Abbott- Hematology learning guide

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- The average adult circulation contains 5 liters of blood
(roughly 5.28 quarts).
Blood completes the entire systemic circuit from left heart
through the body to right heart in 90 seconds.
Every cubic millimeter of blood contains 5 million RBCs.
RBCs survive about 4 months; neutrophils survive about 6
hours

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-Pada yahun 1800 Paul Erlich memperkenalkan istilah eritroblas utk menyatakan prekursor eritrosit berinti
-Pada tahun 1949 ASCP (American society clinical pathologist) menyamakan nama menjadi rubri (merah) blast
-Red cells which have a blue or lilac tinge are said to show polychromasia (many colours).
- Polychromatic cells are young cells, newly released from the bone marrow.
-They have not yet been remodelled to the disc shape of a mature erythrocyte and therefore lack central pallor
-They are also generally larger than more mature cells and in this case may be described as polychromatic macrocytes
-Young red cells can also be detected with a special stain of live (unfixed) cells called a supravital stain
-Young cells detected in this way are called reticulocytes because the supravital staining causes a network or
reticulum to be deposited.

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-- Platelets form by fragmentation of megakaryocyte membrane exvaginations termed proplatelets
-Each megakaryocyte is estimated to give rise to 1000 to 3000 platelets (before the residual nuclear material is
engulfed and eliminated by marrow macrophages)

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-PLT with a size from 8-12 fl are counted between 2 and 30 fl.
-A fixed discriminator at 12 fl .
-The Erythrocytes do have a size from 80-100 fl and are measured between 25 and 250 fl

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 SLS : Cyanide-free and non-toxic (an integral part of soaps).

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10 ribu rpm 2-4 menit, dgn tabung kapiler, nama lain PCV (packed cell volume)

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In case of RBC fragments and extreme microcytes there is no clear separation in volume between platelets
and erythrocytes. Due to high numbers of RBC the platelet result might be false high and should be
checked with alternative methods (chamber count or Fonio). In most cases the RBC result is only slightly
interfered

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RBC agglutination might cause a low incorrect RBC result and effect also the parameters HCT, MCV, MCH
and MCHC. In case of cold agglutinates warm up the sample to 37C. (MCHC should drop back to normal
value if the problem is solved.) Histogram

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Often an extreme anisocytosis is found. In case of anisocytosis the RBC result is not affected. Seldom can
a MP occur in samples with extreme high numbers of leukocytes and the RBC results may be incorrect
high. Therefore all RBC parameters should be controlled.

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The flag DW is shown in case of abnormal histogram curve. The overall height of the curve represents
100%. The width is calculated on the 20% line of the curve. If the histogram curve does not match the 20%
line neither the lower nor upper discriminator the flag DW is generated for RDW-SD or RDW-CV and
these results can not be calculated.

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 Primary disorder = ggn adhesi (pada BT) dan agregasi

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 - This platelet value can be used if the MCV shows a
standard value. In this case, a chamber count would be
statistically much more imprecise

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 -Fragmentocytes slip into the platelet population and do
not allow any clear separation between red blood cells
and platelets.
-In any event, this platelet value is questionable and
requires verification.

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Case Answer: . The Effect of Giant Platelet
Pic1.This is a sample with the appearance of large platelets sized almost equal to erythrocytes. The abnormally wide
distribution on the PLT histogram suggests the appearance of large platelets. The distribution curve intersects the
discrimination line at a low point ( ), which shows the platelet count has been measured correctly.

Pic 2.This is a sample with the appearance of large platelets. Although wide distribution on the PLT histogram
suggests the appearance of large platelets, the distribution curve intersects the discrimination line at a high point ( ).
This result needs to be confirmed by other methods i.e. Fonio method, Rees-Ecker method or
Brecher-Cronkite method.

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Answer: Cold Agglutinin
Sample needs to be warmed up at 370C for 30-60 min and analyze immediately. Seen in cases of multiple
myeloma.

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 CBC and DIFF (28 parameters)
Flexible configurations: from benchtop
system to independent workplace
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Reflex sampler for increased

walk-away time
Integrated reflex rule set
Optional wagons for each system
for best organisation of the workplace
storage for reagents and pneumatic unit

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Signal is changed from SSC to SFL.

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Signal is changed from SSC to SFL.

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Calculation of actual count = 10,000 * 58.2/61.05
Undiluted sample volume = 0.9532

In LWBC, the actual cell count by analyser is 3 times more.

Actual cell get counted =

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Beneficial for high workload laboratories with large number of abnormal samples
Reflex testing combined with special rule set,after initial analysis

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The white blood cell count is markedly high with a high monocyte fraction of 59.8%. While blasts are the main
components in the manual count, the WDF scattergram shows a poor separation. It seems that these cells are
plotted near the area where normal monocytes appear (). In the WPC scattergram, most of the blasts are found in
the area; some are detected as abnormal cells in the WPC and WPC (SSC-FSC) scattergrams, with the Blasts?
flag displayed.

