CN105866937B - A method of it being converted to visible light using laser and determines micro objective focus - Google Patents
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- 238000000034 method Methods 0.000 title claims abstract description 40
- 239000013078 crystal Substances 0.000 claims abstract description 100
- 238000006243 chemical reaction Methods 0.000 claims abstract description 37
- 238000004020 luminiscence type Methods 0.000 claims abstract description 11
- QWVYNEUUYROOSZ-UHFFFAOYSA-N trioxido(oxo)vanadium;yttrium(3+) Chemical compound [Y+3].[O-][V]([O-])([O-])=O QWVYNEUUYROOSZ-UHFFFAOYSA-N 0.000 claims description 30
- WUPHOULIZUERAE-UHFFFAOYSA-N 3-(oxolan-2-yl)propanoic acid Chemical compound OC(=O)CCC1CCCO1 WUPHOULIZUERAE-UHFFFAOYSA-N 0.000 claims description 18
- 229910052980 cadmium sulfide Inorganic materials 0.000 claims description 18
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Abstract
本发明涉及确定显微镜物镜焦点的技术领域，尤其涉及一种应用激光转换为可见光确定显微镜物镜焦点的方法，该方法是以不可见激光作为显微镜的光源，将转换发光晶体置于显微镜的载物台上，并对准显微镜的通光孔中央，改变显微镜的物镜与转换发光晶体之间的距离，使不可见激光照射在转换发光晶体上，在转换发光晶体内聚焦出现有颜色的可见光点，该可见光点即为显微镜物镜焦点，该可见光点所在位置即为显微镜物镜焦点位置。本发明可以快速、直观、准确的确定显微镜物镜焦点的空间位置，以便后期将被测物体调节至显微镜物镜焦点所在焦平面位置，从而简易调节出显微镜中被测物体的清晰图像。
The invention relates to the technical field of determining the focal point of the microscope objective lens, in particular to a method for determining the focus of the microscope objective lens by converting laser light into visible light. The method uses invisible laser light as the light source of the microscope, and places the converted luminescent crystal on the stage of the microscope and aim at the center of the light hole of the microscope, change the distance between the objective lens of the microscope and the conversion luminescent crystal, so that the invisible laser light is irradiated on the conversion luminescence crystal, and a colored visible light spot appears in the conversion luminescence crystal. The visible light point is the focus of the microscope objective lens, and the position of the visible light point is the focus position of the microscope objective lens. The invention can quickly, intuitively and accurately determine the spatial position of the focus of the objective lens of the microscope, so that the object to be measured can be adjusted to the focal plane position where the focus of the objective lens of the microscope is located in the later stage, thereby easily adjusting a clear image of the object to be measured in the microscope.
Description
技术领域technical field
本发明涉及确定显微镜物镜焦点的技术领域，尤其涉及一种应用激光转换为可见光确定显微镜物镜焦点的方法。The invention relates to the technical field of determining the focus of a microscope objective lens, in particular to a method for determining the focus of a microscope objective lens by converting laser light into visible light.
背景技术Background technique
光学显微镜是利用光学原理，呈现人眼所不能分辨的微细结构的光学仪器。基于基本的几何光学原理，如果能较为便捷的确定光学显微镜在不同倍数物镜下的焦点位置，则实际操作过程中便能较为迅速地调节出被测物体清晰的图像。现有技术中，常见显微镜的自动聚焦系统通常是以计算机为控制核心，采用步进电机等作为运动控制部件，选用CCD或者CMOS摄像头结合软件算法来进行聚焦分析和运动控制；该种方法理论计算过程繁琐，设备成本高，使用场合有限，无法应用于平时实验过程。An optical microscope is an optical instrument that uses optical principles to present fine structures that cannot be resolved by the human eye. Based on the basic principles of geometric optics, if the focus position of the optical microscope under different magnification objective lenses can be determined more conveniently, then a clear image of the measured object can be adjusted more quickly during the actual operation. In the prior art, the automatic focusing system of a common microscope usually uses a computer as the control core, uses a stepping motor as a motion control component, and uses a CCD or CMOS camera combined with a software algorithm to perform focus analysis and motion control; the theoretical calculation of this method The process is cumbersome, the equipment cost is high, and the use occasions are limited, so it cannot be applied to the usual experimental process.
