ThermoFisher Scientific invitrogen iSort Series, Manual

Page: 38 / 140

38 iSort
™
Automated Cell Sorter User Guide
Analyze or Sort sample
Overview As the cells or particles in the sample pass through the optical detection area, they
are interrogated by a 488-nm laser, which generates scattered light and fluorescence
signals that are displayed in the fluorescence scatter plot on the screen.
Scatter plot on Define Regions screen Scatter plot on Sort screen
•
Each data point (i.e., event) displayed on the fluorescence scatter plot represents
a single cell or particle measured.
•
The position of an event in the plot is determined by the light scatter (y-axis)
and fluorescence (x-axis) measurements. The light scatter value reflects the cell
size and granularity, while the fluorescence value represents the expression
level of the GFP reporter gene or the brightness of the label or dye.
•
The accumulation of events in the scatter plot
allows you to discern specific clusters that can
represent discrete populations in your sample.
•
The number of events registered for specific
populations in the sample are indicated by
different colors in a density color gradient, where
light blue, green, yellow, and red hot spots
indicate increasing number of the events (blue
represents the lowest and red represents the
highest density).
Note:
The presence and position of discrete clusters in the plot are entirely
sample dependent.
•
You can draw gates around the target populations to define them as Region 1,
Region 2, or debris for Analysis or Sort.
•
When sorting (page 41), the target population defined by Region 1 is sorted into
the collection tube placed in Position 1 (left) in the collection tube holder, while
the target population defined by Region 2 is sorted into the collection tube
placed in Position 2 (right).