed according to the focal length of the fourth focus.
在一實施例中，第一對焦步驟與第二對焦步驟係應用於一玻片之其中一區域，影像對焦方法更包含：對玻片之其他複數區域進行焦點搜尋，其中該等區域之焦點搜尋係依據直線形或螺旋形之方向依序進行。In an embodiment, the first focusing step and the second focusing step are applied to one of the areas of the slide, and the image focusing method further comprises: performing focus search on other plural areas of the slide, wherein the focus search of the areas The order is sequentially performed according to the direction of the straight line or the spiral shape.
為達上述目的，依據本發明之一種自動對焦顯微裝置包含一顯微裝置、一多玻片載具、一攝像裝置、一多軸驅動平台以及一處理模組。顯微裝置具有一光學機構。多玻片載具用以承載複數玻片。攝像裝置經由光學機構取得該 等玻片之至少其中之一之一影像。多軸驅動平台係承載多玻片載具以使多玻片載具進行多軸運動。處理模組係與攝像裝置及多軸驅動平台電性連接，並截取攝像裝置之影像以及控制多軸驅動平台進行多軸運動。To achieve the above object, an autofocus microscopy apparatus according to the present invention comprises a microscopic device, a multi-slide carrier, a camera device, a multi-axis drive platform, and a processing module. The microscopy device has an optical mechanism. A multi-slide carrier is used to carry a plurality of slides. The imaging device acquires the optical device An image of at least one of the slides. The multi-axis drive platform carries a multi-slide carrier for multi-axis motion of the multi-slide carrier. The processing module is electrically connected to the camera device and the multi-axis driving platform, and intercepts the image of the camera device and controls the multi-axis driving platform for multi-axis motion.
在一實施例中，多玻片載具包含至少一卡槽以容置該等玻片。In one embodiment, the multi-slide carrier includes at least one card slot for receiving the slides.
在一實施例中，該等玻片係呈一維陣列或二維陣列設置。In one embodiment, the slides are arranged in a one-dimensional array or a two-dimensional array.
承上所述，依據本發明之影像對焦方法與自動對焦顯微裝置，其係使用較低解析度的影像粗略地搜尋一焦點，並依據此焦點建立縮小的搜尋範圍，再用較高解析度的影像在此搜尋範圍中仔細地搜尋更佳的焦點，因而在對焦精準度與系統效率間取得平衡，而能兼具搜尋焦點的速度與品質。此外，本發明之自動對焦顯微裝置之多玻片載具能同時承載多個玻片，而提升自動對焦的效能。According to the above, the image focusing method and the autofocus microscopic device according to the present invention use a lower resolution image to roughly search for a focus, and based on the focus, establish a reduced search range, and then use a higher resolution. The image is carefully searched for better focus in this search range, thus balancing the focus accuracy with the system efficiency, and the speed and quality of the search focus. In addition, the multi-slide carrier of the autofocus microscopy device of the present invention can simultaneously carry a plurality of slides to improve the performance of the autofocus.
以下將參照相關圖式，說明依本發明較佳實施例之一種影像對焦方法與自動對焦顯微裝置，其中相同的元件將以相同的參照符號加以說明。DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS Hereinafter, an image focusing method and an autofocus microscopy apparatus according to a preferred embodiment of the present invention will be described with reference to the accompanying drawings, wherein the same elements will be described with the same reference numerals.
本發明不限制影像對焦方法之應用領域，可例如應用於監視系統、相機、工業檢測、顯微平台等等。本實施例係以應用於電控顯微裝置上，並能夠有效率地調整顯微裝置之Z軸位置至準確的對焦點，並自動擷取與儲存對焦後 的顯微影像，以供後續判讀或分析之用。The present invention does not limit the field of application of the image focusing method, and can be applied, for example, to a monitoring system, a camera, an industrial inspection, a microscopic platform, and the like. The embodiment is applied to an electronically controlled microscope device, and can effectively adjust the Z-axis position of the micro-device to an accurate focus point, and automatically capture and store the focus after focusing. Microscopic images for subsequent interpretation or analysis.
