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共聚焦显微镜的主分光装置
当前荧光显微镜采用入射照明，这需要将照明和发射光分开。传统的分光设备是一个颜色依赖的分光镜，它具有固定的光谱参数，并通常在指定的波长带内透射90%至98%的发射光。透射率依赖于波长，但也受到技术、设计和实验需求的影响。声光光束分割器（AOBS）则是一种可自由调节的反射缺口设备，平均95%的发射光在这些狭窄的缺口间透射。
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![[Translate to chinese:] Pinhole diameter and diffraction pattern.](/fileadmin/_processed_/9/f/csm_Pinhole_diameter_and_diffraction_pattern_a8f1bc8eca.jpg "[Translate to chinese:] Pinhole diameter and diffraction pattern.")
共聚焦显微镜针孔效应
在操作共聚焦显微镜，或在讨论这种装置的特性和参数时，我们不可避免地提到针孔及其直径。这篇简短的文章是针对那些没有足够时间钻研共聚焦显微镜的理论和细节但又想了解针孔效应的用户们来解释针孔的意义。
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![[Translate to chinese:] Left: Tissue cells marked with an immunolabel (FITC) illuminated with wide-band UV excitation. Note the tissue structure with blue autofluorescence. Right: Same tissue and same immunostaining with FITC label illuminated with epi-illumination using narrow-band blue (490 nm) light. Note the increased image contrast (Ploem, 1967)](/fileadmin/_processed_/c/2/csm_Ploem_Figure_5_Autofluorescence_a_b_3ad909dc27.png "[Translate to chinese:] Left: Tissue cells marked with an immunolabel (FITC) illuminated with wide-band UV excitation. Note the tissue structure with blue autofluorescence. Right: Same tissue and same immunostaining with FITC label illuminated with epi-il")
Milestones in Incident Light Fluorescence Microscopy
Since the middle of the last century, fluorescence microscopy developed into a bio scientific tool with one of the biggest impacts on our understanding of life. Watching cells and proteins with the…
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Milestones in Incident Light Fluorescence Microscopy
Since the middle of the last century, fluorescence microscopy developed into a bio scientific tool with one of the biggest impacts on our understanding of life. Watching cells and proteins with the…
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BABB Clearing and Imaging for High Resolution Confocal Microscopy
Multipohoton microscopy experiment using Leica TCS SP8 MP and Leica 20x/0.95 NA BABB immersion objective.
Understanding kidney microanatomy is key to detecting and identifying early events in kidney…
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From Light to Mind: Sensors and Measuring Techniques in Confocal Microscopy
This article outlines the most important sensors used in confocal microscopy. By confocal microscopy, we mean "True Confocal Scanning", i.e. the technique that illuminates and measures one single…
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![[Translate to chinese:] Acousto-optics, sketch](/fileadmin/_processed_/f/5/csm_Acousto-optics_sketch_01_67e5a72752.jpg "[Translate to chinese:] Acousto-optics, sketch")
声光调制在全光谱型激光共聚焦显微镜系统的应用
荧光最显著的特征是照射光（激发光）和检测光（发射光）颜色之间的偏移，称为斯托克斯位移。因此，在荧光成像中，不仅要将激发光和发射光的相应波长过滤出来，还需要将激发光从发射光中分离。过去，通常用平面光学元件，包括灰色或彩色滤光片和反射镜进行滤光和分光。虽然有多种平面光学元件可供使用，但固定的规格和低切换效率使其在使用上具有局限性，并且采用不同角度或梯度的涂层作为激发光和发射光的调谐方法也被证实并不可行…
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光谱检测-如何设定特定探针发射光的光谱检测范围
为了拆分多色成像的发射光谱，首先由分束器或色散元件将不同颜色的光引入到不同的方向[1]，带通滤片则能够最大限度地减少串色，并抑制所有残留的激发光，最终到达传感器。在过去，常使用的滤片是普通的玻璃带通滤片。如今，一项革命性的设计诞生了，那就是在多波段组件（SP探测器）中使用光度计滑块。该设计可以极为有效地探测发射光，同时提供完全可调谐性，与此同时带来的好处是使光谱扫描成为了可能。使用白激光作为光源时…
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FRAP实验步骤式指南
漂白后荧光恢复（FRAP）已被认定为观察大分子平移扩散过程方面使用最为广泛的一种方法。由此产生的信息可用于确定动力学性质，如扩散系数、流动分数和荧光标记分子的传输速率。FRAP实验利用了短激光脉冲的荧光团辐照。先进的激光扫描显微镜如TCS…