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Methods
Reticulocyte Method
Cytochemical Reactions
The reticulocyte cytochemical reactions consist of two steps:
Step 1
RBCs and platelets are isovolumetrically sphered using ADVIA
2120/2120i autoRETIC reagent.
Step 2
Reticulocytes are differentially stained based on their RNA content.
ADVIA 2120/2120i autoRETIC
NOTE:
For more detailed information on the contents of ADVIA 2120/2120i
autoRETIC reagent, please see Chapter 8.
The ADVIA 2120/2120i autoRETIC reagent contains a zwitterionic detergent
(surfactant) that isovolumetrically spheres the red cells. It also contains a cationic
dye, Oxazine 750, that stains cells according to their RNA content.
Measurement
A constant volume of the cell suspension from the retic reaction chamber passes
through the flowcell where the low-angle light scatter (2° to 3°), the high-angle
light scatter (5° to 15°), and the absorption signatures for each cell are measured.
ADVIA
2120/2120i SHEATH/RINSE encases the sample stream as the two
fluids pass through the flowcell.
The low-angle and high-angle light scatter signatures are proportional to cell size
and hemoglobin concentration. Light absorption is proportional to RNA content.
The stained reticulocytes will absorb more light than the mature RBCs.
The two light-scatter signals and the absorption signal are detected, electronically
amplified, and split into six signals.
One of the low-angle light scatter signals is amplified 28 times.
One of the high-angle light scatter signals is amplified 12 times.
One of the absorption signals is amplified 33 times.
After processing, this information is available:
RETIC Rate
histogram displays the arrival rate of cells in the retic channel.
RETIC Abs Flatness
histogram displays the mean absorption at 200-millisecond
intervals.
RETIC Abs
histogram displays the overlaid distribution of mature RBCs and
reticulocytes by their absorption values.
RETIC Volume
histogram shows the overlaid distribution of mature RBCs and
reticulocytes by their cell volume independent of hemoglobin concentration.
RETIC HC
histogram shows the overlaid distribution of mature RBCs and
reticulocytes by hemoglobin concentration independent of cell volume.
Summary of Contents for ADVIA 2120
Page 1: ...ADVIA 2120 2120i Hematology Systems Operator s Guide 067D0157 01 Rev 201 0 ...
Page 30: ...1 26 Welcome to the ADVIA 2120 2120i Hematology System ...
Page 46: ...3 12 Daily Routine ...
Page 171: ...7 2 Status Line messages T 93 U 94 V 96 W 98 X 99 Y 99 Z 99 ...
Page 269: ...7 100 Status Line messages ...
Page 341: ...8 72 Methods ...