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Hematocrit: Method of Measurement
Hematocrit: Method of Measurement
Hematocrit (HCT) and packed cell volume (PCV) are used to measure red blood cell mass.
An increase in red blood cell mass is equivalent to polycythemia and a decrease indicates
an anemia. Because HCT and PCV are affected by changes in water, the HCT and PCV
may be high due to water losses in a dehydrated animal (depending on the extent of
dehydration). This can normalize a HCT/PCV in an anemic animal or may increase the
HCT/PCV above the upper reference limit in a non-anemic animal, resulting in a relative
polycythemia.
Method of measurement
PCV tube
Although used synonymously, HCT and PCV actually represent different ways of measuring
the proportion of blood composed of red blood cells.
Hematocrit: This is actually a calculated value obtained from modern automated
hematology analyzers. It is the product of the mean cell volume (MCV) and the red
blood cell (RBC) count, both of which are directly measured by the analyzer. Therefore,
if there are any inaccuracies in measurement of the MCV or RBC count, the HCT will
reflect those inaccuracies. The formula used to calculate the HCT is as follows:
HCT = (MCV x RBC count)÷10
Thus, anything that falsely increases or decreases the MCV (e.g. storage of RBC may
result in RBC swelling with an increased MCV, thus falsely increasing the HCT) or RBC
count (e.g. hemolysis will falsely decrease) will affect the HCT, but not necessarily, the
PCV.
Packed cell volume (PCV): This is a directly measured value obtained from
centrifuging blood in a microhematocrit tube in a microhematocrit centrifuge. The PCV is
measured as the height of the red cell column in a microhematocrit tube after
centrifugation (see image at right). It is the quickest and most readily available measure
of the red blood cell component of blood. Unlike the HCT, this measurement is affected
by plasma trapping and how the red blood cells pack within the column. Red blood cell
packing is species-dependent – it takes longer for ruminant RBCs to pack compared to
dogs, cats and horses, Therefore, the microhematocrit tubes are spun for 10 minutes in
ruminants versus 3 minutes in other species. Examination of the “crit tube” can also
provide subjective information about the color and clarity of the plasma (icterus,
hemolysis, lipemia), and the size of the “buffy coat” (which contains WBC and platelets).
Additionally, one can score and break the tube as desired to remove the plasma for
refractometric protein estimation, or to extrude the buffy coat for smear-making. The
“buffy coat smear” has the advantage of providing a concentrated preparation of
nucleated cells, which can be useful if looking for low-incidence infectious agents
(e.g., Anaplasma organisms in leukocytes).
Units of measurement
Both are expressed as % of the blood (SI units are L/L). The conversion formula to SI units
is as follows
% ÷ 100 = L/L
Sample considerations
Sample type
Whole blood, body cavity fluids (PCV only)
Anticoagulant
EDTA is the preferred anticoagulant. Although citrate can be used, the volume of citrate in
the tube (10% of the collection volume) will dilute the PCV or HCT accordingly. Heparinized
whole blood can also be used.
Stability
HCT and PCV are optimally stable for 24 hours at 4°C. After this time, RBC tend to swell
which increases the MCV, falsely increasing the HCT and PCV potentially (RBC do not
pack as well). Also red blood cells start to hemolyze with storage, resulting in false
decreases in both HCT and PCV.
Interferences
Lipemia, icterus: No effect.
Hemolysis: Will falsely decrease the HCT and PCV. In an animal with true intravascular
hemolysis versus in vitro hemolysis, the PCV or HCT is a better indicator of the oxygen
carrying capacity of blood than the hemoglobin.
Test interpretation
Increased values (polycythemia, erythrocytosis)
Artifact: Insufficient centrifugation speed may falsely increase the PCV if the RBC do
not pack properly. With false increases in MCV (e.g. storage of blood), the HCT will be
falsely increased, but usually the results do not exceed the upper reference limit
(however a mildly anemic animal may no longer appear anemic).
Pathophysiologic
o Relative change to blood water: Dehydration, splenic contraction secondary to
epinephrine (horses).
o Absolute increase in RBC mass: Stimulated by erythropoietin
(secondary polycythemia) or erythropoietin-independent (primary polycythemia)
Decreased values
Artifact: Hemolysis of RBC due to sample collection or storage. In this setting, the
measured hemoglobin is the most accurate measure of the animal’s oxygen carrying
capacity and a HCT can be estimated by multiplying the hemoglobin x 3 (RBC contain
1/3 hemoglobin in most species). With false decreases in MCV (e.g. excess EDTA), the
HCT will be falsely decreased, but usually the results are not below the lower reference
limit.
Pathophysiologic:
o Relative change to blood water: Over-dilution with fluids, splenic relaxation
(anesthetic agents, tranquilizers).
o Absolute decrease in RBC mass: Indicates a true anemia, due to hemorrhage,
hemolysis (intravascular, extravascular) or decreased production. Multiple
mechanisms may be operative.
Nilai hematokrit adalah volume semua eritrosit dalam 100 mL darah dan disebut
dengan persen (%) dari volume darah tersebut. Biasanya nilai hematokrit ini ditentukan dengan
menggunakan darah vena atau darah kapiler. Ada 2 (dua) cara dalam menentukan nilai hematokrit,
yaitu :
–
A. MAKROMETODE (MENURUT WINTROBE)
1. Isilah tabung Wintrobe dengan darah antikoagulan oxalat, heparin, atau EDTA sampai garis
tanda 100 di atas.
