ABSTRACT
BACKGROUND: To achieve consistency in poikilocytes grading in peripheral blood cell examinations, we made an image-based differential count (IDC) software to measure the degree of abnormalities in individual red blood cells (RBCs) and relative fractions of poikilocytes. METHODS: Thirty peripheral blood samples were analyzed. Smear slides were examined on a microscope with charge-coupled device (CCD) camera. To verify this program, we compared the IDC results with the results of manual differential counting (MDC). Relative fractions of schistocytes, echinocytes, and elliptocytes were measured by IDC and MDC. The error rate of IDC was measured by confirming the final processed images of IDC. Correlations of IDC and MDC results were compared using linear regression analysis and the time required for each test was measured. For presentation of the mathematical decision criteria of poikilocytes, IDC algorithms for recognizing schistocytes, echinocytes, and elliptocytes were made using simple geometrical or mathematical formulas. RESULTS: The error rate of IDC was 2.8%. For analysis of 1,000 RBCs, MDC took 7.3 minutes and IDC took 2.7 minutes. Linear regression coefficients were 0.776 (P<0.001) for schistocytes, 0.895 (P<0.001) for echinocytes, and 1.001 (P<0.001) for elliptocytes. CONCLUSIONS: It was possible to define poikilocytes with geometrical or mathematical formulas using image analysis programs. The IDC program would be helpful for consistent grading of poikilocytes.
Subject(s)
Blood Cells , Erythrocytes , Linear ModelsABSTRACT
The purpose of this study is to know whether there are differences between hematology profile and morphology of erythrocytes of blood specimens which are prepared with excessive Na2EDTA anticoagulant in different concentration. This study was conducted in Faculty of Medicine Gadjah Mada University. The criteria of subject were male, age from 18 until 22 years old and healthy, ascertained from history taking and vital sign examination. Blood samples from 33 subjects were taken using vein puncture. Two millimeters blood was divided into 4 Na2EDTA-containing tubes. Before that, one drop of blood without Na2EDTA anticoagulant was used for making control blood film right after vein puncture. Each tubes contained different concentration of anticoagulant. The first tube contained Na2EDTA in standard concentration 2 mg/dl; the remaining tubes contained consecutively, 4 mg/dl, 6 mg/dl, and 8 mg/dl. Those samples were immediately examined using SYSMEX SE-9500 automatic analyzer for measuring erythrocytes hematological profile and were stained with Wright staining for morphological examination. These procedures were done before 20 minutes of vein puncture. There were significant decrease of RBC count, HGB, HCT, and MCHC and also significant increase of MCV and RDW between different concentrations of excessive Na2EDTA anticoagulant. MCH did not have significant result. Morphological examination showed significant morphological changes in the form of echinocytes and appearance of ghost cells in the sample treated with excessive Na2EDTA anticoagulant concentration. In conclusion, there are differences in hematological profile and morphology of erythrocytes among blood specimen which are prepared with excessive Na2EDTA anticoagulant in different concentration, except for MCH. Excessive Na2EDTA anticoagulant concentration will affect the blood specimen for peripheral blood examination of erythrocytes by interfering morphology and some of hematological parameters.