The primitive erythroblasts in 21 cases of embryonic hearts from 4 to 9 weeks of gestation were studied with a light microscope. The nuclear diameter, the motosis, and the loss of nuclei of the primitive erythoblasts were analyzed quantitatively. The results obtained were as follows. 1. At 4 weeks of gestation, the blood cells consisted of proerythroblasts, along with basophilic polychromatophilic primitive erythroblasts. The nuclear diameter ranged from 3.20 µm to 9.20 µm, but the main range was from 4.20 µm to 6.00 µm. It was revealed that 9.50% had diameter of more than 6 µm. 2. At the fist half of the 7 week gestation when hepatic hemopoiesis developed, the blood cells consisted of basophilic, polychromatophilic, and eosinophilic erythroblasts. Cells of more than 6 µm in nuclear diameter were about 1.10% and thereafter gradually disappeared. The range of the nuclear diameters was from 2.60 µm to 7.00 µm, while a range from 3.40 µm to 5.20 µm wqs the main. The proportion of cells less than 4 µm in nuclear diameter was 39.58% and thereafter rapidly increased. 3. From the second half of 7 weeks to 9 weeks of gestation, the erythrocytes originating from hepatic hemopoiesis increasingly replaced the circulating primitive erythroblasts, which became mature during this time. The erythrocytes showed 72.88% at 9 weeks of gestation. The proportions of cells less than 4 µm in nuclear diameter in the first and second haIves of 8 weeks and 9 weeks were 52.73%, 80.02%, and 89.09%, which represented the rapid destruction of nuclei. 4. Mitosis in the primitive erythroblasts occurred principally up to the early 6th weeks, and very weakly at 8 weeks. 5. As the crown-rump length increased, the average nuclear diameter decreased very significantly (P<0.01, y=-0.2811X + 0.3171). The results suggest that distrilbution of the nuclear diameter, the maturity, the rate of nuclear loss, and the mitotic figure offer credible data for estimating embryonic age.
Basophils ; Blood Cells ; Crown-Rump Length ; Embryonic Structures* ; Eosinophils ; Erythroblasts* ; Erythrocytes ; Heart ; Humans* ; Mitosis ; Pregnancy

BACKGROUND: The National Committee for Clinical Laboratory Standards (NCCLS) recommends that the analytical variability must not exceed 25% of the biological variability in automated blood cell analysis. This study was conducted to determine whether routine automated blood cell analysis by Coulter STKS (Coulter Corp., Miami, FL, U.S.A) comforms with the NCCLS's recommendations. METHODS: Routine CBC analysis with STKS was performed on 22 healthy volunteers. The tests included calculating WBC, RBC, hemoglobin, MCV, platelet, MPV, and percentages of the granulocytes, lymphocytes, and monocytes. Blood samples were collected twice in one week interval to study the total variability. For the analytical variability, blood samples from 12 subjects were tested twice immediately after venipuncture for within-run variability, and samples from 10 subjects were tested immediately and 6 hours after venipuncture for within-day variability. The analytical variability was calculated as the sum of within-run and within-day variabilities. The biological variability was calculated by subtracting the analytical variability from total variability. The ratios of analytical and biological variabilities were calculated by dividing the analytical variability by the biological variability. RESULTS: Ratios of analytical and biological variabilities were as follows: 0.22 for WBC, 0.20 for RBC, 0.21 for hemoglobin, 0.39 for platelet, 1.98 for MPV, 0.07 for %granulocyte, 0.11 for %lymphocyte, and 1.81 for %monocyte. The ratio for MCV was not obtained because the analytical variability exceeded total variability. CONCLUSIONS: The analytical variability did not exceed 25% of the biological variability in all test categories except platelet, MPV and the percentage of monocyte. Thus, it is recommended that the analytic variability of all test categories be reduced so as to be in conformity with the NCCLS' recommendations.
Blood Cells* ; Blood Platelets ; Granulocytes ; Healthy Volunteers ; Lymphocytes ; Monocytes ; Phlebotomy

