PROPERTIES OF A 4PI CONFOCAL FLUORESCENCE MICROSCOPE
PROPERTIES OF A 4PI CONFOCAL FLUORESCENCE MICROSCOPE
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DOI：
10.1364/josaa.9.002159
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发表时间：
1992-12-01
影响因子：
1.9
通讯作者：
STELZER, EHK
中科院分区：
文献类型：
--
作者：
HELL, S;STELZER, EHK
在4Pi共焦荧光显微镜中，使用两个相对的显微镜物镜从两侧照射荧光物体并收集两侧的荧光发射。无论是在共同的焦点或共同的检测器针孔中的检测波前的照明波前的建设性干涉导致的轴向分辨率比在共焦荧光显微镜高约4倍。建立了一个精确的4Pi共聚焦荧光显微镜，使用同时照明。当使用具有1.4的数值孔径、633 nm的激发和大约725 nm的检测的透镜时，深度辨别的半最大值处的全宽度通过实验确定为大约110 nm。
In a 4Pi confocal fluorescence microscope two opposing microscope objective lenses were used to illuminate a fluorescent object from both sides and to collect the fluorescence emissions on both sides. Constructive interference of either the illumination wave fronts in the common focus or the detection wave fronts in the common detector pinhole resulted in an axial resolution approximately four times higher than that in a confocal fluorescence microscope. A precise 4Pi confocal fluorescence microscope that uses simultaneous illumination was built. The full width at half-maximum of the depth discrimination was determined experimentally to be approximately 110 nm when lenses with a numerical aperture of 1.4, an excitation of 633 nm, and detection of approximately 725 nm were used.