2. Masukkan tabung tersebut ke dalam sentrifuge (pemusing) yang cukup besar, pusinglah
selama 30 menit dengan kecepatan 3000 rpm.
3. Bacalah hasilnya denan memperhatikan :
Warna plasma di atas : warna kuning itu dapat dibandingkan dengan larutan kaliumbicarbonat
dan intensitasnya disebut dengan satuan. Satu satuan sesuai dengan warna kaliumbicarbonat 1
: 10000.
Tebalnya lapisan putih di atas sel-sel merah yang tersusun dari leukosit dan trombosit (buffy
coat)
Volume sel-sel darah merah
–
B. MIKROMETODE
1. Isilah tabung mikrokapiler yang khusus dibuat untuk penetapan nilai hematokrit mikrometode
dengan darah.
2. Tutuplah salah satu ujungnya dengan membakarnya dengan nyala api atau dapat juga
digunakan bahan penutup khusus.
3. Masukkanlah tabung mikrokapiler tersebut kedalam sentrifuge khusus yang dapat mencapai
kecepatan besar, yaitu lebih dari 16000 rpm (sentrifuge mikrohematokrit)
4. Pusinglah selama 3-5 menit
5. Kemudian nilai hematokrit dengan menggunkan grafik atau alat khusus
Results from your hematocrit test are reported as the percentage of blood
cells that are red blood cells. Normal ranges vary substantially with race, age
and sex. The definition of normal red-blood cell percentage also varies from
one medical practice to another.
Your hematocrit test provides just one piece of information about your health.
Talk to your doctor about what your hematocrit test result means in light of the
symptoms you're experiencing and the results of other diagnostic tests.
A number of factors can affect the outcome of a hematocrit test and yield
inaccurate or misleading results, including:
NOTE: This is the Consumer Version. DOCTORS: Click here for the
Professional Version
Biology of Blood
Overview of Blood
Components of Blood
Formation of Blood Cells
Effects of Aging on the Blood
The main components of blood include
Plasma
Red blood cells
White blood cells
Platelets
Plasma
Plasma is the liquid component of blood, in which the red blood cells, white blood cells, and
platelets are suspended. It constitutes more than half of the blood's volume and consists
mostly of water that contains dissolved salts (electrolytes) and proteins. The major protein
in plasma is albumin. Albumin helps keep fluid from leaking out of blood vessels and into
tissues, and albumin binds to and carries substances such as hormones and certain drugs.
Other proteins in plasma include antibodies (immunoglobulins—see Acquired Immunity :
Antibodies), which actively defend the body against viruses, bacteria, fungi, and cancer cells,
and clotting factors, which control bleeding (see How Blood Clots).
Plasma has other functions. It acts as a reservoir that can either replenish insufficient water
or absorb excess water from tissues. When body tissues need additional liquid, water from
plasma is the first resource to meet that need. Plasma also prevents blood vessels from
collapsing and clogging and helps maintain blood pressure and circulation throughout the
body simply by filling blood vessels and flowing through them continuously. Plasma
circulation also plays a role in regulating body temperature by carrying heat generated in
core body tissues through areas that lose heat more readily, such as the arms, legs, and
head.
Neutrophils, the most numerous type, help protect the body against infections by killing
and ingesting bacteria and fungi and by ingesting foreign debris.
Lymphocytes consist of three main types: T cells (T lymphocytes) and natural killer cells,
which both help protect against viral infections and can detect and destroy some cancer
cells, and B cells (B lymphocytes), which develop into cells that produce antibodies.
Monocytes ingest dead or damaged cells and help defend against many infectious
organisms.
Eosinophils kill parasites, destroy cancer cells, and are involved in allergic responses.
Some white blood cells flow smoothly through the bloodstream, but many adhere to blood
vessel walls or even penetrate the vessel walls to enter other tissues. When white blood cells
reach the site of an infection or other problem, they release substances that attract more
white blood cells. The white blood cells function like an army, dispersed throughout the
body but ready at a moment's notice to gather and fight off an invading organism. White
blood cells accomplish this by engulfing and digesting organisms and by producing
antibodies that attach to organisms so that they can be more easily destroyed (see White
blood cells).
When the number of white blood cells is too low (leukopenia), infections are more likely to
occur. A higher than normal number of white blood cells (leukocytosis) may not directly
cause symptoms, but the high number of cells can be an indication of a disease such as an
infection or leukemia.
Platelets
Platelets (also called thrombocytes) are cell-like particles that are smaller than red or white
blood cells. Platelets are fewer in number than red blood cells, with a ratio of about 1 platelet
to every 20 red blood cells. Platelets help in the clotting process by gathering at a bleeding
site and clumping together to form a plug that helps seal the blood vessel. At the same time,
they release substances that help promote further clotting. When the number of platelets is
too low (thrombocytopenia), bruising and abnormal bleeding become more likely. When the
number of platelets is too high (thrombocythemia), blood may clot excessively, causing a
stroke or heart attack.