PURPOSE: To evaluate the changes of differential counts and lymphocyte subsets in cancer patients' leukocyte before and after radiotherapy. METHODS AND MATERIALS: From Dec. 1994 to May 1995, the changes of leukocyte and its subsets in 16 patients who received radiotherapy in the Dept. of Radiation Oncology of Dong-A University Hospiatal were investigated. Radiation was delivered from 2700 cGy to 6660 cGy with median dose of 5400 cGy. The results of pre- and post-radiotherapy were analyzed by paired T-test. The results of patients who received < 50 Gy and > or = 50 Gy were analyzed by wilcoxon test. RESULTS: Before and after radiotherapy, there was not any significant differences in the counts of leukocyte, granulocyte and monocyte. A remarkable decrease was noted in lymphocyte counts after radiotherapy(p=0.015). T cells, B cells and natural killer cells were also decreased in number after radiotherapy but it was not significant statistically. T helper cells and T suppressor cells were also decreased in number(p>0.05). The ratio of T helper/suppressor cell was decreased from 1.52 to 1.11 and it was significant statistically(p=0.016). The portion of T suppressor cell among all T cells was increased after radiotherapy (p=0.0195). No significant difference was observed in the analysis of leukocyte and its subsets between patients who reveived < 50 Gy and > or = 50 Gy. CONCLUSION: Radiotherapy caused remarkable decrease in lymphocyte count and its subsets. Among all lymphocyte subsets, T helper cell might be the most vulnerable to radiation, considering decreased ratio of T helper/surppressor cell count after radiotherapy.
B-Lymphocytes ; Cell Count ; Granulocytes ; Humans ; Killer Cells, Natural ; Leukocytes ; Lymphocyte Count ; Lymphocyte Subsets* ; Lymphocytes* ; Monocytes ; Radiation Oncology ; Radiotherapy* ; T-Lymphocytes ; T-Lymphocytes, Helper-Inducer

INTRODUCTION: The ABX Pentra DX 120 (Pentra DX 120, ABX Diagnostics, Montpellier, France) adopted new technologies to perform differential leukocyte and erythroblast counts. The double matrix can discriminate Large Immature Cell (LIC), Immature Granulocyte (IMG), Immature Monocyte (IMM), Immature Lymphocyte (IML), and Atypical lymphocyte (ALY) in addition to a routine 5-differential count. For erythroblast (ERB), a fluorescence method is employed. In this study, we evaluated the performance of the Pentra DX 120 in the performance of differential leukocyte and erythroblast counts. METHODS: Precision was evaluated using 3-level control materials. Comparison analysis was performed on 200 samples: 100 normal and 100 abnormal samples. We evaluated the 5 part differential count, LIC, IMG, IMM, IML, ALY, and ERB. These parameters were analyzed in comparison with the results from the reference method, manual differential count. RESULTS: The coefficients of variation (CVs) of precision were <5% for neutrophils and lymphocytes, <15% for eosinophils and basophils and <7% for erythroblasts. The correlation coefficients (r) were >0.9 except monocytes and basophils. IMG, ALY and erythroblasts were also well correlated with manual count (r=0.8315, 0.5602, 0.8144, respectively). The efficiency of flagging system was 84% for LIC, 80% for ALY, and 78.0% for increased ERB (>2/100WBCs). CONCLUSIONS: The Pentra DX 120 performed reliable differential leukocyte and IMG, ALY, and ERB results demonstrated comparable performance to manual count. And, the flagging system was efficient for detecting each abnormal cell population. We expect the Pentra DX 120 double matrix and erythroblast count can reduce microscopic review rate in routine laboratory and promote laboratory efficiency.
Basophils ; Eosinophils ; Erythroblasts ; Fluorescence ; Granulocytes ; Leukocytes ; Lymphocytes ; Monocytes ; Neutrophils