发明内容Contents of the invention
本发明的目的在于针对现有技术的不足，提供一种设计合理，操作简便，直观，高效，经济的应用激光转换为可见光确定显微镜物镜焦点的方法。The object of the present invention is to aim at the deficiencies of the prior art, to provide a reasonable design, easy to operate, intuitive, efficient and economical method of converting laser light into visible light to determine the focal point of the microscope objective lens.
为实现上述目的，本发明采用以下技术方案：To achieve the above object, the present invention adopts the following technical solutions:
一种应用激光转换为可见光确定显微镜物镜焦点的方法，该方法是以不可见激光作为显微镜的光源，将转换发光晶体置于显微镜的载物台上，并对准显微镜的通光孔中央，改变显微镜的物镜与转换发光晶体之间的距离，使不可见激光照射在转换发光晶体上，在转换发光晶体内聚焦出现有颜色的可见光点，该可见光点即为显微镜物镜焦点，该可见光点所在位置即为显微镜物镜焦点位置。A method of converting laser light into visible light to determine the focal point of the microscope objective lens. The method uses invisible laser light as the light source of the microscope, places the converted luminescent crystal on the stage of the microscope, and aligns it with the center of the aperture of the microscope. The distance between the objective lens of the microscope and the converted luminescent crystal is such that the invisible laser light is irradiated on the converted luminescent crystal, and a colored visible light spot appears after focusing in the converted luminescent crystal. The visible light spot is the focus of the microscope objective lens. The position of the visible light spot is is the focal point of the microscope objective lens.
作为优选，所述转换发光晶体为上转换发光晶体或者下转换发光晶体。Preferably, the conversion light-emitting crystal is an up-conversion light-emitting crystal or a down-conversion light-emitting crystal.
作为优选，所述的上转换发光晶体为掺铒钒酸钇晶体，所述的不可见激光的波长为980nm，所述的可见光点为绿色。本发明采用的掺铒钒酸钇晶体可以把低能光子转换成高能光子，发光效率高，显示光路清晰，具有良好的机械性质和物理性质，在诸多方面所显示的优良性质，尤其适于作为激光晶体材料；当波长为980nm的肉眼不可见激光照射在掺铒钒酸钇晶体上时，可以看到掺铒钒酸钇晶体内部显示出波长为553nm的肉眼可见绿色光子，通过改变显微镜的物镜与掺铒钒酸钇晶体之间的距离，使得掺铒钒酸钇晶体内部出现绿色的可见光点，该绿色可见光点即为显微镜物镜焦点，然后通过测量显微镜的物镜与载物台之间的距离A，或者测量显微镜的光源与载物台之间的距离B，从而确定了显微镜的物镜焦点在空间上的位置。Preferably, the up-conversion luminescent crystal is an erbium-doped yttrium vanadate crystal, the wavelength of the invisible laser is 980nm, and the visible light point is green. The erbium-doped yttrium vanadate crystal used in the present invention can convert low-energy photons into high-energy photons, has high luminous efficiency, shows a clear optical path, and has good mechanical and physical properties. The excellent properties shown in many aspects are especially suitable as laser Crystal material; when the invisible laser with a wavelength of 980nm is irradiated on the erbium-doped yttrium vanadate crystal, it can be seen that the inside of the erbium-doped yttrium vanadate crystal shows green photons with a wavelength of 553nm visible to the naked eye. By changing the objective lens of the microscope and The distance between the erbium-doped yttrium vanadate crystals makes a green visible light point appear inside the erbium-doped yttrium vanadate crystal, and the green visible light point is the focus of the microscope objective lens, and then by measuring the distance A between the objective lens of the microscope and the stage , or measure the distance B between the light source of the microscope and the stage, so as to determine the position of the focal point of the objective lens of the microscope in space.
