Single-cell Flow Cytometer

Amnis ImageStreamX Mk II

The Amnis ImageStreamX Mk II is a multispectral imaging flow cytometer equipped with a CCD camera capable of capturing up to six simultaneous images of each cell or object in brightfield, scatter, and multiple fluorescent channels at speeds of up to 5,000 objects per second.
Suspended cells are hydrodynamically focused into a flow cell, where they are illuminated by both a brightfield light source and lasers. High numerical aperture (NA) objectives collect fluorescence emission, along with light scattered and transmitted by the cells. These objectives then intersect a spectral decomposition element, which focuses different spectral bands onto different physical positions of the CCD camera. By collecting a large number of digital images per sample and providing a numerical representation of image-based features, the ImageStreamX Mk II combines the information of standard single-cell microscopy with the statistical significance of large samples in standard flow cytometry.
While it can be used to measure fluorescence intensity like conventional flow cytometers, the most beneficial applications of the ImageStreamX Mk II leverage its imaging capabilities to detect and quantify signal distributions within cells or even between cells in cellular conjugates.
The system also features dedicated software for spectral compensation, image analysis, and statistical analysis of acquired images with predefined guided procedures for apoptosis, cell cycle, co-localization, internalization, shape change, spot counting, and more.
The instrument is equipped with:
  • An automatic sampling system for multi-well plates, with volumes ranging from 20 to 200 μL.
  • High-speed imaging (imaging rate of 5,000 cells/sec).
  • Multi-magnification systems:
    • 60X, NA 0.90
    • 40X, NA 0.75
    • 20X, NA 0.50
  • 6 imaging channels (1 brightfield channel + 5 channels for fluorescence detection).
  • 6 LASER sources:
    • λ=488 nm, P=400 mW
    • λ=375 nm, P=70 mW
    • λ=405 nm, P=120 mW
    • λ=642 nm, P=150 mW
    • λ=561 nm, P=200 mW
    • λ=785 nm (dedicated to side scatter)

📍 The ImageStreamX Mk II imaging flow cytometer is located at the Nanobioelectronics, Nanobiotechnology, and Nanomedicine Laboratories of the Department of Mathematics and Physics of the University of Salento.

Go Back