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This textbook (or extended technology sheet) is based on more than ten years of experience of the Institute of Complex Systems (ICS) FFPW USB in light microscopy and digital camera-based measurements. The textbook presents the theoretical and related applied results of the Laboratory of Experimental Complex Systems. The authors published original theoretical findings in several international scientific works from 2013–2021. The authors carried out all presented measurements on custom-made microscopes assembled in cooperation with Czech and Austrian engineering companies: Petr Tax-Optax (Praha) and Synchronics Engineering (Heidenreichstein) helped us build the microscope hardware. Petr Mach´aˇcek-ImageCode (Brloh) is responsible for translating the microscope control software and other relevant data processing software. The textbook contains two main chapters – theoretical and experimental. The goal of this textbook is for students to understand that if they see something under a microscope, it may not be real. The first part of theoretical Chapter 1 is based on Tom´aˇs N´ahl´ık’s dissertation in the field of Biophysics at the Faculty of Science USB (N´ahl´ık, 2016). In this part, students will learn about imaging principles in light microscopy. Some possible optical system aberrations from the point of view of image digitisation are also worth noting. The presented figures arose from the computational application of Extended Nijboer-Zernike models (Braat et al., 2015) and our experimental measurements. In the second part of Chapter 1, we introduce original information entropy methods that we further implemented as software tools for processing and analysing a digital image obtained by light microscopy. Chapter 1 ends by test questions verifying student’s final knowledge.