进一步，所述的掺铒钒酸钇晶体采用提拉法制得，掺铒钒酸钇晶体的的掺铒浓度为0.2-1.2mol%。本发明采用掺铒浓度为0.2-1.2mol%的掺铒钒酸钇晶体使得绿光具有较长的荧光寿命，而且分辨率好，成本低。Further, the erbium-doped yttrium vanadate crystal is prepared by a pulling method, and the erbium-doped concentration of the erbium-doped yttrium vanadate crystal is 0.2-1.2 mol%. The invention adopts the erbium-doped yttrium vanadate crystal with the erbium-doped concentration of 0.2-1.2 mol%, so that the green light has a long fluorescence lifetime, good resolution and low cost.
作为优选，所述的下转换发光晶体为硫化镉晶体，所述的可见激光的波长为325nm，所述的可见光点为蓝绿色。本发明采用的硫化镉晶体可以把高能光子转换成低能光子，当波长为325nm的肉眼不可见激光照射在硫化镉晶体上时，可以看到硫化镉晶体内部显示出肉眼可见的蓝绿色光子，通过改变显微镜的物镜与掺铒钒酸钇晶体之间的距离，使得硫化镉晶体内部出现蓝绿色的可见光点，该蓝绿色可见光点即为显微镜的物镜焦点，然后通过测量显微镜的物镜与载物台之间的距离A，或者测量显微镜的光源与载物台之间的距离B，从而确定了显微镜的物镜焦点在空间上的位置。Preferably, the down-conversion luminescent crystal is cadmium sulfide crystal, the wavelength of the visible laser is 325nm, and the visible light spot is blue-green. The cadmium sulfide crystal adopted in the present invention can convert high-energy photons into low-energy photons. When an invisible laser with a wavelength of 325nm is irradiated on the cadmium sulfide crystal, it can be seen that the inside of the cadmium sulfide crystal shows blue-green photons visible to the naked eye. Change the distance between the objective lens of the microscope and the erbium-doped yttrium vanadate crystal, so that a blue-green visible light spot appears inside the cadmium sulfide crystal, and the blue-green visible light spot is the focus of the objective lens of the microscope. The distance A between them, or the distance B between the light source of the microscope and the stage, determines the position of the focal point of the objective lens of the microscope in space.
作为优选，所述的改变显微镜的物镜与转换发光晶体之间距离的方法为，保持显微镜的载物台以及载物台上的转换发光晶体固定不动，通过调节显微镜的镜筒升降，带动显微镜的物镜移动，以改变显微镜的物镜与转换发光晶体之间的距离。Preferably, the method for changing the distance between the objective lens of the microscope and the conversion luminescence crystal is to keep the stage of the microscope and the conversion luminescence crystal on the stage fixed, and adjust the lens barrel of the microscope to move up and down to drive the microscope The objective lens moves to change the distance between the microscope objective lens and the converted luminescent crystal.
作为优选，所述的改变显微镜的物镜与转换发光晶体之间距离的方法为，保持显微镜的物镜固定不动，通过调节显微镜的载物台升降，带动载物台上的转换发光晶体移动，以改变显微镜的物镜与转换发光晶体之间的距离。Preferably, the method for changing the distance between the objective lens of the microscope and the converted luminescent crystal is to keep the objective lens of the microscope fixed, and adjust the stage of the microscope to move the converted luminescent crystal on the stage to Change the distance between the objective lens of the microscope and the converted luminescent crystal.