圖1為本發明較佳實施例之一種影像對焦方法的流程圖，圖2為本發明第一實施例之一種影像對焦方法的解說示意圖。如圖1及圖2所示，本實施例之影像對焦方法包含一第一對焦步驟，係使用具有一第一解析度之複數第一影像搜尋一第一焦點(步驟S01)。第一解析度為較低的解析度，可例如但不限於320×240。該等第一影像可藉由每隔一第一間隔進行取像而產生。在本實施例中，第一影像係藉由一攝像器沿一Z軸移動，例如從任一點(例如-85μm)向上每隔第一間隔取像一次而產生，第一間隔於此為6μm。於此，第一影像係以從-85μm至-115μm取像而得到6張為例。第一焦點之搜尋可藉由一改進拉普拉斯能量和(sum-modified Laplacian,SML)計算方式進行而得到較佳效果；但本發明不限制使用其他計算方式。焦點可藉由影像銳利度之估算來達成。在上述6張第一影像中，具有最大之SML值的即是有最佳對焦點的影像，藉此可找到第一焦點。於此，第一焦點例如是-109μm。需注意者，焦點的座標值會依據座標系的改變而變化，於此僅為舉例說明。1 is a flow chart of an image focusing method according to a preferred embodiment of the present invention, and FIG. 2 is a schematic diagram of an image focusing method according to a first embodiment of the present invention. As shown in FIG. 1 and FIG. 2, the image focusing method of this embodiment includes a first focusing step of searching for a first focus using a plurality of first images having a first resolution (step S01). The first resolution is a lower resolution, which may be, for example but not limited to, 320×240. The first images can be generated by taking images at every first interval. In the present embodiment, the first image is generated by a camera moving along a Z axis, for example, from any point (for example, -85 μm) upward at every first interval, and the first interval is 6 μm. Here, the first image is obtained by taking six images from -85 μm to -115 μm. The search for the first focus can be performed by a modified sum-modified Laplacian (SML) calculation method; however, the present invention does not limit the use of other calculation methods. The focus can be achieved by estimating the image sharpness. Among the above six first images, the image having the largest SML value is the image having the best focus point, whereby the first focus can be found. Here, the first focus is, for example, -109 μm. It should be noted that the coordinate value of the focus will change according to the change of the coordinate system, which is only an example.
本實施例之影像對焦方法更包含一第二對焦步驟，其係使用第一焦點之焦距建立一第一搜尋範圍，第一焦點位於第一搜尋範圍內，並使用具有一第二解析度之複數第二影像，在第一搜尋範圍內搜尋一第二焦點，其中第二解析度高於第一解析度(步驟S02)。The image focusing method of this embodiment further includes a second focusing step of establishing a first search range by using a focal length of the first focus, the first focus is within the first search range, and using a plurality of second resolutions The second image searches for a second focus within the first search range, wherein the second resolution is higher than the first resolution (step S02).
在得到第一焦點之後，即利用第一焦點之焦距建立一第一搜尋範圍，於此第一搜尋範圍為-105μm~-113μm，且第一焦點-109μm係位於第一搜尋範圍內。在本實施例中，第一搜尋範圍係以第一焦點為中心，以±4μm為間距而建立。而在其他實施例中，搜尋範圍可由其他演算法建立，例如不對稱方式、權重方式等等。在建立第一搜尋範圍之後，即使用具有第二解析度之第二影像，在第一搜尋範圍內搜尋一第二焦點。第二解析度高於第一解析度，例如但不限於1280×960。理論上，取像解析度越高，影像所含的資訊會越豐富，用它來估算對焦值會越精準。After the first focus is obtained, a first search range is established by using the focal length of the first focus, where the first search range is -105 μm - 113 μm, and the first focus - 109 μm is located within the first search range. In this embodiment, the first search range is centered on the first focus and is established with a pitch of ±4 μm. In other embodiments, the search range may be established by other algorithms, such as an asymmetric mode, a weighting mode, and the like. After the first search range is established, a second image with a second resolution is used to search for a second focus within the first search range. The second resolution is higher than the first resolution, such as but not limited to 1280×960. In theory, the higher the image resolution, the richer the information contained in the image, and the more accurate it is to estimate the focus value.
