M182:Caracino:Neutrophil Adhesion: A HCS Compatible Assay Using the Acumen eX3

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Diana Caracino and Paul Wylie

A variety of pathological conditions exist in which cellular adhesion events are key to the evolving disease state. These include myocardial infarction, Adult Respiratory Distress Syndrome (ARDS), and general trauma. Cell adhesion also plays a crucial role in the immune system and is an important factor in inflammatory diseases such as rheumatoid arthritis, asthma and psoriasis. The concepts involved in the cell adhesion process are a rapidly developing area of cell biology and over the last five to ten years it has become possible to work out, at a molecular level, how cells attach to each other and to extracellular matrix molecules. This will be important for future drug development. The importance of cell adhesion molecules in the homing of lymphocytes to lymphoid organs, in neutrophil localization in inflammation, and in the interaction of both lymphocytes and neutrophils with vascular endothelium suggests that defects in these molecules might have severe consequences.
The Acumen eX3 is the fastest imaging system available, collecting and simultaneously analysing over 40 images/second, covering the entire well, without the trade off of having to use lower resolution. It offers up to three lasers from a range between 405nm to 633nm up to 4 channels of data collection per laser. The Acumen eX3 can be used to study the process of cellular adhesion, whereby adherent cells types specifically, endothelial cells can be grown to confluence in microtitre plate wells and other cells types e.g. neutrophils added. The neutrophils can be differentially stained with calcein AM and the adhesion profile monitored and quantified. Cell adhesion can be determined simply by correlating retained fluorescence with cell number. It is envisaged that this type of assay will be of use in the pharmaceutical screening of novel drugs directed against specific cellular adhesion molecules and in academia for the monitoring of the adhesion process in different disease states, such as myocardial infarction, Adult Respiratory Distress Syndrome (ARDS) and stroke in vitro. . A simple, rapid, quantitative adhesion assay, routinely returning a signal to noise value of 6 to 1 has now been developed for the Acumen eX3 laser.

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