The ultimate aim of fluorescence microscopy is to achieve high-resolution imaging of increasingly larger biological samples. Extended depth of field presents a potential solution to accelerate imaging of large samples when compression of information along the optical axis is not detrimental to the interpretation of images. We have implemented an extended depth of field (EDF) approach in a random illumination microscope (RIM). RIM uses multiple speckled illuminations and variance data processing to double the resolution. It is particularly adapted to the imaging of thick samples as it does not require the knowledge of illumination patterns. We demonstrate highly-resolved projective images of biological tissues and cells. Compared to a sequential scan of the imaged volume with conventional 2D-RIM, EDF-RIM allows an order of magnitude improvement in speed and light dose reduction, with comparable resolution. As the axial information is lost in an EDF modality, we propose a method to retrieve the sample topography for samples that are organized in cell sheets.

荧光显微镜的最终目标是实现越来越大的生物样品的高分辨率成像。当沿光轴压缩信息不会影响图像的解释时，扩展景深提供了一种潜在的解决方案，可以加速大型样品的成像。我们在随机照明显微镜 （RIM） 中实施了扩展景深 （EDF） 方法。RIM 使用多个斑点照明和方差数据处理将分辨率提高一倍。它特别适用于厚样品的成像，因为它不需要照明模式的知识。我们展示了生物组织和细胞的高分辨率投影图像。与使用传统 2D-RIM 对成像体积进行顺序扫描相比，EDF-RIM 可以在分辨率下将速度和光剂量减少提高一个数量级。由于轴向信息在 EDF 模态中丢失，我们提出了一种方法来检索以细胞片形式组织的样品的样品地形。