Microscopy Fundamentals

This chapter provides an introduction to the microscope, and focuses on critical aspects of microscopy that impact high content screening (HCS), such as the practical limits of resolution, spectral bandwidth and the ability to multiplex several wavelengths. Every commercialized design of HCS imager has many specific elements to its use and design that strike a balance between flexibility and ease of use. Historically, a discussion of microscopy fundamentals has combined the factors that influence the quality of the science with the care and maintenance of the microscope itself. The chapter discusses some types of HCS microscopy (fluorescence microscopy, confocal microscopy, brightfield microscopy, and live‐cell Imaging) that entail specific instrument configurations. It describes each of the components of the HCS microscope, as applied to HCS. HCS platforms usually capture images through charge‐coupled device (CCD) or complementary metal oxide semiconductor (CMOS) cameras, while pointscanning confocal HCS platforms use a photomultiplier tube (PMT).