Professional Documents
Culture Documents
HISTOGRAM GENERATION
AND INTERPRETATION
Dr. N. Bajaj
What is a histogram?
Title
Horizontal or X-axis
Bars:
Vertical or Y-axis
Interpretation of Histogram
Most of the data were on target, Although some data were on target,
with very little variation from it many others were dispersed away from
the target
Even when most of the data The data were off target and widely
were close together, dispersed
they were located off the
target by a significant amount
Histogram in haematology
Histogram Characteristics
Histograms provide information about erythrocyte, leukocyte and platelet frequency and
distribution as well as depict the presence of subpopulations.
The passage of each individual cell momentarily increases the impedance (resistance)
of the electrical path between two submerged electrodes that are located on each side
of the aperture.
The number of pulses generated during a specific period of time is proportional to the
number of particles or cells. the amplitude (magnitude) of the electrical pulse produced
indicates the cell's volume.
Raw data from cell analyzer
raw data is generated and the analyzer's computer classifies the raw data
• The volume histogram reflects the size of erythrocytes or any other particle in the
erythrocyte size range. The instrument counts the cells as sizes between 25 fL and 250
fL.
• RBC histogram is a symmetrical bell-shaped curve.
• The area of the peak is used to calculate the MCV and RDW. This area represents 60 fL
to 125 fL.
• If the RBCs are larger than normal (macrocytic), the curve will shift toward the right.
• If the RBCs are smaller than normal (microcytic), the curve will shift to the left.
• The extension of the lower end of the scale allows for the detection of erythrocyte
fragments, leukocyte fragments and large platelets.
• If the histogram curve is bimodal (Camel humps), then there are two populations of red
blood cells as might be seen when a patient received a blood transfusion.
Other conditions that will cause a bimodal distribution curve are cold agglutinin
disease, hemolytic anemia with presence of schistocytes, or anemias with different
size cell populations.
If the leukocyte count is significantly elevated, the erythrocyte histogram will be
affected as in the case of CLL.
The mean corpuscular volume (MCV) is calculated from the area under the peak.
The red blood cell distribution width (RDW) is also calculated from the data used
to calculate the MCV.
Interpretation of the RBC histogram, along with the numerical values of RBC
count, Hgb, Hct, MCH, MCHC and RDW can significantly assist in the diagnosis
of various RBC disorders avoiding many expensive and invasive investigations
Possible causes:
• Platelet clumps
• RBC fragments
• Giant platelets
• Microerythrocytes
Possible cause:
• It R RBC agglutination (cold agglutinins)
causes the RU flag and it disappears
when these samples are incubated at
room temperature.
• RU flags may also be seen when small ss
lymphoid cells are present in very high
number like in CLL.
Possible causes:
• Same as RU or RL
Possible causes:
Iron deficiency in recovery (post-
therapy)
Post-transfusion
Infect or tumor anemia (visceral iron
deficiency)
Extreme leukocytosis
THE RBC HISTOGRAM
All values are normal except anemia and the peripheral blood smear and histogram
resembles the seen in normal subjects.
Almost similar picture may be seen in:
•Anemia of acute blood loss
•Hypothyroidism
•CRF
•Radiation or drug suppressed marrow
•Chronic Liver Disease
•Anemia is usually caused by multiple mechanisms, e.g. hypoproliferative (failure of
erythropoiesis), decreased RBC survival, iron deficiency of chronic inflammation,
impaired erythropoietin, etc.
•Anemia is normocvtic. normochromic in most of the cases.
•RDW and indices are usually normal.
•Anemia may be hypochromic and or microcvtic in 25 to 3O° of the cases.
•Moderate anisocytosis and slight poikilocytosis may be seen.
•Low reticulome count.
•Polvchromatophilia. and nucleated RBCs are absent.
•Serum ferritin is increased or normal in contrast to iron deficiency.
•Free erythrocyte protoporphyrin is increased.
In case of advanced megaloblastic anemia with severe folate deficiency
•RBC count is low, MCV is high, and RDW is increased.
•Histogram is widened and shifted to right distinctly, indicating macrocytosis. Large
hypersegmented neutrophils with low Hb, increased MCV, raised RDW with presence
of macro ovalocytes are considered diagnostic.
IN CASE OF EARLY MEGALOBLASTIC ANEMIA
The MCV is still normal RBC count and Hb slightly reduced.
RDW is distinctly increased.
Histogram is widened on right side, indicating appearance of macrocytic
population, although not yet enough to increase MCV.
Raised RDW with changes in histogram may alert the clinician for the
possibility of early megaloblastic anemia even before apparent anemia.
Normocytic recovery (circle and arrow)
A small peak of cells in the normal range.
RDW is higher than untreated megaloblastic anemia due to two cell population
contributing to the heterogeneity.
Microcytic recovery (arrow)
Two cell populations are clearly seen in this histogram—Old macrocytes and newly
produced or surfaced up microcytes.
Concomitant iron deficiency has been unmasked.
RDW is markedly increased due to increased heterogeneity.
MCV is normal only because it reflects the average of two abnormal populations.
Double populations of red cells may exist in:
Prrsence of microcytic and Macrocytic cells in same sample may result in a normal MCV and may
mislead if only observed numerically.
Hb and Hct levels are decreased.
RBC count decreased, destruction by the mechanism of immune adherence by the
patient produced antibodies.
At maximum hemolysis reticulo-cytosis is profound.
MCV is increased due to reticulo-cytosis.
RDW also increased to make a macrocytic heterogeneous anemia.
Key diagnostic feature of various common haematological conditions
RBC plot on
Condition Hb MCV MCH MC HC RDW Cystogram
Normal Normal Normal Normal Normal Normal In normocytic
normochromic zone
Iron deficiency Low Low Low Normal or low High Microcytic hypochromic
anemia zone (triangular spread)
Beta Normal Low Low Normal or low Normal or Narrow clustering in the
thalassemia or low near normal microcytic hypochromic
trait (minor) zone (comma-shaped)
Beta Very low Low Low Low Very high Widespread in the
thalassemia micro-cytic hypochromic
(major) zone (simulates an
exaggerated version of
the cystogram seen in
iron deficiency)