![]() In this review, we explore the growing field of microfluidics and present an up-to-date overview of the different approaches and techniques used for single-cell applications in immunology. Single-cell technologies present considerable advantages that can overcome the limitations of bulk measurements and can help achieve a better understanding of immune mechanisms, which in turn is expected to lead to efficient, personalized immune treatments for complex illness like autoimmune diseases and cancer. These unique features are often missed by conventional, bulk measurements that usually provide population level averages. Cells from the adaptive immune system (T and B cells mostly) are genetically unique due to VDJ recombination and clonal selection. For instance, macrophages can present several different phenotypes, ranging from anti- to pro-inflammatory. In addition to the diversity of cell types constituting the immune system, each cell type can present distinct features or can be genetically unique. The immune system is a complex system consisting of a variety of cell types that work in synergy to protect against invading pathogens and control infected and mutated cells. This success can be explained by various advantages microfluidic based approaches have over conventional technologies such as precise spatio- and temporal control of extremely small volumes, reducing costs and required sample volumes, while providing sensitivity and throughput. Microfluidics significantly evolved over several decades and it is now used in almost all biological fields including biochemistry, cell signaling, drug testing, genomics, and proteomics. Microfluidics, the science of manipulating fluids on the microscale, has had considerable impact on biology, both in research and industry. In this review, we give an overview of state-of-the-art microfluidic techniques, their application to single-cell immunology, their advantages and drawbacks, and provide an outlook for the future of single-cell technologies in research and medicine. Microfluidics has been used in a large panel of biological applications, ranging from single-cell genomics, cell signaling and dynamics to cell–cell interaction and cell migration studies. In addition, microfluidic devices can achieve high-throughput measurements, enabling in-depth studies of complex systems. These features are highly desirable for single-cell analysis and have made microfluidic devices useful tools for studying complex immune systems. By controlling device geometries, surface chemistry, and flow behavior, microfluidics can create a precisely defined microenvironment for single-cell studies with spatio-temporal control. Microfluidic systems enable the precise control of small volumes in the femto- to nanoliter range. Advances in the fields of microsystems and microengineering gave rise to the field of microfluidics and its application to biology. There is a need in immunology to measure single cells and to study the dynamic interplay of immune cells with their environment. Conventional bulk methods are often prone to miss important details by returning population-averaged results. ![]() Immune cells are highly diverse, and an immune response is the result of a large number of molecular and cellular interactions both in time and space. Many of the mechanisms and their regulation remain poorly understood. Imbalances in this network often result in excessive or absent immune responses leading to allergies, autoimmune diseases, and cancer. The immune system is a complex network of specialized cells that work in concert to protect against invading pathogens and tissue damage. ![]()
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