該等第二影像可藉由每隔一第二間隔進行取像而產生，且第二間隔小於第一間隔。於此，第二影像係藉由一攝像器沿Z軸移動，例如從第一搜尋範圍的起點(如-105μm)向上每隔第二間隔取像一次而產生，第二間隔於此為4μm，其小於第一間隔6μm。於此，第二影像係以3張為例。第二焦點之搜尋可藉由一改進拉普拉斯能量和(SML)計算方式進行。在上述3張第一影像中，具有最大之SML值的即是有最佳對焦點的影像，藉此可找到第二焦點。於此，第二焦點例如是-113μm。據此，本實施例係使用較低解析度的影像粗略地搜尋一焦點，再用較高解析度的影像在小範圍中仔細地搜尋最佳的焦點，因而在對焦精準度與系統效率間取得平衡，而能兼具搜尋焦點的速度與品質。The second images may be generated by taking images at every second interval, and the second interval is smaller than the first interval. Here, the second image is generated by a camera moving along the Z axis, for example, from the starting point of the first search range (eg, -105 μm) up to the second interval, and the second interval is 4 μm. It is smaller than the first interval by 6 μm. Here, the second image is taken as an example of three sheets. The search for the second focus can be performed by a modified Laplacian energy and (SML) calculation. Among the above three first images, the image having the largest SML value is the image having the best focus point, whereby the second focus can be found. Here, the second focus is, for example, -113 μm. Accordingly, the present embodiment uses a lower resolution image to roughly search for a focus, and then uses a higher resolution image to carefully search for the best focus in a small range, thereby achieving between focus accuracy and system efficiency. Balance, but the speed and quality of the search focus.
圖3為本發明第二實施例之一種影像對焦方法的解說 示意圖。如圖3所示，第一對焦步驟更包含：每隔一第三間隔進行取像以產生複數第三影像，該等第三影像具有第一解析度；以及使用該等第三影像搜尋一第三焦點。在本實施例中，第三影像係藉由一攝像器沿Z軸移動，例如從任一點(例如100μm)向上每隔一第三間隔取像一次而產生，第三間隔於此為20μm，且第三間隔係大於第二間隔與第一間隔。於此，第三影像係從100μm至-260μm取像而得到19張為例，且第三影像與第一影像之解析度相同，皆為320×240。而在其他實施例中，第三影像與第一影像之解析度可不相同。第三焦點之搜尋可藉由一改進拉普拉斯能量和(SML)計算方式進行。在上述19張第一影像中，具有最大之SML值的即是有最佳對焦點的影像，藉此可找到第三焦點。於此，第三焦點例如是-100μm。3 is an illustration of an image focusing method according to a second embodiment of the present invention; schematic diagram. As shown in FIG. 3, the first focusing step further includes: taking an image at every third interval to generate a plurality of third images, the third images having a first resolution; and searching for the third image using the third image Three focus. In this embodiment, the third image is generated by a camera moving along the Z axis, for example, taking images from every point (for example, 100 μm) up to every third interval, and the third interval is 20 μm, and The third spacing is greater than the second spacing and the first spacing. Here, the third image is taken from 100 μm to -260 μm to obtain 19 images, and the resolution of the third image and the first image are both 320×240. In other embodiments, the resolution of the third image and the first image may be different. The search for the third focus can be performed by a modified Laplacian energy and (SML) calculation. Among the above 19 first images, the image having the largest SML value is the image having the best focus point, whereby the third focus can be found. Here, the third focus is, for example, -100 μm.
