细胞功能测定

Cell migration is required for a wide variety of processes from bacteria seeking for food to correct patterning of neuronal networks. The ability to sense external cues is critical for cells to get directions and reach their goals. So far, studies on chemotaxis have mainly focused their attenti ...

Migration of cells across endothelial barriers, termed transendothelial migration (TEM), is an important cellular process that underpins the pathology of many disease states including chronic inflammation and cancer metastasis. While this process can be modeled in vitro using c ...

Cell migration on two-dimensional (2D) substrates follows entirely different rules than cell migration in three-dimensional (3D) environments. This is especially relevant for leukocytes that are able to migrate in the absence of adhesion receptors within the confined geometry of ...

The endothelial cells lining blood vessels are continuously exposed to fluid shear stress generated by pulsatile flow of blood. In order to minimise forces acting on their surface, endothelial cells adapt to shear stress by alignment and migration within the direction of flow. Failure to ad ...

This chapter describes a method to study cells migrating in micro-channels, a confining environment of well-defined geometry. This assay is a complement to more complex 3D migration systems and provides several advantages even if it does not recapitulate the full complexity of 3D migrati ...

Cell migration is a process that is controlled by the formation and correct localization of protein complexes and by post-translational modification of individual proteins. Forster or fluorescent resonance energy transfer (FRET) detected using fluorescence lifetime imagi ...

This chapter describes the use of microscope-based fluorescence recovery after photobleaching (FRAP). To quantify the dynamics of proteins within a subcellular compartment, we first outline the general aspects of FRAP experiments and then provide a detailed protocol of how to measu ...

Cell migration, essential in cancer progression, is a complex process comprising a number of spatiotemporally regulated and well-coordinated mechanisms. In order to study (random) cell migration in the context of responses to various external cues (such as growth factors) or intrins ...

Total internal reflection fluorescence microscopy (TIRF-M) has become an increasingly popular tool to study events in close proximity to the cell cortex, such as cell adhesion (Axelrod, J Cell Biol 89:141–145, 1981; Gingell et al., J Cell Biol 100:1334–1338, 1985; Patel et al., J Cell Sci 121:1159–11 ...

The chick embryo is easily accessible and has therefore been widely used in developmental biology studies. In particular, the early embryo can be removed from the egg and cultured, which allows real-time observations and imaging. Here, we describe ex vivo electroporation followed by long- ...

Angiogenesis is the formation of new blood vessels from pre-existing vessels. This process is essential during embryonic development, wound healing, and regeneration as well as during several pathological conditions such as cancer. In this chapter, we describe an assay to measure angi ...

In the last decade, intravital microscopy on breast tumours in mice at single-cell resolution has resulted in important new insight into mechanisms of metastatic behaviour such as migration, invasion, and intravasation of tumour cells; angiogenesis; and the response of immune cells. T ...

Many steps of the metastatic cascade can be reproduced in simple in vitro assays such as tumour cell interactions with matrix proteins, proteolysis, chemotaxis, haptotaxis, and invasion into matrices or explanted tissues. Nevertheless, there are no fully adequate substitutes for the c ...

Studies using the social amoeba Dictyostelium discoideum have greatly contributed to the current understanding of the signaling network that underlies chemotaxis. Since directed migration is essential for normal D. discoideum multicellular development, mutants with che ...

Cell migration is a fundamental process that controls morphogenesis and inflammation. Its deregulation causes or is part of many diseases, including autoimmune syndromes, chronic inflammation, mental retardation, and cancer. Cell migration is an integral part of the cell biology, ...

Border cell migration in the Drosophila ovary has emerged as a genetically tractable model for studying collective cell movement. Over many years border cell migration was exclusively studied in fixed samples due to the inability to culture stage 9 egg chambers in vitro. Although culturing ...

A key feature of inflammatory cells is the ability to migrate to a site of injury or infection quickly and efficiently. Infectious agents can then be taken up by these inflammatory cells, preventing established infection. Inflammatory cell migration is driven by a complex interaction betw ...

This protocol describes an in vivo assay for random and directed hemocyte migration in Drosophila. Drosophila is becoming an increasingly powerful model system for in vivo cell migration analysis, combining unique genetic tools with translucency of the embryo and pupa, which allows di ...

The nematode Caenorhabditis elegans is an excellent model system in which to study long-distance cell migration in vivo. This chapter describes methods used to study a subset of migratory cells in the hermaphrodite nematode, the distal tip cells. These methods take advantage of the organis ...

Organotypic cultures are in vitro models that can be used to study the interactions between tumour and stromal cells. Collective tumour cell invasion in organotypic assays resembles that seen in human tissues in vivo, suggesting physiological relevance. A qualitative, pathological ...