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Immunofluorescent staining using antibodies to detect specific proteins allows for visualization of proteins of interest in a biological sample. In recent years, there have been important advances in the microscopy equipment used for imaging, and we can now perform so-called high-resolution microscopy. Through high-resolution microscopy we can not only study biological processes but also visualize them.

Imaging through the IVIS Spectrum CT system does not provide the resolution at cellular level like the high-resolution or super-resolution microscopy. Rather, it detects bacterial infections in specific anatomical compartments/organs of the animals. The IVIS Spectrum imaging system is a unique imaging technology that allows for real-time monitoring of disease progression in living animals through the use of either bioluminescent or fluorescent probes.

In vivo imaging, meaning imaging tissues in living animals, is still a developing technique. However, microscopy imaging ex vivo remains a very important tool that allows for visualization of biological and pathological processes occurring in vivo. As described in Chap. 5, imaging of animal and human tissue postmortem can be performed at high resolution. Recently, imaging of human tissues infected with pneumococci using an even higher resolution, the so-called super-resolution with STED, has been reported.

Animal models are fundamental tools to study the biology of physiological processes and disease pathogenesis. To study invasive pneumococcal disease (IPD), many models using mice in particular have been established and developed during recent years. Thanks to the advances of the research in the pneumococcal field, nowadays, there is the possibility to use defined mouse models to study each disease caused by the pneumococcus. In this chapter mouse models for pneumonia, bacteremia, and meningitis are described. Since pneumococci are commensal pathogens found to a high extent in healthy individuals. Hence, we also describe a mouse model for nasopharyngeal colonization.