然後，第一對焦步驟更包含：使用第三焦點之焦距建立一第三搜尋範圍，第三焦點位於第三搜尋範圍內；以及使用該等第一影像在第三搜尋範圍內搜尋第一焦點。在得到第三焦點之後，即利用第三焦點之焦距建立一第三搜尋範圍，於此第三搜尋範圍為-85μm~-115μm，且第三搜尋範圍的幅度(margin)大於第一搜尋範圍的幅度。於此，第三搜尋範圍的幅度為30μm(| -85-(-115)|)，第一搜尋範圍的幅度為8μm(| -105-(-113)|)。並且第三焦點-100μm係位於第三搜尋範圍內。在本實施例中，第三搜尋範圍係以第三焦點為中心，以±15μm為間距而建立。在建立第三搜尋範圍之後，即使用第一影像在第三搜尋範圍內 搜尋第一焦點。由於第一焦點之搜尋已在第一實施例詳述，故於此不再贅述。Then, the first focusing step further includes: establishing a third search range by using a focal length of the third focus, the third focus is located in the third search range; and searching for the first focus in the third search range by using the first images. After the third focus is obtained, a third search range is established by using the focal length of the third focus, where the third search range is -85 μm to -115 μm, and the third search range has a larger margin than the first search range. Amplitude. Here, the amplitude of the third search range is 30 μm (| -85-(-115)|), and the amplitude of the first search range is 8 μm (| -105-(-113)|). And the third focus - 100 μm is located in the third search range. In this embodiment, the third search range is centered on the third focus and is established with a pitch of ±15 μm. After the third search range is established, the first image is used in the third search range. Search for the first focus. Since the search for the first focus has been described in detail in the first embodiment, it will not be described herein.
圖4為本發明第三實施例之一種影像對焦方法的解說示意圖。如圖4所示，第二對焦步驟更包含：使用第二焦點之焦距建立一第二搜尋範圍，第二焦點位於第二搜尋範圍內。在上述第一實施例中已取得第二焦點-113μm，然後，可使用第二焦點之焦距建立一第二搜尋範圍。於此第二搜尋範圍為-111μm~-115μm，且第二焦點-113μm係位於第二搜尋範圍內。在本實施例中，第二搜尋範圍係以第二焦點為中心，以±2μm為間距而建立。FIG. 4 is a schematic diagram of an image focusing method according to a third embodiment of the present invention. As shown in FIG. 4, the second focusing step further includes: establishing a second search range by using a focal length of the second focus, the second focus being located in the second search range. In the first embodiment described above, the second focus is -113 μm, and then a second search range can be established using the focal length of the second focus. The second search range is -111μm~-115μm, and the second focus-113μm is located in the second search range. In this embodiment, the second search range is centered on the second focus and is established with a pitch of ±2 μm.
然後，第二對焦步驟更包含：在第二搜尋範圍內每隔一第四間隔進行取像以產生複數第四影像，該等第四影像具有該第二解析度。在建立第二搜尋範圍之後，即在第二搜尋範圍內每隔一第四間隔進行取像以產生複數第四影像。於此，第二影像係藉由一攝像器沿Z軸移動，例如從第二搜尋範圍的起點(如-111μm)向上每隔第四間隔取像一次而產生，第四間隔於此為2μm，且第四間隔小於第一間隔(6μm)、第二間隔(4μm)與第三間隔(20μm)。於此，第四影像係以從-111μm至-115μm取像而得到3張為例。並且第四影像與第二影像之解析度相同，皆為1280×960。而在其他實施例中，第四影像與第二影像之解析度可不相同，例如高於第二影像之解析度。Then, the second focusing step further includes: taking an image at every fourth interval in the second search range to generate a plurality of fourth images, the fourth images having the second resolution. After the second search range is established, the image is taken at every fourth interval in the second search range to generate a plurality of fourth images. In this case, the second image is generated by a camera moving along the Z axis, for example, from the starting point of the second search range (eg, -111 μm) to the fourth interval, and the fourth interval is 2 μm. And the fourth interval is smaller than the first interval (6 μm), the second interval (4 μm), and the third interval (20 μm). Here, the fourth image is obtained by taking three images from -111 μm to -115 μm. And the resolution of the fourth image and the second image are the same, both being 1280×960. In other embodiments, the resolution of the fourth image and the second image may be different, for example, higher than the resolution of the second image.
