The field of histopathology has encountered a key transition point with the progressive move towards use of digital slides and automated image analysis approaches. This chapter discusses the various methods and techniques involved in the automation of image analysis in histopathology. Important concepts and techniques are explained in the 5 main areas of workflow within image analysis in histopathology: data acquisition, the digital image, image pre-processing, segmentation, and machine learning. Furthermore, examples of the application of these concepts and techniques in histopathological research are then given.