BACKGROUND: Although the bone marrow transplantation is now recognized as an effective therapy for some hematologic and malignant diseases, HLA-matched related donor is limited. Human umbilical cord blood contains sufficient numbers of hematopoietic stem cells to provide long-term engraftment in the unrelated recipient with several advantages. The number of hematopoietic stem cells is an important factor for the better result in transplantation. The purpose of this study is the development of technique for the collection, separation and cryopreservation of the cord blood mononuclear cells for transplantation. METHODS: Forty-four cord bloods were collected during an uneventful delivery from the Kyungpook National University Hospital and Hana Hospital, Taegu, Korea from May to June, 1997. The mononuclear cell separation with Ficoll-Hypaque(Lymphocyte separation medium, Bionetics, USA) or red cell sedimentation with 3% gelatin(Sigma, USA) or 6% hydroxyethyl starch(HES; Fresenius, Germany) was done and the numbers and viabilities were compared with hemocytometry(Technicon H-3 RTX System, Irland). The viabilities of separated cells by different cell separators in the L-15 medium(Gifco, USA) containing 2% FBS(Gifco, USA) according to time were compared. The viabilities of separated cells by different cell separators in the L-15 medium(Gifco, USA) containing 17% FBS according to time and temperature were compared. The cell viabilities according to the different cell separating, freezing(direct, program-A or program-B) and thawing(rapid or slow) methods were compared. The method of direct freezing is putting the cells into the liquid nitrogen chamber(-196oC) directly. The method of program-A is lowering the temperature as 1oC/min to -90oC by programmed cell freezer(Cryomed 1010, USA) and then putting the cells into the liquid nitrogen chamber. The method of program-B is lowering the temperature as 1oC/min, to -4oC and 25oC/min, to -40oC, and raising as 15oC/min, to -12oC, and lowering again as 1oC/min, to -40oC and 10oC/min, to -90oC by programmed cell freezer and then putting the cells into the liquid nitrogen chamber. The method of rapid thawing is thawing the freezed cells in the 37oC incubator and the method of slow thawing is in the room temperature of 20oC. The functional aspect of stem cell after cryopreservation was performed with cell culture in the methylcellulose medium(Sigma, USA) and GM-CSF(LG Pharmacy, Korea) in the 5% CO2 incubator(37oC; Forma, USA). The statistical analysis was done with Analysis of Variance Procedure and T-test. RESULTS: The mean volume of cord blood was 121 mL(80~155 mL) and was no difference with maternal and fetal conditions, but the large volume could be obtained with rapid collection from early ligation of cord. The hemoglobin was 14.2+/-1.6 g/dL, hematocrit 46+/-6%, red blood cell 4.1+/-0.4x106/microL, platelet 236+/-82x103/microL and total white blood cell 11,893+/-4,010/microL with neutrophil 54.5+/-11.5%, eosinophil 2.6+/-1.3%, basophil 1.4+/-1.1%, lymphocyte 31.7+/-12.0% and monocyte 8.5+/-2.8%. The removal of red blood cell was 98.3% with F-H, 80.7% with gelatin and 54.3% with HES(P<0.0001). The removal of platelet was 64.8% with F-H, 85.2% with gelatin and 80.0% with HES. The removal of mononuclear cell was 73.3+/-15.4% with F-H, 99.1+/-22.4% with gelatin and 94.2+/-22.8% with HES(P<0.0001). The removal of polymorphonuclear cell was 91.0% with F-H, 42.3% with gelatin and 35.5% with HES(P<0.0001). The viabilities of separated cells by gelatin or HES in the L-15 medium containing 2% FBS for 18~24 hours were low compared to that by F-H and these differences were resolved with raising the concentration of FBS up to 17%. The program-B freezing(59.4%) was the best method in recovery of cells compared to the direct(36.9%) or program-A freezing(41.6%). But no differnces were observed according to the different cell separating or thawing methods. The colony formation from cryopreserved cell was observed at 7 th day of cell culture. CONCLUSION: In summary, the large volume of cord blood could be obtained with rapid collection after ligation of cord. The best way of mononuclear cell separation was the method with 3% gelatin among three the program-B freezing was the best way in recovery of cells after thawing these will be important factors to raise the success rate in hematopoietic stem cell transplantation.
Basophils ; Blood Platelets ; Bone Marrow Transplantation ; Cell Culture Techniques ; Cell Separation* ; Cell Survival ; Cryopreservation* ; Daegu ; Eosinophils ; Erythrocytes ; Fetal Blood* ; Freezing ; Gelatin ; Gyeongsangbuk-do ; Hematocrit ; Hematopoietic Stem Cell Transplantation ; Hematopoietic Stem Cells ; Humans* ; Incubators ; Korea ; Leukocytes ; Ligation ; Lymphocytes ; Methylcellulose ; Monocytes ; Neutrophils ; Nitrogen ; Pharmacy ; Stem Cells ; Tissue Donors

