Abstract: A fluorescence microscope system including an optical detection system configured to capture a raw image of a sample, the raw image including a plurality of pixels, each pixel having a brightness value and a processor, configured to determine one or more invalid pixels in the raw image, assign a predetermined value to each invalid pixel, determine a range of brightness values including the brightness values of a majority of the plurality of pixels excluding the one or more invalid pixels, and generate a processed image of the sample based on the determined range of brightness values. (FIG.

Abstract: An image processing apparatus for determining a focused output image in a passive autofocus system is configured to retrieve a set of input images and compute a baseline estimate for at least one input image. The baseline estimate represents image structures in the input image. The image structures have a length scale larger than a predetermined image feature length scale. The image processing apparatus is further configured to compute a set of output images, wherein each output image of the set of output images is computed based on one of a different input image of the set of input images and the at least one baseline estimate for the different input image and the at least one baseline estimate for a respective different input image. The image processing apparatus is further configured to determine one output image of the set of output images as the focused output image.

Abstract: A microscope includes illumination optics for fluorescence excitation of point light sources of a sample, detection optics and a camera having a sensor. A density of the point light sources is kept low so as to minimize a crossover of point light sources that are behind or close to one another in each image captured by the camera. A means for subdividing a detection aperture into individual sub-apertures is provided in a beam path of the detection optics such that images generated by the individual sub-apertures on the sensor of the camera depict an object volume from different spatial directions.

Abstract: A microscope includes illumination optics for fluorescence excitation of point light sources of a sample, detection optics and a camera having a sensor. A density of the point light sources is kept low so as to minimize a crossover of point light sources that are behind or close to one another in each image captured by the camera. A means for subdividing a detection aperture into individual sub-apertures is provided in a beam path of the detection optics such that images generated by the individual sub-apertures on the sensor of the camera depict an object volume from different spatial directions.