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The ImageStreamX (ISx) is a multiparameter imaging cytometer that combines the imaging capability of a wide-field fluorescence microscope and the high throughput, multiparameter ability of a flow cytometer.
The ISx has 12 imaging channels and can acquire up to 9 fluorescent channels from a single sample.
Features
Features
4 lasers (488nm, 561nm, 405nm and 640nm), all independantly controlled and power-tunable
12 channels including brightfield and dark field (scatter) channels
9 colour
up to 500 events per second
20x, 40x and 60x Olympus objectives
96 well plate reader for automated, unassisted acquisition
identification of cells post acquisition to enable effective data analysis
Wide array of image analysis tools and protocols
Applications
Applications
The ISx supports a wide range of applications which are detailed below. In addition, the ISx has demonstrated the ability to identify submicron particles (eg exosomes, microparticles) with resolution down to 50-100nm. As a member of the Flow team about these applcations or contact us on coreflowlab@manchester.ac.uk
Resources
Resources
Sample Preparation guide (Amnis)
Main points regarding sample preparation:
Cells can be live or fixed (acquisition can be quite slow so consider cell health if you are running live cells)
Samples should be brought in 1.5mL eppendorff tubes in a final volume of 30-100uL of PBS
Staining protocols are the same as you would perform for flow cytometry
Panel design can be a little different due to the arrangement of the lasers, so please discuss this with the facility manager prior to commencing any practical work
We advise performing titrations of antibody panels to obtain the best signal to noise and optimise population resolution
Compensation controls and fluorescence minus one (FMO) controls are advised. Please discuss this with the facility manager ahead of your experiment.
Configuration (Standard)
Configuration (Standard)
The configuration is detailed in the table below and customizable with regards to channels and laser so please feel free to discuss this.
More to come...
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