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