Fluorescent Leukocytes Enter Plaque on the Microscope Stage
Atherosclerosis is a disease driven by lipid accumulation and leukocyte influx in the vessel wall.1 Because visualizing leukocyte behavior in vivo is the holy grail for those interested in understanding leukocyte migration, major progress has been made in developing intravital microscopy approaches to study leukocyte movement. Understanding how certain leukocytes, such as T cells, participate in various diseases including cancer2,3 has been one of intravital microscopy’s success stories. The technology has also proven beneficial for understanding basic processes of leukocyte biology, such as the patrolling behavior of monocytes,4 monocyte movement within the spleen,5 the existence of neutrophil slings,6 and the differentiation of hematopoietic stem cells to leukocytes in the bone marrow.7 These 3-dimensional methods that combine high temporal resolution, (sub)cellular spatial resolution, and multicolor encoding for multiple cells or proteins can inform on in vivo biology in unprecedented ways, including real-time interaction of cells with each other and the tissue environment.
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One of the major challenges in performing intravital microscopy in live animals is the need to control respiratory, peristaltic, and cardiovascular motion. The beating heart (and to a somewhat lesser degree, the arterial vasculature) in a live animal moves rapidly with a high amplitude of motion. Without sophisticated stabilization methods, cardiovascular motion makes it impossible to focus on a cell residing in tissue, let alone isolation and resolution …