THE STREAM LINE™

The Imaging in Flow newsletter from Amnis® Corporation

The Stream Line was designed to keep you up to date with the applications and technical developments related to the Amnis ImageStream® System, the revolutionary new technology for imaging in flow. You will also find information on how to contact Amnis and where you will be able to see ImageStream technology firsthand. For further information, please visit the Amnis website at www.amnis.com

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Application Note: Analysis of the Nuclear Translocation of NF-kB using ImageStream Multispectral Imaging Cytometry.

Aberrant regulation of NF-kB transcription factors has been implicated in chronic and acute inflammatory diseases, autoimmune diseases and some types of cancer. Because of this widespread disease involvement, activation of the NF-kB signaling pathway, including translocation of NF-kB from the cytoplasm to the nucleus, has been an important subject of investigation in basic research and drug discovery. Conventional flow cytometry has offered little to the analysis of this problem, however, as the technology cannot provide important cellular imagery. The Amnis ImageStream System, in contrast, not only delivers the flow cytometric data necessary to analyze the wide range of cellular responses, but also provides up to six high resolution images of every cell. The combination of flow cytometry and microscopy in a single instrument enables an entirely new approach to the analysis of problems such as NF-kB translocation.

In this experiment, adherent A549 cells and non-adherent THP-1 cells were each stimulated to activate the NF-kB signaling pathway and p65 translocation was followed with fluorescent labeled anti-NF-kB. The stain 7-AAD was used to identify the nucleus, and in addition brightfield and darkfield images were taken of every cell. Each cell population was analyzed by traditional fluorescence microscopy as well as with the ImageStream 100. On the ImageStream system, the extent of nuclear translocation was quantified for every cell in the population by determining the degree of similarity in the localization of staining between the cell nucleus and the NF-kB. Results showed clearly measurable translocation in both adherent and non-adherent cells after stimulation. Moreover, the population data provided valuable information on the fraction and type of cells in which significant translocation was observed.

Click here to download a PDF poster presentation of this experiment.

Visit Amnis at the International Society for Analytical Cytology XXII Montpellier, France May 23-27

If you will be attending ISAC XXII this month, stop by the Amnis booth (44-45) to get a close look at the ImageStream System and powerful IDEAS(TM) analytical software. And be sure to catch one or more of the presentations and posters covering ImageStream technology:

Presentations

IMAGING IN FLOW: EARLY APPLICATION DATA WITH THE IMAGESTREAM 100 BETA PROTOTYPE

Gary Elliott, PhD Resesarch Scientist Flow Cytometry, Amgen Inc

Parallel Session I Sunday, May 23 3:00-4:45PM

CHARACTERISTICS AND APPLICATIONS OF IMAGESTREAM MULTISPECTRAL IMAGING FLOW CYTOMETRY

David Basiji, PhD Chief Technology Officer, Amnis Corporation

Parallel Session 6 Wednesday, May 26 8:00 - 9:45AM

Posters

Abstract #97084 MULTISPECTRAL HIGH CONTENT CELLULAR ANALYSIS USING A FLOW BASED IMAGING CYTOMETER

Abstract #96980 QUANTITATION OF NUCLEAR TRANSLOCATION EVENTS USING IMAGESTREAM MULTISPECTRAL IMAGING CYTOMETRY

Abstract #96527 DISTINGUISHING MODES OF CELL DEATH USING IMAGESTREAM MULTISPECTRAL IMAGING CYTOMETRY

Abstract #97095 FLUORESCENT IN SITU HYBRIDIZATION IN SUSPENSION ANALYSIS USING IMAGESTREAM MULTISPECTRAL IMAGING FLOW CYTOMETRY