Microscope Image Processing, Second Edition, introduces the basic fundamentals of image formation in microscopy including the importance of image digitization and display, which are key to quality visualization. Image processing and analysis are discussed in detail to provide readers with the tools necessary to improve the visual quality of images, and to extract quantitative information. Basic techniques such as image enhancement, filtering, segmentation, object measurement, and pattern recognition cover concepts integral to image processing. In addition, chapters on specific modern microscopy techniques such as fluorescence imaging, multispectral imaging, three-dimensional imaging and time-lapse imaging, introduce these key areas with emphasis on the differences among the various techniques. The new edition discusses recent developments in microscopy such as light sheet microscopy, digital microscopy, whole slide imaging, and the use of deep learning techniques for image segmentation and analysis with big data image informatics and management. Microscope Image Processing, Second Edition, is suitable for engineers, scientists, clinicians, post-graduate fellows and graduate students working in bioengineering, biomedical engineering, biology, medicine, chemistry, pharmacology and related fields, who use microscopes in their work and would like to understand the methodologies and capabilities of the latest digital image processing techniques or desire to develop their own image processing algorithms and software for specific applications.