然後，第二對焦步驟更包含：使用該等第四影像在第二搜尋範圍內搜尋一第四焦點。第四焦點之搜尋可藉由一 改進拉普拉斯能量和(SML)計算方式進行。在上述3張第四影像中，具有最大之SML值的即是有最佳對焦點的影像，藉此可找到第四焦點。於此，第四焦點例如是-111μm。Then, the second focusing step further includes: searching for a fourth focus in the second search range by using the fourth images. The fourth focus can be searched by one Improved Laplacian energy and (SML) calculations are performed. Among the above three fourth images, the image having the largest SML value is the image having the best focus point, whereby the fourth focus can be found. Here, the fourth focus is, for example, -111 μm.
於此，第二搜尋範圍的幅度(margin)小於第一搜尋範圍的幅度。第二搜尋範圍為4μm(| -111-(-115)|)，第一搜尋範圍的幅度為8μm(| -105-(-113)|)。藉此可見本發明之自動對焦方法之焦點搜尋可將搜尋範圍逐漸縮小，並找到最佳的對焦點。Here, the margin of the second search range is smaller than the amplitude of the first search range. The second search range is 4 μm (| -111-(-115)|), and the amplitude of the first search range is 8 μm (| -105-(-113)|). It can be seen that the focus search of the autofocus method of the present invention can gradually narrow the search range and find the best focus point.
在本發明中，第一對焦步驟與第二對焦步驟可個別不斷延伸，而產生更多的搜尋範圍以搜尋更精確的焦點。甚至亦可使用三種以上之解析度的影像來進行影像對焦方法，而進一步提升焦點搜尋的精確度及效能。In the present invention, the first focusing step and the second focusing step can be individually extended to generate more search ranges to search for more precise focus. It is even possible to use more than three resolution images for image focusing, which further improves the accuracy and performance of focus search.
本發明之影像對焦方法可應用於一玻片，玻片上可載有一待檢測體，例如是病菌體。在上述第一實施例中(請參照圖2)，第一對焦步驟與第二對焦步驟係應用於一玻片之其中一區域，而玻片之另一區域之焦點搜尋可依據第二焦點之焦距進行，如此可避免重新搜尋，因而提升搜尋效率。而在上述第三實施例中(請參照圖4)，第一對焦步驟與第二對焦步驟係應用於一玻片之其中一區域，而玻片之另一區域之焦點搜尋可依據第四焦點之焦距進行，如此同樣可提升搜尋效率。總而言之，玻片上另一區域之焦點搜尋可依據之前找到的焦點來進行，例如依據第一焦點至第四焦點之任一來進行。The image focusing method of the present invention can be applied to a slide, which can carry a sample to be detected, such as a pathogen. In the first embodiment described above (please refer to FIG. 2), the first focusing step and the second focusing step are applied to one of the areas of one of the slides, and the focus of the other area of the slide is searchable according to the second focus. The focal length is carried out, so that re-searching can be avoided, thereby improving search efficiency. In the above third embodiment (please refer to FIG. 4), the first focusing step and the second focusing step are applied to one of the areas of one slide, and the focus of the other area of the slide can be based on the fourth focus. The focal length is carried out, which also improves the search efficiency. In summary, the focus search for another area on the slide can be made based on the previously found focus, for example, depending on either the first focus or the fourth focus.
另外，當影像對焦方法應用於一玻片的多個區域時，該等區域之焦點搜尋可依據不同的路線來進行，例如該等區域可以直線形或螺旋形之方向依序進行影像對焦方法。並且其中一個區域所得到的焦點可作為其他區域之焦點搜尋的依據，例如其他區域的焦點搜尋可在第一區域所得到的焦點所建立的搜尋範圍內搜尋，進而提升搜尋效能及精確度。圖5為一玻片G之多個區域A進行影像對焦方法的路徑示意圖，其中係以直線形為例。In addition, when the image focusing method is applied to a plurality of regions of a slide, the focus search of the regions may be performed according to different routes, for example, the regions may perform image focusing methods in a straight line or a spiral direction. And the focus obtained by one of the regions can be used as the basis for the focus search of other regions. For example, the focus search in other regions can search within the search range established by the focus obtained in the first region, thereby improving search performance and accuracy. FIG. 5 is a schematic diagram of a path for performing image focusing on a plurality of regions A of a slide glass G, wherein a straight line is taken as an example.