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Specific staining of platelets only .

Fragmentation is seen in both Beta thal Sickle cell A.

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Calculation of actual count = 200,000 * 18.21/203.8
Undiluted sample volume = 0.08935

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Fragmented erythrocytes increase, platelet decrease

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Value of PLT-F is a precise platelet count that can be used reliably to decide
platelet transfusion

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Interference PLT-I sample; -Thalassemia, PLT-I is 514, PLT-F is 140.8, CD61/41 is 152.2
XE: Of course the PLT-I result is flagged as unreliable and switched to PLT-O result!
Blood film confirmed
See animation for second example

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Interference PLT-O sample; APL Chemo, PLT-O is 181, PLT-F is 24, CD61/41 is 24. PLT-I was already correct and
no Flag PLT reflex appears
XE: The PLT-O result is flagged as unreliable and switched to PLT-I results!
Blood film confirmed
Apoptotic WBC or cytoplasm fragments (often present during chemotherapy) may be stained by the polymethine dye

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-The determination of the reticulocyte count and classification (maturation index:
LFR, MFR, HFR) give information on the activity level of erythropoiesis.
-It is usual to give the reticulocyte value in per cent or per mill of the red blood cell
count.
-Mature RBCs and Reticulocytes (RET) are fractionated by their RNA content
difference.
-Reticulocytes and WBCs are fractionated by their DNA and RNA content differences

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-The interpretation of the reticulocyte count is problematic in severe anaemias.
-A moderately increased relative reticulocyte count in severe anaemia does not
indicate a sufficiently strong regeneration of erythropoiesis, but merely indicates a
shortened life span of the red blood cells.
-It is preferable to give the absolute reticulocyte concentration in reticulocytes/L, as
this provides a direct measurement of erythropoietic performance.
-For example, a value of 20 reticulocytes is considered increased. However, in
severe anaemia with 2 million red blood cells, 20 reticulocytes merely represent
40,000 reticulocytes/L, a value in the lowest reference range

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 -Corrective measures can be carried out via the patient's haematocrit by reference to a normal haematocrit of 0.45
[L/L].
-To be able to evaluate whether the regenerative power of erythropoiesis is adequate, the reticulocyte count (RET) has
to be corrected depending on the haematocrit

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-The maturation time of reticulocytes in the bone marrow is proportional to the haematocrit, i.e. it decreases with the
haematocrit, and the maturation time in the blood increases
-To give an indication of the efficiency of the bone marrow, the reticulocyte count is corrected by this haematocrit-
dependent factor

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-The RPI is an index which helps to evaluate erythropoiesis efficiency and thus the productivity of the bone marrow
-RPI is an indicator of the rate of RBC production and increases in response to anemia.
-An RPI value >3 generally indicates an adequate bone marrow response to anemia, whereas an RPI value <2
represents an inadequate response

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Stiene Martin, Clinical hematology, page 116

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-IRF percentage increases after only a few hours, the reticulocyte count increases after two to three days
-Monitoring transplant succes in 80 % of cases the IRF value reaches its 5 % mark earlier than the granulocytes their
classic threshold of 0.5 x 109 granulocytes/L

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- Differentiate between actual iron deficiency and functional iron deficiency (disturbance of iron mobilisation)

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- RET-He: RET-He is calculated from
RET-Y and represents the reticulocyte Hb
equivalent in pg

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 -The RPI is an index which helps to evaluate erythropoiesis efficiency and thus the productivity of the bone marrow
-RPI is an indicator of the rate of RBC production and increases in response to anemia.
-An RPI value >3 generally indicates an adequate bone marrow response to anemia, whereas an RPI value <2
represents an inadequate response

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 -The maturation time of reticulocytes in the bone marrow is proportional to the haematocrit, i.e. it decreases with the
haematocrit, and the maturation time in the blood increases
-To give an indication of the efficiency of the bone marrow, the reticulocyte count is corrected by this haematocrit-
dependent factor

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Explanation:
In case of
platelet
aggregation,
the PLT count
is incorrect low.
Check EDTA-
incompatibility
e.g. recollect
the sample and
use sodium
citrate as
anticoagulant
to avoid clotting
caused by
EDTA. In case
of giant
platelets, the
PLT count
might be
incorrect low. In
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case of
extreme
microerythrocyt
es or RBC
fragments the
PLT count
might be
incorrect high.
The PLT result
should be
confirmed with
alternative
methods: e.g.
chamber
counting or with
Fonio method! Company Confidential Copyright 2015 PT Sysmex Indonesia
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- In case of lymphatic leukaemias the IMI channel is

usually empty (exceptions are only in very seldom cases
of acute lymphatic leukaemias known)

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- In case of lymphatic leukaemias the IMI channel is

usually empty (exceptions are only in very seldom cases
of acute lymphatic leukaemias known)

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