BACKGROUND: The XN-20 (Sysmex, Japan) is a recently developed hematology analyser, which adopts new technologies to improve the accuracy of complete blood count (CBC) and white blood cell (WBC) differentials and the efficiency of the flag system. In this study, we evaluated the performance of the XN-20 for CBC, WBC differentials, and reticulocyte counts. We also analysed the efficiency of its flag system. METHODS: We evaluated the precision and linearity of CBC and reticulocyte counts. In the correlation study, the results of XN-20 were compared with those obtained using ADVIA 2120 (Siemens, USA). The performance was also evaluated in the 'low WBC mode.' We analysed the efficiency of the flag system in detecting abnormal blood cells using 43 abnormal samples. RESULTS: The CVs for precision were <2% for most of the CBC parameters. Linearity was good for WBC, red blood cell (RBC), Hb, Hct, and platelet. The results of XN-20 were well correlated with those of ADVIA 2120. The correlation coefficients (r) was >0.9800 for all CBC parameters except for erythrocyte indices, and it was >0.9500 for WBC differentials except for monocyte and basophil. In the 'low WBC mode,' XN-20 could reliably analyse the WBC differentials in samples with low WBC count. The efficiencies of the flag systems were 95.3% for blasts, 83.7% for left-shifted neutrophils, 97.7% for atypical lymphocytes, and 86.0% for nucleated RBCs. CONCLUSIONS: The XN-20 showed good precision and its results were well correlated with those obtained using ADVIA 2120. In particular, in the 'low WBC mode,' it could provide reliable WBC differentials for samples with low WBC counts, and the flag systems detected abnormal blood cells with high efficiency.
Basophils ; Blood Cell Count* ; Blood Cells ; Blood Platelets ; Erythrocyte Indices ; Erythrocytes ; Hematology ; Leukocyte Count ; Leukocytes ; Lymphocytes ; Monocytes ; Neutrophils ; Reticulocyte Count ; Statistics as Topic

BACKGROUND: The XN-20 (Sysmex, Japan) is a recently developed hematology analyser, which adopts new technologies to improve the accuracy of complete blood count (CBC) and white blood cell (WBC) differentials and the efficiency of the flag system. In this study, we evaluated the performance of the XN-20 for CBC, WBC differentials, and reticulocyte counts. We also analysed the efficiency of its flag system. METHODS: We evaluated the precision and linearity of CBC and reticulocyte counts. In the correlation study, the results of XN-20 were compared with those obtained using ADVIA 2120 (Siemens, USA). The performance was also evaluated in the 'low WBC mode.' We analysed the efficiency of the flag system in detecting abnormal blood cells using 43 abnormal samples. RESULTS: The CVs for precision were <2% for most of the CBC parameters. Linearity was good for WBC, red blood cell (RBC), Hb, Hct, and platelet. The results of XN-20 were well correlated with those of ADVIA 2120. The correlation coefficients (r) was >0.9800 for all CBC parameters except for erythrocyte indices, and it was >0.9500 for WBC differentials except for monocyte and basophil. In the 'low WBC mode,' XN-20 could reliably analyse the WBC differentials in samples with low WBC count. The efficiencies of the flag systems were 95.3% for blasts, 83.7% for left-shifted neutrophils, 97.7% for atypical lymphocytes, and 86.0% for nucleated RBCs. CONCLUSIONS: The XN-20 showed good precision and its results were well correlated with those obtained using ADVIA 2120. In particular, in the 'low WBC mode,' it could provide reliable WBC differentials for samples with low WBC counts, and the flag systems detected abnormal blood cells with high efficiency.
Basophils ; Blood Cell Count* ; Blood Cells ; Blood Platelets ; Erythrocyte Indices ; Erythrocytes ; Hematology ; Leukocyte Count ; Leukocytes ; Lymphocytes ; Monocytes ; Neutrophils ; Reticulocyte Count ; Statistics as Topic

