Amnis ImageStream Mark-II Quick Start Manuals

ImageStream Mark-II Operation
Quick Start Guide

To begin normal operation of ISX- MkII:

1. Power up the system and launch the
2. Check to be sure the buffer containers are full and the waste tank is empty.
3. Select Startup
4. In the Calibrations view, press
when all tests pass the ASSIST button turns green and the system is ready to run for the day.
5. Select file and load
6. Press Load and, load an aliquot of a sample with each fluorochrome present.
7. In the Illumination section, turn on the appropriate
experiment.
8. Adjust the laser power to maximize brightness and
9. Create dot plots and
10. Set the acquisition parameters
population(s) to collect.
11. Chose file format, either .rif (IDEAS), .fcs (FACS software) or both in the file drop down.
12. Collect compensation
channels enabled) or by using the compensation wizard and following the prompts.
13. Continue collecting all experiment files using consistent instrument settings (In general brightfield
will be in channels 1 and 9, SSC ~40mw in channel 6, and the cells to collect R1 using brightfield
area vs. aspect ratio to identify single cells).
14. Save an experiment
15. Shut the system off by pressing the
The ImageStream MkII is a class 1 laser instrument.
ISX
and the instrument will flush the system and load sheath in ~12min.
Start all calibrations and
default template or an experiment template from the File menu.
regions to identify the cells to collect, or collect all events.
including file name, destination folder, number of events and
control files with manual settings (brightfield and 785 OFF and all
template by selecting Save Template from the File menu.
Shutdown
application.
tests. Calibration takes ~10min and
lasers for each fluorochrome in the
prevent saturation.
button. The system will sterilize itself in ~40min.