散斑照明宽场荧光层析显微成像技术研究

介绍了一种新的宽场荧光层析显微方法.在传统宽场显微镜中引入散斑图案照明样品，控制散斑图案的动态变化，利用CCD相机记录对应的一系列荧光图像.由于焦平面内强度变化远比焦平面外强度变化剧烈，通过合适的算法能够获得焦平面的层析分辨的荧光显微图像.标定了系统参数，并研究了不同的图像重建算法对系统性能的影响，获得了不同生物组织样品的层析图像.实验表明，该显微方法能用于组织光学切片成像，在临床医学中具有实际应用价值. 关键词： 荧光 散斑照明 荧光显微 层析

Study on wide-field fluorescence sectioning microscopy based on dynamic speckle illumination

In this paper, we present a novel wide-field fluorescence sectioning microscopy in which speckle patterns are produced on the sample for illumination. The speckle pattern is dynamically changed and a sequence of fluorescence images of the sample is recorded with a CCD camera. Due to a large variation of fluorescence intensity of the in-focus light compared to that of the out-of-focus light, special processing algorithms can be used to reconstruct the sectioned fluorescence image. We calibrated the system and studied the effect of the reconstruction algorithms on the system performance. Fluorescence sectioned images of a few biological samples were obtained. Our experiments showed that this wide-field fluorescence sectioning microscopy can be used to optically section tissue and has potential applications in the clinics.