Abstract
Leukocytes are among the most mobile and versatile cells that have many essential functions in homeostasis and survival. Especially cells from the innate immune system, i.e., neutrophils and macrophages, play an important role as rapid first responders against invading microorganisms. With the advent of novel imaging techniques, new ways of visualizing innate immune cells have become available in recent years, thereby enabling more and more detailed discoveries about their nature, function and interaction partners. Besides intravital spinning-disc and 2-photon microscopy, clearing and 3D-imaging techniques provide new insights into the mechanism of innate immune cell behavior in their natural environment. This mini review focuses on the contributions of novel-imaging techniques to provide insight into the functions of neutrophils and macrophages under homeostasis and in infections. Imaging setups for different organs like the liver, kidney, heart, lung, and the peritoneal cavity are discussed as well as the current limitations of these imaging techniques.
Highlights
The immune system is composed of many different cell types distributed throughout the body
Regardless of the microscopy techniques, another important point to always keep in mind is that the dark matter matters and can make up a big portion of the organs and cells we look at, as we can only see what is labeled or what is auto-fluorescent
Since the formation and dissolution of such interactions can be very fast, visualization requires high-imaging speed that can be provided by stateof-the-art spinning-disk intravital microscopy (SD-IVM) or microscopes using very fast scanners
Summary
The immune system is composed of many different cell types distributed throughout the body. Intravital microscopy had a huge impact on our understanding of the immune system as it is the only technique that allows to simultaneously study structure and function at a cellular to subcellular level with high spatial and temporal resolution in vivo. Since imaging techniques have developed a lot and nowadays it is possible to visualize the immune processes taking place in almost every organ—brain, eye, lung, heart, lymph node, joints, spleen, liver, gut, kidney, bladder, peritoneal cavity, mammary ducts and more using experimental mouse, or rat models. The development of better and faster microscopy techniques had a tremendous impact. We moved from simple light microscopy to more complex techniques. Scanning the specimen point by point slows the process down significantly. A spinning-disc setup consists of multiple pinholes on a rotating disc which speeds up the scanning process significantly thereby allowing to capture fast events like bacteria or platelets moving in the blood
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