BACKGROUND: The BC-6800 (Mindray, China) is a recently developed hematology analyzer that utilizes 'SF Cube Technology' to improve the reliability of complete blood counts (CBC), white blood cell (WBC) differentials, and erythroblast counts. In this study, we evaluated the performance of the BC-6800 for CBC, WBC differentials, reticulocyte counts, and erythroblast counts and analyzed the efficiency of its flag system. METHODS: Specimens from 100 healthy controls and 95 patients were used. We performed precision and correlation studies of CBC, WBC differentials, reticulocyte counts, and erythroblast counts. We also analyzed the efficiency of the flag system in detecting abnormal blood cells. RESULTS: The coefficients of variation (CVs) of precision were <2% for most CBC parameters and <5% for neutrophil, eosinophil, and reticulocyte counts. The results obtained using the BC-6800 were well correlated with those of the ADVIA 2120 (Siemens, USA) and LH 750 (Beckman Coulter Corporation, USA). The correlation coefficients (r) were >0.9800 for CBC except erythrocyte indices, and >0.9500 for WBC differentials except monocyte and basophil. The WBC differentials and erythroblast counts obtained using the BC-6800 were well correlated with those of manual counts. The efficiencies of the flag system were 77.9% for Blasts, 82.1% for Immature Gran, 86.3% for Atypical Lymph, and 92.6% for NRBC present. CONCLUSIONS: The BC-6800 showed good precision and correlation with pre-existing hematology analyzers. The flag systems were quite efficient for detecting abnormal blood cells. Our study demonstrated that the BC-6800 hematology analyzer exhibits suitable performance and is helpful in routine laboratories.
Basophils ; Blood Cell Count ; Blood Cells ; Eosinophils ; Erythroblasts ; Erythrocyte Indices ; Hematology ; Humans ; Leukocytes ; Monocytes ; Neutrophils ; Reticulocyte Count ; Statistics as Topic

BACKGROUND: We evaluated the performance of leukocyte differential counting and clinical usefulness of the morphologic flags of the SE-000, and set optimal criteria for selecting and reviewing the specimens with increased abnormal cells. METHODS: From the results of SE-000 and manual leukocyte differential counting in 100 healthy control and 520 patient specimens we evaluated the correlations on the leukocyte fractions as well as the frequency, sensitivity and false positivity of the flags. After determination of the review criteria we calculated total review rate from the 3,403 consecutive CBC specimens. RESULTS: In both control and patient groups the correlation between two methods was high with the exception of monocytes and basophils. Regarding the morphologic flags, Blast was sensitive (86.9%) however could not detect mature looking lymphoblasts. Immature granulocyte showed high sensitivity (93.7%). Left shift showed the highest frequency (34.6%) and false positive rate (82.8%). Atypical lymphocytes and NRBC showed relatively low sensitivity (63.6%, and 50.5%, respectively). We determined to review the slide when 1) All morphologic flags except Left shift are marked, 2) WBC <3,000/microliter or >20,000/microliter, Hb <8.0 g/dL or 18.0 g/dL, Platelet <100,000/microliter or >600,000/microliter, 3) Severe deviation of leukocyte fractions or 4) Specially requested by physician. As a result, total review rate was 25.0% while 14 abnormal cases with no flags could be additionally detected. CONCLUSIONS: A new review criteria determined from the results of CBC and leukocyte differential together with morphologic flags could reduce the review rate without skipping the abnormal cases.
Basophils ; Blood Platelets ; Granulocytes ; Humans ; Leukocytes ; Lymphocytes ; Monocytes

BACKGROUND: We evaluated the performance of leukocyte differential counting and clinical usefulness of the morphologic flags of the SE-000, and set optimal criteria for selecting and reviewing the specimens with increased abnormal cells. METHODS: From the results of SE-000 and manual leukocyte differential counting in 100 healthy control and 520 patient specimens we evaluated the correlations on the leukocyte fractions as well as the frequency, sensitivity and false positivity of the flags. After determination of the review criteria we calculated total review rate from the 3,403 consecutive CBC specimens. RESULTS: In both control and patient groups the correlation between two methods was high with the exception of monocytes and basophils. Regarding the morphologic flags, Blast was sensitive (86.9%) however could not detect mature looking lymphoblasts. Immature granulocyte showed high sensitivity (93.7%). Left shift showed the highest frequency (34.6%) and false positive rate (82.8%). Atypical lymphocytes and NRBC showed relatively low sensitivity (63.6%, and 50.5%, respectively). We determined to review the slide when 1) All morphologic flags except Left shift are marked, 2) WBC <3,000/microliter or >20,000/microliter, Hb <8.0 g/dL or 18.0 g/dL, Platelet <100,000/microliter or >600,000/microliter, 3) Severe deviation of leukocyte fractions or 4) Specially requested by physician. As a result, total review rate was 25.0% while 14 abnormal cases with no flags could be additionally detected. CONCLUSIONS: A new review criteria determined from the results of CBC and leukocyte differential together with morphologic flags could reduce the review rate without skipping the abnormal cases.
Basophils ; Blood Platelets ; Granulocytes ; Humans ; Leukocytes ; Lymphocytes ; Monocytes