圖6為本發明較佳實施例之一種自動對焦顯微裝置1的示意圖。請參照圖6，自動對焦顯微裝置1包含一顯微裝置11、一多玻片載具12、一攝像裝置13、一多軸驅動平台14以及一處理模組15。Figure 6 is a schematic illustration of an autofocus microscopy apparatus 1 in accordance with a preferred embodiment of the present invention. Referring to FIG. 6 , the autofocus microscopy apparatus 1 includes a microscopy device 11 , a multi-slide carrier 12 , an imaging device 13 , a multi-axis drive platform 14 , and a processing module 15 .
本實施例不限制顯微裝置11之類別、尺寸或結構等等。於此，顯微裝置11具有一光學機構，光學機構的設計可依實際需要而調整。光學機構例如包含至少一目鏡與至少一物鏡以觀察至少一玻片上之待檢測體。此外，光學機構可更包含多個透鏡以調整光線。另外，在其他實施例中，目鏡之設置或可省略，而由攝像裝置13直接進行攝像。This embodiment does not limit the type, size or structure of the microscopic device 11, and the like. Here, the micro-device 11 has an optical mechanism, and the design of the optical mechanism can be adjusted according to actual needs. The optical mechanism comprises, for example, at least one eyepiece and at least one objective lens to view the object to be detected on at least one of the slides. In addition, the optical mechanism may further include a plurality of lenses to adjust the light. In addition, in other embodiments, the setting of the eyepiece may be omitted, and the imaging device 13 directly performs imaging.
多玻片載具12用以承載複數玻片G。習知的玻片載具僅能承載單一玻片，而本發明之多玻片載具經過改良而能同時承載多個玻片，進而提升檢測效能。圖7為本發明較佳實施例之一種多玻片載具12承載玻片G的示意圖，圖8為圖7之多玻片載具12與玻片G分離的示意圖。如圖7 與圖8所示，多玻片載具12包含至少一卡槽121以容置該等玻片G，於此係以包含二卡槽121為例，並可承載六玻片。該等玻片G係呈一維陣列或二維陣列設置，於此係以呈二維陣列設置為例。The multi-slide carrier 12 is used to carry a plurality of slides G. The conventional slide carrier can only carry a single slide, and the multi-slide carrier of the present invention can be modified to carry multiple slides at the same time, thereby improving the detection efficiency. FIG. 7 is a schematic diagram of a multi-slide carrier 12 carrying a slide G according to a preferred embodiment of the present invention, and FIG. 8 is a schematic view of the multi-slide carrier 12 of FIG. 7 separated from the slide G. Figure 7 As shown in FIG. 8, the multi-slide carrier 12 includes at least one card slot 121 for accommodating the slides G. Here, the two-card slot 121 is included as an example, and can carry six slides. The slides G are arranged in a one-dimensional array or a two-dimensional array, and are set as a two-dimensional array.
攝像裝置13經由光學機構取得該等玻片G之至少其中之一之複數影像。於此，光學機構之物鏡係對準目標玻片，使得攝像裝置13經由光學機構取得該玻片之影像。攝像裝置13例如包含電荷耦合元件(CCD)、互補式金氧半感測元件(CMOS)或其他感光元件以感應光線而形成影像。The imaging device 13 acquires a plurality of images of at least one of the slides G via an optical mechanism. Here, the objective lens of the optical mechanism is aligned with the target slide, so that the imaging device 13 acquires the image of the slide via the optical mechanism. The imaging device 13 includes, for example, a charge coupled device (CCD), a complementary MOS semi-sense element (CMOS), or other photosensitive element to induce light to form an image.
