T116: Advantages of acoustic focusing in flow cytometry and state of the art microscopy on your bench

Abstract

The work describes advantages of our latest instrument developments in flow cytometry and microscopy. Characteristics of newly launched flow cytometer, which uses acoustic focusing to align the cells in the focus of the laser for analysis, allows to analyze samples at a much higher throughput rate without losing precision and sensitivity. This opens up new possibilities regarding sample preparation and analysis. These advantages we want to demonstrate on 2 examples, whole blood analysis and rare event analysis. Analysis of biological samples in the most physiologic state with minimal sample preparation and manipulation is a key objective to any workflow. However as whole blood samples generally require significant manipulation, such as wash/centrifugation steps and/or red blood cell lysis, we have developed multiple no-lyse, no-wash assays to minimize these manipulation steps to obtain more physiological results. Here we show how we utilize the rapid sample collection capabilities of the our instrument to characterize phagocyte function in human whole blood with a phagocytosis/phagosome acidification assay and also a dihydrorhodamine 123 superoxide production assay, in a no-lyse, no wash format. Another area where our technology can shift boundaries is the field of rare event analysis. Due to the low frequency of the target cell population a large number of total events (in the range of several millions) need to be acquired. Simply due to the enormous amount of data that needs to be acquired, time becomes a very limiting factor for rare event analysis. As Acoustic Focusing allows analyzing samples at an approximately 10× higher throughput rate without scarifying data quality and maintaining a low coincidence rate, it pushes the limits what can be done in this field. As an example we will show the characterization of human iNKT cells. In addition to flow cytometry we want to show how modern microscopes help to perform a variety of routine and specialty applications.