本发明采用以上技术方案，在显微镜上应用转换发光晶体可将不可见激光转换为可见光的技术来确定显微镜的物镜焦点在空间上的位置，使得后期在使用显微镜观测物体时，只要将被测物体调节至显微镜物镜焦点所在焦平面位置，就可以快速、准确的调节出显微镜中被测物体的清晰图像，从而大大降低了显微镜焦点调节难度，节省了大量时间，降低了劳动强度，可以提高实验与分析的速度，降低人为操作对显微成像质量的影响，该方法操作简便，经济实用，适于推广。The present invention adopts the above technical scheme and applies the technique of converting invisible laser light into visible light on the microscope to determine the spatial position of the objective lens focus of the microscope, so that when the microscope is used to observe objects in the later stage, as long as the measured object By adjusting to the focal plane position where the focus of the microscope objective lens is located, a clear image of the object to be measured in the microscope can be quickly and accurately adjusted, which greatly reduces the difficulty of adjusting the focus of the microscope, saves a lot of time and labor intensity, and can improve the efficiency of experiments and The speed of analysis can reduce the influence of manual operation on the quality of microscopic imaging. The method is easy to operate, economical and practical, and is suitable for popularization.
附图说明Description of drawings
现结合附图对本发明作进一步阐述：Now in conjunction with accompanying drawing, the present invention is further elaborated:
图1为本发明显微镜物镜焦点位置确定原理示意图；Fig. 1 is the principle schematic diagram of determining the focus position of the microscope objective lens of the present invention;
图2为本发明实施例1中应用980nm激光照射掺铒钒酸钇晶体以确定显微镜物镜焦点的照片；图中掺铒钒酸钇晶体内绿色的可见光点即为显微镜的物镜焦点；Fig. 2 is the photograph that application 980nm laser irradiates erbium-doped yttrium vanadate crystal to determine the focal point of the microscope objective lens in the embodiment of the present invention 1; Among the figure, the visible light point of green in the erbium-doped yttrium vanadate crystal is the objective lens focus of microscope;
图3为本发明实施例1掺铒钒酸钇晶体光致发光光谱图。Fig. 3 is a photoluminescence spectrum diagram of erbium-doped yttrium vanadate crystal in Example 1 of the present invention.
具体实施方式Detailed ways
如图1-3之一所示，一种应用激光转换为可见光确定显微镜物镜焦点的方法，该方法是以不可见激光作为显微镜的光源1，将转换发光晶体2置于显微镜的载物台上，并对准显微镜的通光孔中央，改变显微镜的物镜3与转换发光晶体之间的距离，使不可见激光照射在转换发光晶体2上，在转换发光晶2体内聚焦出现有颜色的可见光点4，该可见光点4即为显微镜物镜焦点，该可见光点所在位置即为显微镜物镜焦点位置。As shown in one of Figures 1-3, a method of converting laser light into visible light to determine the focus of the microscope objective lens, the method uses invisible laser light as the light source 1 of the microscope, and places the converted luminescent crystal 2 on the stage of the microscope , and aim at the center of the light hole of the microscope, change the distance between the objective lens 3 of the microscope and the conversion luminescent crystal, so that the invisible laser light is irradiated on the conversion luminescence crystal 2, and a colored visible light point appears in the conversion luminescence crystal 2. 4. The visible light spot 4 is the focus of the microscope objective lens, and the position of the visible light spot is the focus position of the microscope objective lens.
作为优选，所述转换发光晶体为上转换发光晶体或者下转换发光晶体。Preferably, the conversion light-emitting crystal is an up-conversion light-emitting crystal or a down-conversion light-emitting crystal.
作为优选，所述的上转换发光晶体为掺铒钒酸钇晶体，所述的不可见激光的波长为980nm，所述的可见光点为绿色。Preferably, the up-conversion luminescent crystal is an erbium-doped yttrium vanadate crystal, the wavelength of the invisible laser is 980nm, and the visible light point is green.
进一步，所述的掺铒钒酸钇晶体采用提拉法制得，掺铒钒酸钇晶体的的掺铒浓度为0.2-1.2mol%。本发明采用掺铒浓度为0.2-1.2mol%的掺铒钒酸钇晶体使得绿光具有较长的荧光寿命，而且分辨率好，成本低。Further, the erbium-doped yttrium vanadate crystal is prepared by a pulling method, and the erbium-doped concentration of the erbium-doped yttrium vanadate crystal is 0.2-1.2 mol%. The invention adopts the erbium-doped yttrium vanadate crystal with the erbium-doped concentration of 0.2-1.2 mol%, so that the green light has a long fluorescence lifetime, good resolution and low cost.