多軸驅動平台14係承載多玻片載具12以使多玻片載具12進行多軸運動。於此，多軸運動包含X-Y軸之平面運動以及Z軸之垂直運動。The multi-axis drive platform 14 carries a multi-slide carrier 12 for multi-axis motion of the multi-slide carrier 12. Here, the multi-axis motion includes the plane motion of the X-Y axis and the vertical motion of the Z axis.
處理模組15係與攝像裝置13及多軸驅動平台14電性連接，並截取攝像裝置13之影像以及控制多軸驅動平台14進行多軸運動。處理模組15例如由一電腦實現。The processing module 15 is electrically connected to the imaging device 13 and the multi-axis driving platform 14, and intercepts the image of the imaging device 13 and controls the multi-axis driving platform 14 to perform multi-axis motion. The processing module 15 is implemented, for example, by a computer.
在本實施例中，影像對焦方法可應用於自動對焦顯微裝置1。其中，處理模組15可建立一搜尋範圍，並依據該搜尋範圍控制多軸驅動平台14垂直移動，同時控制攝像裝置13進行取像，並儲存攝像裝置13所取得之影像，例如第一影像。然後，處理模組15係利用一計算方法，例如改進拉普拉斯能量和(SML)來計算而得到焦點，例如第一焦點。接著，處理模組15使用得到之焦點來建立另一搜尋範圍，例如第一搜尋範圍。然後處理模組15控制 攝像裝置13以改變其影像解析度，例如調整為第二解析度。然後處理模組15控制多軸驅動平台14依據第一搜尋範圍垂直移動，同時控制攝像裝置13進行取像，並儲存攝像裝置13所取得之影像，例如第二影像。接著利用一計算方法計算而得到焦點，例如第二焦點。藉由上述方式可例如得到第一焦點至第四焦點。In the present embodiment, the image focusing method can be applied to the autofocus microscopy apparatus 1. The processing module 15 can establish a search range, and control the vertical movement of the multi-axis driving platform 14 according to the search range, and simultaneously control the imaging device 13 to perform image capturing, and store an image obtained by the camera device 13, such as a first image. The processing module 15 then calculates the focus using a computational method, such as improved Laplacian energy and (SML), such as the first focus. Next, the processing module 15 uses the resulting focus to establish another search range, such as the first search range. Then the processing module 15 controls The imaging device 13 changes its image resolution, for example, to the second resolution. Then, the processing module 15 controls the multi-axis driving platform 14 to vertically move according to the first search range, and simultaneously controls the imaging device 13 to perform image capturing, and stores an image obtained by the camera device 13, such as a second image. Then use a calculation method to calculate the focus, such as the second focus. The first focus to the fourth focus can be obtained, for example, by the above method.
此外，處理模組15得到一玻片上之一區域之最佳焦點之後，可控制多軸驅動平台14進行平面移動，以使該玻片之另一區域對準光學機構之物鏡，以進行另一區域之焦點搜尋。並且另一區域之焦點搜尋可依據上一區域之最佳焦點來進行，而縮短搜尋時間並提升搜尋效能。In addition, after the processing module 15 obtains the best focus of a region on a slide, the multi-axis driving platform 14 can be controlled to move in a plane so that another region of the slide is aligned with the objective lens of the optical mechanism to perform another Focus search for the area. And the focus search of another area can be based on the best focus of the previous area, which shortens the search time and improves the search performance.
此外，由於本實施例之自動對焦顯微裝置1之玻片載具為「多」玻片載具，因此能不中斷地檢測一批次的玻片，而大幅提升檢測效能，並且另一玻片的焦點搜尋亦可利用已進行焦點搜尋之玻片之其中一焦點來進行。In addition, since the slide carrier of the autofocus microscopy apparatus 1 of the present embodiment is a "multiple" slide carrier, it is possible to detect a batch of slides without interruption, and greatly improve the detection efficiency, and another glass The focus search of the slice can also be performed using one of the focal points of the slide that has been searched for focus.
