When erythrocytes are in solution hipotonis, osmolalitasnya fluid levels
lower than normal plasma or serum (less than 280 mOsm / kg)
Erythrocyte osmotic fragility test
(also called erythrocyte osmotic resistance) is done to measure the ability to
withstand the occurrence of erythrocyte hemolysis (destruction of red blood
cells) in asolution that hipotonis. The trick is as follows: erythrocytes were
dissolved in saline solution with various concentrations. If hemolysis occurs
at a slightly saline solution hipotonis, this state is called an increase in
erythrocyte fragility (= reduction in resistance / durability erythrocytes),
and if hemolysis occurs at a very hipotonis salinesolution, this situation
indicates decreased osmotic fragility (= increase in erythrocyte resistance).
Hemoglobin out of the cells in each
tube containing a solution of NaCl of different levels. Hb then determined
fotokolorimetrik. The results are reported in percentages (%) hemolysis.Collection of hemolysis results are plotted in a
curve as compared with normal erythrocytes data. In the state of increased
fragility, erythrocytes are usually spherical, and the curves seem to shift to
the right. While the decrease in fragility, thin and flat-shaped erythrocytes,
the curve appears shifted to the left.
Clinical Problems
DECREASE fragility: Thalassemia major
and minor (Mediterranean anemia or Cooley's anemia), anemia (iron deficiency,
folic acid deficiency, vitamin B6 deficiency, sickle cell)
hemoglobin C disease, polycythemia vera, post splenectomy, acute liver necrosis
and sub-acute, jaundice obstructive.
IMPROVEMENT fragility: hereditary
spherocytosis, transfusions (ABO and Rhesus incompatibility), autoimmune
hemolytic anemia (AIHA), hemoglobin C disease, drug toxicity or chemicals,
chronic lymphocytic leukemia, burns (thermal).
Procedure
This test is usually performed on
fresh blood samples of less than 3 hours and / atu 24-hour blood samples were
incubated at 37 ° C. Blood samples used in the form of heparin blood or blood
"defibrinated". There are no restrictions on food or beverage intake.
In this test made solution with
different concentrations of NaCl. Assessment of results with fotokolorimetri method (using photometer or spectrophotometer).
Prior to testing, provide a first
buffer stock solution of NaCl 10% made from 9 grams NaCl, 1.365 g Na2HPO4, and
0.215 grams NaH2PO4.H2O. The material is then diluted with distilled water to
100 ml. Before being used for inspection, make asolution of 1.0% NaCl principal
by dissolving 5.0 ml of saline buffer stock of 10% with distilled water to 50.0
ml. Next do the testing as follows :
1. Provide 12 pieces and then make a
dilution tube multilevel solution of NaCl concentration: 0.85%, 0.75%, 0.65%,
0.60%, 0.55%, 0.50%, 0.45%, 0, 40%, 0.35%, 0.30%, 0.20% and 0.10%, respectively
of 5.0 ml. NaCl solutions were made from the basicsolution of NaCl 1.0%.
2. Add into the canisters each 50 mL
blood sample. Mix (homogenization) by way of tossing and
turning the tube several times.
3. Inkubasikan for 30 minutes at room
temperature.
4. Mix (homogenization) again and then
worry about (centrifuges) of each tube for 5 minutes at 3000 rpm.
5. Measure the absorbance (OD) of the
supernatant at λ 540 nm with the blank tube to supernatant-1 (0.85% NaCl).
6. Calculate the% hemolysis by dividing the
absorbance (OD) of samples with the absorbance (OD) tubing to 12 multiplied by
100%.
7. Create a curve with the concentration
of NaCl as the axis (x) and% hemolysis as the ordinate (y). Compare with the
curve of normal blood controls.
Normal Value
The beginning of hemolysis on the
concentration of NaCl 0.40% - 0.45%
Complete hemolysis at a concentration
of 0.30% NaCl - 0.35%
The percentage of hemolysis in normal
circumstances are :
97-100% hemolysis in 0.30% NaCl
50-90% hemolysis in 0.40% NaCl
5-45% hemolysis in 0.45% NaCl
0% hemolysis in 0.55% NaCl
Factors Affecting Laboratory Findings
· plasma pH, temperature, glucose
concentration, and oxygen saturation in the blood
· Long-lived erythrocytes tend to have
high osmotic fragility
· Blood samples taken over 3 hours can
show increased osmotic fragility.
Source of the article
and continue reading : http://labkesehatan.blogspot.com
0 komentar:
Posting Komentar