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.