作为优选，所述的下转换发光晶体为硫化镉晶体，所述的可见激光的波长为325nm，所述的可见光点为蓝绿色。Preferably, the down-conversion luminescent crystal is cadmium sulfide crystal, the wavelength of the visible laser is 325nm, and the visible light spot is blue-green.
作为优选，所述的改变显微镜的物镜与转换发光晶体之间距离的方法为，保持显微镜的载物台以及载物台上的转换发光晶体固定不动，通过调节显微镜的镜筒升降，带动显微镜的物镜移动，以改变显微镜的物镜与转换发光晶体之间的距离。Preferably, the method for changing the distance between the objective lens of the microscope and the conversion luminescence crystal is to keep the stage of the microscope and the conversion luminescence crystal on the stage fixed, and adjust the lens barrel of the microscope to move up and down to drive the microscope The objective lens moves to change the distance between the microscope objective lens and the converted luminescent crystal.
作为优选，所述的改变显微镜的物镜与转换发光晶体之间距离的方法为，保持显微镜的物镜固定不动，通过调节显微镜的载物台升降，带动载物台上的转换发光晶体移动，以改变显微镜的物镜与转换发光晶体之间的距离。Preferably, the method for changing the distance between the objective lens of the microscope and the converted luminescent crystal is to keep the objective lens of the microscope fixed, and adjust the stage of the microscope to move the converted luminescent crystal on the stage to Change the distance between the objective lens of the microscope and the converted luminescence crystal.
实施例1Example 1
1）、制备掺铒钒酸钇晶体：采用提拉法制得掺铒钒酸钇晶体，掺铒钒酸钇晶体的的掺铒浓度为0.2-1.2mol%。1) Preparation of erbium-doped yttrium vanadate crystals: the erbium-doped yttrium vanadate crystals were prepared by the pulling method, and the erbium-doped yttrium vanadate crystals had an erbium-doped concentration of 0.2-1.2 mol%.
2）、应用激光转换为可见光确定显微镜物镜焦点：方法是以波长为980nm的不可见激光作为显微镜的光源，将掺铒钒酸钇晶体作为上转换发光晶体置于显微镜的载物台上，并对准显微镜的通光孔中央，使不可见激光照射在掺铒钒酸钇晶体上，掺铒钒酸钇晶体内出现波长为553nm肉眼可见的绿色光子，保持显微镜的载物台以及载物台上的掺铒钒酸钇晶体固定不动，通过调节显微镜的镜筒升降，带动显微镜的物镜移动，以改变显微镜的物镜与掺铒钒酸钇晶体之间的距离，使掺铒钒酸钇晶体内聚焦出现绿色的可见光点，该绿色可见光点即为显微镜物镜焦点，此时，测量显微镜的物镜与载物台之间的距离A，从而确定了显微镜的物镜焦点在空间上的位置。2) Apply laser conversion to visible light to determine the focal point of the microscope objective lens: the method is to use an invisible laser with a wavelength of 980nm as the light source of the microscope, place the erbium-doped yttrium vanadate crystal as an up-conversion luminescent crystal on the stage of the microscope, and Align the center of the light hole of the microscope, so that the invisible laser is irradiated on the erbium-doped yttrium vanadate crystal, and green photons with a wavelength of 553nm visible to the naked eye appear in the erbium-doped yttrium vanadate crystal, keeping the microscope stage and the stage The erbium-doped yttrium vanadate crystal on the erbium-doped yttrium vanadate crystal is fixed, and the objective lens of the microscope is moved by adjusting the lens barrel of the microscope to change the distance between the microscope objective lens and the erbium-doped yttrium vanadate crystal, so that the erbium-doped yttrium vanadate crystal A green visible light spot appears in the inner focus, which is the focus of the microscope objective lens. At this time, measure the distance A between the objective lens of the microscope and the stage to determine the spatial position of the objective lens focus of the microscope.
