Beckman Coulter COULTER LH 750 System, Справочник, Страница: 45 / 120

PN 4277248DD
3-13
OPERATION PRINCIPLES
DATAPLOT DEVELOPMENT
3
converted to absorbance, the reagent-blank signal level provides a reference to which the
sample signal is compared.
3.6 DATAPLOT DEVELOPMENT
At the Workstation, raw data is analyzed to:
r
Position each cell within a three-dimensional space
r
Plot the data on a two-dimensional DataPlot
r
Display and report results.
The Workstation performs a series of operations on the stored digital raw values received from the
flow cell to identify memberships (subpopulations) and calculate the frequency of cells within
each membership group. It also produces the DataPlot displays for visual representation of the
WBC Diff and Reticulocyte/RBC membership and density.
In the LH 750, the Beckman Coulter AccuGate algorithm uses adaptive contouring methods
designed for finding optimal separation between overlapping clusters of data. The AccuGate
algorithm provides a statistical analysis tool that discovers the overlapping populations using
nonlinear separation techniques. The newly developed adaptive gating techniques use
multidimensional data to distinguish the presence of even the faintest subpopulation. The
AccuGate algorithm can adapt to unusual population shifts and overlaps. It can define highly
irregular separation, and signal the need for further analysis. It can then make a subsequent
analysis of the identified regions and correct deficiencies in separation.
In the DataPlots, different colors represent different memberships (types of cells). Shades of colors
represent density (concentration): dark colors for low density, bright colors for high density.
Two-Dimensional (2D) DataPlots
The 2D WBC Diff DataPlot shows the five memberships: lymphocytes (LY), monocytes (MO),
neutrophils (NE), eosinophils (EO) and basophils (BA), plus the non-white cell populations.
Cell volume (VOL), determined by the low-frequency impedance measurement, is plotted on
the Y-axis; rotated light scatter (RLS) is plotted on the X-axis.
The Reticulocyte DataPlot shows mature red cells and Reticulocytes. Cell volume is plotted
on the Y-axis, and linear light scatter (LLS) is plotted on the X-axis.
Three-Dimensional (3D) DataPlots
The 3D DataPlot view classifies by density, light scatter and opacity. The axes are color coded.
DIFF: Lymphocytes Blue
Neutrophils Purple
Eosinophils Orange
Monocytes Green
Basophils White
Non-white debris Red
RETIC: RBCs Red
Reticulocytes Blue
Platelets Green
WBCs Purple