Luminex ImageStreamX Mark II

Image StreamX

By combining the speed, sensitivity, and phenotyping abilities of flow cytometry with the detailed imagery and functional insight of microscopy, the ImageStreamX overcomes the limitations of both techniques and opens the door to an extensive range of applications: follow translocation from the cytoplasm to the nucleus; quantify capping and co-capping reactions; separate cells based on unique morphologic criteria; study cell-cell interactions; differentiate cell states such as late apoptosis and necrosis, study cell signaling, co-localization, shape change, internalization… – the range of potential applications is extremely broad.

You can define and quantify cell subpopulations not only by signal intensity, but by the location of fluorescent probes on, in, or between cells, even in rare sub-populations and highly heterogeneous samples. You can use cell and nuclear morphologies to characterize cell populations.

In the ImageStreamX, cells are illuminated with both a brightfield source and 405nm, 488nm, 560nm and 658nm lasers. It produces up to 12 high-resolution images (brightfield, darkfield and up to 10 fluorescence parameters) of each cell directly in flow, at rates up to 5,000 cells per second.

The IDEAS software package’s numerous pre-defined fluorescence and morphologic parameters let you quantitate virtually anything you can see so you can take full advantage of complex visual data to perform statistically-robust, multi-parameter quantitative analyses.

Applications that require the enumeration of spots within the cell or nucleus, like fluorescence in situ hybridization (FISH), require an increased depth of field. The ImageStreamX’s Extended Depth of Field (EDF) upgrade is a combination of specialized optics and unique image processing algorithms that project all structures and probes within the cell into one crisply-focused image. EDF makes it possible to find and count spots anywhere within the cell without any sacrifice in sensitivity or speed.

New features:

  • The Mark II upgrade provides real-time compensation, plotting, gating, and the use of gated events as data storage criteria. It accepts sample volumes from 20 to 200μL with 95% sample utilization thanks to its new fluidic system. It also sports a redesigned, intuitive user interface.
  • The new MultiMag option provides 20X and 60X objective lenses in addition to the standard 40X lens for greater experimental flexibility and improved resolution. The 60X objective increases magnification for small objects such as yeast and bacteria and offers greater detail with mammalian and plant cells. The 20X objective provides an increased field of view for the imaging of large cells, such as epithelial cells.
  • The 96-well plate AutoSampler option for the ImageStreamX imaging flow cytometer enhances productivity with unattended sample loading from multiwell plates. The fully integrated AutoSampler option allows you to easily perform dose response and time course studies even in primary samples, thanks to the ImageStreamX’s ability to image large numbers of cells from each sample.

Acquired data can be analyzed on the ImageStreamX workstation. This workstation is for temporary storage and all data should be moved to another location after analysis.

For more details on sample preparation, click here.

For instrument layout and fluorochromes choices, click here.

For an overview of the principles of IFC and selected applications click here.

The instrument is located in Pinn Hall Room 2011 (434-243-2711). See Scheduling for information on scheduling time on this instrument.