3）、使用显微镜观测被测物体：方法是采用显微镜常规操作方法，保持显微镜的载物台固定不动，将被测物体置于显微镜的载物台上，通过调节显微镜的镜筒升降，带动显微镜的物镜移动，使得显微镜的物镜与载物台之间距离为A，从而使被测物体位于显微镜物镜焦点所在焦平面位置，进而可以快速、直观、准确的调节出显微镜中被测物体的清晰图像。3) Use a microscope to observe the measured object: the method is to use the conventional microscope operation method to keep the stage of the microscope fixed, place the object to be measured on the stage of the microscope, and adjust the lens barrel of the microscope to move up and down to drive The objective lens of the microscope moves so that the distance between the objective lens of the microscope and the stage is A, so that the measured object is located at the focal plane where the focus of the microscope objective lens is located, and then the clarity of the measured object in the microscope can be adjusted quickly, intuitively and accurately. image.
实施例2Example 2
1）、制备硫化镉晶体：采用提拉法制得硫化镉晶体。1) Preparation of cadmium sulfide crystals: The cadmium sulfide crystals were prepared by the pulling method.
2）、应用激光转换为可见光确定显微镜物镜焦点：方法是以波长为325nm的不可见激光作为显微镜的光源，将硫化镉晶体作为下转换发光晶体置于显微镜的载物台上，并对准显微镜的通光孔中央，使不可见激光照射在硫化镉晶体上，硫化镉晶体内出现肉眼可见的蓝绿色光子，保持显微镜的物镜固定不动，通过调节显微镜的载物台升降，带动载物台上的硫化镉晶体移动，以改变显微镜的物镜与硫化镉晶体之间的距离，使不可见激光照射在硫化镉晶体上，硫化镉晶体内聚焦出现蓝绿色的可见光点，该蓝绿色可见光点即为显微镜物镜焦点，此时，测量显微镜的光源与载物台之间的距离B，从而确定了显微镜的物镜焦点在空间上的位置。2) Apply laser conversion to visible light to determine the focus of the microscope objective lens: the method is to use an invisible laser with a wavelength of 325nm as the light source of the microscope, place a cadmium sulfide crystal as a down-conversion luminescent crystal on the stage of the microscope, and align it with the microscope In the center of the light hole of the cadmium sulfide crystal, the invisible laser is irradiated on the cadmium sulfide crystal, and the blue-green photons visible to the naked eye appear in the cadmium sulfide crystal, and the objective lens of the microscope is kept fixed. By adjusting the lifting of the stage of the microscope, the stage is driven The cadmium sulfide crystal on the cadmium sulfide crystal moves to change the distance between the objective lens of the microscope and the cadmium sulfide crystal, so that the invisible laser light is irradiated on the cadmium sulfide crystal, and a blue-green visible light point appears in the cadmium sulfide crystal. The blue-green visible light point is is the focal point of the objective lens of the microscope. At this time, the distance B between the light source of the microscope and the stage is measured, thereby determining the position of the focal point of the objective lens of the microscope in space.
3）、使用显微镜观测被测物体：方法是采用显微镜常规操作方法，保持显微镜的物镜固定不动，将被测物体置于显微镜的载物台上，通过调节显微镜的载物台升降，带动载物台上的被测物体移动，使得显微镜的光源与载物台之间的距离为B，从而使被测物体位于显微镜物镜焦点所在焦平面位置，进而可以快速、直观、准确的调节出显微镜中被测物体的清晰图像。3) Use a microscope to observe the measured object: the method is to use the conventional microscope operation method to keep the objective lens of the microscope fixed, place the measured object on the stage of the microscope, and adjust the lifting of the stage of the microscope to drive the load The object to be measured on the stage moves so that the distance between the light source of the microscope and the stage is B, so that the object to be measured is located at the focal plane where the focal point of the microscope objective lens is located, and then it can be adjusted quickly, intuitively and accurately. A clear image of the object being measured.
以上描述不应对本发明的保护范围有任何限定。The above description should not limit the protection scope of the present invention in any way.
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