27. Flow Cytometry and Lymphocyte Quantitation Flashcards
What cell property is forward light scatter proportional to?
Cell Size
Which cells in the peripheral blood have the greatest forward scatter? the smallest forward scatter?
Greatest FSC = Monos
Smallest FSC = RBC, plts, debris
Mono > PMN > Lymphs > RBC, plts, debris
What cell property is side scatter proportional to?
Cell Complexity (granularity and membrane integrity)
Which cells in the peripheral blood have the greatest side scatter? the smallest side scatter?
Greatest SSC = PMN
Smallest SSC = RBC, plts, debris
PMN > Mono > Lymphs > RBC, plts, debris
Indicate the peripheral blood populations depicted in the attached histogram.
X-axis = FSC
Y-axis = SSC
A. RBC, plts, debris
B. Lymphocytes
C. Monocytes
D. Granulocytes
Refer to the attached histograms. In flow histogram 1, indicate what cells are present in quadrants 1 and 4. In flow histogram 2, indicate what cells are present in quadrant 2.
Histogram #1 = CD3 on x-axis, CD19 on y-axis
Histogram #2 = CD3 on x-axis, CD4 on y-axis
Histogram 1:
Q1 = T cells
Q4 = B cells
Histogram 2:
Q2 = t helper and t regulatory cells
Suppose a patient had 50% CD 19 - positive cells in his peripheral blood. What would that tell you about the patient’s condition?
Elevated level of B cells
- CD 19 = B cells
- RR for B cells = 5-25%
Suppose a patient had 25% CD 3 - positive cells in his peripheral blood. What would that tell you about the patient’s condition?
Decreased level of T cells
- CD 3 = all T cells
- RR for T cells = 60-85%
In order to quantitate the number of B cells and NK cells in a patient’s peripheral blood by direct immunofluorescence, what monoclonal antibodies would the patient’s white blood cells be incubated with? What fluorescent dyes would be attached to these antibodies?
B cells = CD 19 or CD 20
NK cells = CD 16 or CD 56
Fluorescent dyes = FITC (green), PE (orange), PerCP (red), or ACP (red)
