CyFlow
®
Cube Instrument Operating Manual
Rev018_2012-01-27
10/36
G
M
L
Histogram and dotplot in immunology
Lysed blood 2D dotplot
Histogram and dotplot of immunological staining
Getting Started
M
L
G
Pos.
B
a
c
k
g
ro
u
n
d
Neg.
Slightly pos.
Lysed blood is represented in a dot plot
presenting the FSC in X axis and SSC in Y axis.
Both axes are in a linear scale.
Three distinct groups are visible; they represent
the granulocytes (G), monocytes (M) and
lymphocytes (L).
This histogram represents the spectrum of the
cells presented in the previous dot plot stained
with
antibodies
anti-CD16
conjugated
to
PhycoErytrin (PE). The X axis displays the
fluorescence in a 4-dec logarithmic scale and the
Y axis displays the number of events in a linear
scale.
This dot plot presents the cells fluorescence in X
on a logarithmic scale (CD45 FITC) versus SSC
in Y on a linear scale.
This data display allows an easier interpretation of
2 parametric data compared to the histogram.
Concluded from this example, the lymphocytes
are strongly stained with the anti-CD45 antibody,
the monocytes are slightly stained and the
population of granulocytes is negative.
This example is a simplification of an immune-
staining analysis by flow cytometry. It is
directed to new users, and aims to link the
histogram to the dotplot representation. To
complete the analysis, gating and statistics
are required.