綜上所述，依據本發明之影像對焦方法與自動對焦顯微裝置，其係使用較低解析度的影像粗略地搜尋一焦點，並依據此焦點建立縮小的搜尋範圍，再用較高解析度的影像在此搜尋範圍中仔細地搜尋更佳的焦點，因而在對焦精準度與系統效率間取得平衡，而能兼具搜尋焦點的速度與品質。此外，本發明之自動對焦顯微裝置之多玻片載具能同時承載多個玻片，而提升自動對焦的效能。In summary, the image focusing method and the autofocus microscopic device according to the present invention use a lower resolution image to roughly search for a focus, and based on the focus, establish a reduced search range, and then use a higher resolution. The image is carefully searched for better focus in this search range, thus balancing the focus accuracy with the system efficiency, and the speed and quality of the search focus. In addition, the multi-slide carrier of the autofocus microscopy device of the present invention can simultaneously carry a plurality of slides to improve the performance of the autofocus.
以上所述僅為舉例性，而非為限制性者。任何未脫離本發明之精神與範疇，而對其進行之等效修改或變更，均 應包含於後附之申請專利範圍中。The above is intended to be illustrative only and not limiting. Any equivalent modifications or variations to the spirit and scope of the invention are It should be included in the scope of the patent application attached.
1‧‧‧自動對焦顯微裝置1‧‧‧Autofocus microscopy
11‧‧‧顯微裝置11‧‧‧Microscopic device
12‧‧‧多玻片載具12‧‧‧Multi-slide carrier
121‧‧‧卡槽121‧‧‧ card slot
13‧‧‧攝像裝置13‧‧‧ camera
14‧‧‧多軸驅動平台14‧‧‧Multi-axis drive platform
15‧‧‧處理模組15‧‧‧Processing module
A‧‧‧區域A‧‧‧ area
G‧‧‧玻片G‧‧‧ slides
S01~S02‧‧‧影像對焦方法的步驟S01~S02‧‧‧ steps of image focusing method
圖1為本發明較佳實施例之一種影像對焦方法的流程圖；圖2為本發明第一實施例之一種影像對焦方法的解說示意圖；圖3為本發明第二實施例之一種影像對焦方法的解說示意圖；圖4為本發明第三實施例之一種影像對焦方法的解說示意圖；圖5為一玻片之多個區域進行影像對焦方法的路徑示意圖，其中係以直線形為例；圖6為本發明較佳實施例之一種自動對焦顯微裝置的示意圖；圖7為本發明較佳實施例之一種多玻片載具承載玻片的示意圖；以及圖8為圖7之多玻片載具與玻片分離的示意圖。1 is a flowchart of an image focusing method according to a preferred embodiment of the present invention; FIG. 2 is a schematic diagram of an image focusing method according to a first embodiment of the present invention; FIG. 3 is a schematic image focusing method according to a second embodiment of the present invention; FIG. 4 is a schematic diagram of an image focusing method according to a third embodiment of the present invention; FIG. 5 is a schematic diagram of a path for performing image focusing on a plurality of regions of a slide, wherein a straight line is taken as an example; A schematic diagram of an autofocus microscopy apparatus according to a preferred embodiment of the present invention; FIG. 7 is a schematic view of a multi-slide carrier carrier slide according to a preferred embodiment of the present invention; and FIG. 8 is a multi-slice slide of FIG. A schematic diagram separated from the slide.
S01~S02‧‧‧影像對焦方法的步驟S01~S02‧‧‧ steps of image focusing method
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| TW200611065A (en) * | 2004-09-23 | 2006-04-01 | Asia Optical Co Inc | Auto focus methods and auto focus devices for electronic cameras using the same |
| TW200620990A (en) * | 2004-12-10 | 2006-06-16 | Hon Hai Prec Ind Co Ltd | System and method for focusing images automatically |
| JP2008079123A (en) * | 2006-09-22 | 2008-04-03 | Olympus Corp | Imaging apparatus and focus control program |
| TW201044005A (en) * | 2009-06-05 | 2010-12-16 | Univ Chung Yuan Christian | Method for auto focus searching of optical microscope |
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